CN108051582B - Improved immunohistochemical repair liquid - Google Patents
Improved immunohistochemical repair liquid Download PDFInfo
- Publication number
- CN108051582B CN108051582B CN201711286273.7A CN201711286273A CN108051582B CN 108051582 B CN108051582 B CN 108051582B CN 201711286273 A CN201711286273 A CN 201711286273A CN 108051582 B CN108051582 B CN 108051582B
- Authority
- CN
- China
- Prior art keywords
- minutes
- repair
- immunohistochemical
- antigen
- repair liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to an improved immunohistochemical repair liquid, which comprises trisodium citrate, NP-40 and an emulsifier, is infiltrated by the improved immunohistochemical repair liquid, has a better repair effect, and can repair tissue sections under a smaller dosage.
Description
Technical Field
The invention relates to the field of immunology, in particular to an improved immunohistochemical repair liquid.
Background
An antigen is a substance that stimulates the body to produce a (specific) immune response and that binds to the immune response product antibody and sensitized lymphocytes in vitro and in vivo to produce an immune effect (specific reaction). The basic properties of the antigen are heterogeneity, macromolecule and specificity. Foreign substances are antigenic substances that enter the body tissue and must be different from the components of the cells of the body tissue. Antigens generally refer to foreign substances entering the body, such as bacteria, viruses, pollen, etc.; the antigen can also be substances among different species, such as horse serum entering the body of a rabbit, and a plurality of proteins in the horse serum become the antigen substances of the rabbit; the substances between allogenic bodies can also become antigens, such as blood type, transplantation immunity and the like; certain isolated components in the body may also become antigens, such as eye crystallin, sperm cells, thyroglobulin, etc., which are normally immobilized in a part of the body isolated from antibody-producing cells and thus do not cause the production of antibodies in the body. Immunohistochemistry is the most commonly used method in clinical pathological diagnosis, and the principle is that antigens are specifically combined with antibodies, and color developing agents for marking the antibodies are developed through chemical reaction to determine antigens (polypeptides and proteins) in tissue cells, and the antigens are subjected to positioning, qualitative and quantitative researches. The immunohistochemical experiment mainly uses two types of tissue specimens and cell specimens, the paraffin section is the most common and most basic preferred method for manufacturing the tissue specimens, and the manufacturing process of the paraffin section has certain influence on the exposure of antigens in tissues but can carry out antigen repair.
The paraffin section specimens are fixed by formaldehyde, so that intracellular antigens form aldehyde bonds and carboxymethyl bonds to block part of antigenic determinants, and meanwhile, cross-linking is carried out among proteins to conceal the antigenic determinants. Therefore, antigen retrieval is required first when immunohistochemical staining is performed. Antigen retrieval is most important in immunohistochemical staining, directly determining staining results as well as diagnostic results.
There are many current immunohistochemical repair liquids, such as immunohistochemical repair liquid of DAKO denmurak a/S, immunohistochemical repair liquid of antibody diagnostic reagent limited in shanghai long island, biotechnology limited in shanghai long island, citrate immunohistochemical repair liquid produced by new biotechnology development limited in fuzhou, and the like. The immunohistochemical repair liquid comprises the following components: citric acid, trisodium citrate, preservatives, and the like.
The immunohistochemical repair liquid mainly contains citric acid, is a common main solvent for clinical immunohistochemistry, and is only suitable for general antigen repair. Many clinical antigens are difficult to detect, and the conventional immunohistochemical repair liquid has poor repair effect and directly influences subsequent dyeing and diagnosis.
Disclosure of Invention
In order to solve the problems, the invention provides an improved immunohistochemical repair liquid. The improved immunohistochemical repair liquid has a good repair effect, and can play a good repair role on a plurality of antigens which are difficult to detect.
According to one aspect of the present invention, the present invention provides an improved immunohistochemical repair solution, comprising: trisodium citrate, NP-40 and emulsifier.
Further, the paint comprises the following components in parts by weight: 20-40 parts of trisodium citrate, 0.1-4 parts of NP-40 and 2-20 parts of emulsifier.
Further, the paint comprises the following components in parts by weight: 25-35 parts of trisodium citrate, 0.5-2 parts of NP-40 and 5-15 parts of emulsifier.
Further, the paint comprises the following components in parts by weight: 30 parts of trisodium citrate, 0.75 part of NP-40 and 7.5 parts of emulsifier.
Further, the emulsifier is tween-20 or glycerol.
Further, the preparation method of the immunohistochemical repair liquid comprises the following steps: by means of H20, dissolving trisodium citrate, and adding NP-40; after complete dissolution, the emulsifier is added.
Further, the emulsifier is tween-20 or glycerol.
Compared with the prior art, the invention has the following advantages:
1. the improved immunohistochemical repair liquid disclosed by the invention is soaked, has a good repair effect, and can repair tissue slices with a small dosage;
2. the improved immunohistochemical repair solution of the present invention is suitable for many antigens that are difficult to detect clinically, such as Fas, Bax, FVIII, &lTtTtransfer = L "&gTt L &lTt/T &gTt amnin, CoIV, Keratin, CEA, GFAP, etc.;
3. the tissue section repaired by the improved immunohistochemical repair liquid is easy to stain and has better staining effect;
the foregoing description is only an overview of the technical solutions of the present invention, and the embodiments of the present invention are described below in order to make the technical means of the present invention more clearly understood and to make the above and other objects, features, and advantages of the present invention more clearly understandable.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The improved immunohistochemical repair liquid provided in the embodiment comprises: trisodium citrate, NP-40 and emulsifier. The improved immunohistochemical repair liquid takes trisodium citrate as a main repair active ingredient, and takes NP-40, Tween-20 and H2O as a mixed solvent.
As described above, the emulsifier has an emulsifying action, can improve the compatibility of each substance in the repair liquid, has little influence on the active ingredient in the repair liquid, and does not have much influence on the repair of the tissue section. Wherein the emulsifier is tween-20 or glycerol.
Tween 20 is a surfactant, and is a kind of macromolecule, and the molecule has both hydrophilic part and lipophilic part. Therefore, the plant can be promoted to absorb macromolecules which cannot be dissolved in water, and the moisture can be helped to permeate a certain high-fat biological membrane.
Glycerol, namely glycerol, is a colorless, odorless and sweet viscous liquid, has high permeability, can be mixed and dissolved with water in any proportion, has great hygroscopicity, is used for quickly removing intracellular water, and plays a role in protecting antigens.
NP-40(Nonidet P40), translated in Chinese as ethylphenylpolyethylene glycol, is a mild nonionic detergent; NP-40 is a very mild nonionic detergent, can basically destroy cell membranes at a concentration of 1%, has a weak effect on nuclear membranes, and can obtain cytoplasmic protein by combining with a specific buffer solution. Has strong binding force with protein, is used for preventing the interaction between substance molecules and hydrophobic molecules and ensures the full dissolution and the stable structure of the protein. Especially for the solubilization of membrane proteins under non-denaturing conditions. NP-4 plays a role in displaying intracellular antigens and can improve the exposure degree of antigens which are difficult to detect.
H in the improved immunohistochemical repair fluids of the present invention2O can be double distilled water or triple distilled water; double distilled water is one kind of redistilled water, and is obtained by redistilling water subjected to primary distillation; the triple distilled water is water subjected to tertiary distillation.
Example 1
Each 1000m of L modified immunohistochemical repair solution comprises 30g of trisodium citrate, 0.75m of L NP-40, 7.5m of L Tween-20 and the balance of double distilled water.
The preparation method comprises the following steps: weighing trisodium citrate, and dissolving with double distilled water; adding NP-40, adding Tween-20 after completely dissolving, and fixing the volume to the rated capacity by using double distilled water after the Tween-20 is dissolved.
Example 2
Each 1000m of L modified immunohistochemical repair solution comprises 20g of trisodium citrate, 4m of L NP-40, 2m of L glycerol and the balance of double distilled water.
The preparation method comprises the following steps: weighing trisodium citrate, and dissolving with triple distilled water; adding NP-40, adding glycerol after complete dissolution, and fixing the volume to the rated volume by double distilled water after the glycerol is dissolved.
Example 3
Each 1000m of L modified immunohistochemical repair solution comprises trisodium citrate 40g, NP-40 with 0.1m L, Tween-20 with 20m L and double distilled water in balance.
The preparation method is the same as that of example 1
Example 4
Every 1000m L modified immunohistochemical repair solution comprises 25g trisodium citrate, 2m L NP-40, 5m L glycerol and the balance of double distilled water.
The preparation method is the same as that of example 2
Example 5
Each 1000m of L modified immunohistochemical repair solution comprises 35g of trisodium citrate, 0.5m of L NP-40, 15m of L Tween-20 and the balance of double distilled water.
The preparation method is the same as that of example 1
Example 6
Each 1000m of L modified immunohistochemical repair solution comprises 23g of trisodium citrate, 3m L NP-40, 4m L Tween-20 and the balance of double distilled water.
The preparation method is the same as that of example 1
Example 7
Each 1000m of L modified immunohistochemical repair solution comprises trisodium citrate 38g, NP-40 0.2m L, glycerol 17m L and the balance of double distilled water.
The preparation method is the same as that of example 2
Test example 1 antigen retrieval of tissue sections containing GEA antigen
Experiment A: use of the improved immunohistochemical repair fluids of the present invention
1) Dewaxing the tissue slices in xylene for 5 minutes, and then sequentially putting the slices in 100% by volume alcohol for 2 minutes, 90% by volume alcohol for 2 minutes, 80% by volume alcohol for 2 minutes and 70% by volume alcohol for 2 minutes, and flushing the slices with Buffer (0.025 mol/L trisodium citrate) for 5 minutes;
2) heating the improved immunohistochemical repair liquid of example 1 to 92 ℃ at high temperature, placing the section in the repair liquid, covering a cover to start timing, and naturally cooling for 30 minutes at room temperature after 2 minutes; buffer rinse for 5 minutes;
3) peroxidase blocker for 5 minutes; buffer rinse for 5 minutes;
4) primary antibody (commercial murine anti-human monoclonal antibody) was incubated for 20 min; buffer rinse for 5 minutes;
5) secondary antibody (commercial enzyme-labeled goat anti-mouse IgG polymer) was incubated for 20 min; buffer rinse for 5 minutes;
6) performing DAB color development for 10 minutes; washing with tap water;
7) counterstaining with hematoxylin for 5 minutes; washing with tap water;
8) putting the slices into 70% volume ratio alcohol for 2 minutes, 80% volume ratio alcohol for 2 minutes, 90% volume ratio alcohol for 2 minutes and 100% volume ratio alcohol for 2 minutes respectively in sequence; xylene was clear for 5 minutes and mounted.
Experiment B: using other antigen retrieval solutions not according to the invention
The antigen retrieval fluid is selected from: citrate antigen retrieval solution produced by Fuzhou Mi New Biotechnology development Co.
The test steps are as follows:
a) the paraffin sections are placed in fresh dimethylbenzene to be soaked for 15 minutes and × 2 times, after excessive liquid is removed, the paraffin sections are placed in absolute ethyl alcohol to be soaked for 3 minutes and × 2 times, after excessive liquid is removed, the paraffin sections are placed in 95% ethyl alcohol to be soaked for 3 minutes, after excessive liquid is removed, the paraffin sections are placed in 85% ethyl alcohol to be soaked for 3 minutes, tap water is used for washing for 1 minute, and PBS solution is used for washing for 3 minutes and × 3 times.
b) The method comprises the steps of opening a pressure cooker, heating citric acid tissue antigen repair liquid on an induction cooker in a high-power mode until the citric acid tissue antigen repair liquid is boiled, placing slices subjected to dewaxing and hydration on a high-temperature-resistant dyeing rack, putting the slices into the pressure cooker, covering a cooker cover, fastening a pressure valve, continuing heating until steam is sprayed, keeping the time for 2 minutes, then leaving the pressure cooker from a heat source, naturally cooling for 5 minutes, showering the slices by tap water for cooling, removing the valve and opening the cover, taking out the slices after liquid in the cooker is naturally cooled to room temperature, washing for 3 minutes × 2 times by distilled water, and washing for 3 minutes × 3 times by PBS solution.
c) The PBS solution was removed and 100. mu.l of endogenous peroxidase blocker was added to the area of the tissue to be tested delineated by the oil pen and incubated for 10 minutes at room temperature and washed 3 minutes × 3 times with PBS solution.
d) The PBS solution was removed, 100. mu.l of primary or blank control reagent was added, incubated at room temperature for 60 minutes, and washed 3 minutes with PBS solution × 3 times.
e) The PBS solution was removed, 100. mu.l of enzyme-labeled goat anti-mouse IgG polymer was added, and the mixture was incubated at room temperature for 15 minutes, and washed with PBS solution for 3 minutes × 3 times.
f) And removing the PBS solution, adding 100-200 mul of freshly prepared DAB color development solution, incubating for 3-5 minutes, and observing the dyeing result by using a light microscope for generally not more than 10 minutes.
g) Washing with tap water, and adding 100-200 mul hematoxylin body cell staining solution for incubation for 10-30 seconds. The PBS solution or tap water is washed back to blue.
Experimental conclusion of test example 1: compared with the two methods, the section repaired by the experiment A has definite GEA positive positioning, clean background, dark brown color after dyeing, good contrast and little possibility of flaking off; in the section repaired by the experiment B, the GEA positive is basically positioned, but the section is light brown after being dyed, and the section stripping rate is higher, so that the effect of the improved immunohistochemical repair liquid is better than that of the antigen repair liquid in the experiment B in the antigen repair contrast experiment of the tissue section containing the GEA antigen.
Test example 2 tissue section antigen repair containing GFAP antigen
Experiment C: use of the improved immunohistochemical repair fluids of the present invention
1) Dewaxing the tissue slices in xylene for 5 minutes; gradient of 100% alcohol, 90% alcohol, 80% alcohol, 70% alcohol each for 2 minutes; buffer rinse for 5 minutes;
2) firstly, heating the improved immunohistochemical repair liquid to 92 ℃ at high temperature, placing the slices in the repair liquid, covering a cover, heating until the time of gas evolution begins to be timed for 2 minutes, and naturally cooling for 30 minutes; buffer rinse for 5 minutes;
3) peroxidase blocker for 5 minutes; buffer rinse for 5 minutes;
4) primary antibody (commercial murine anti-human monoclonal antibody) was incubated for 20 min; buffer rinse for 5 minutes;
5) secondary antibody (commercial enzyme-labeled goat anti-mouse IgG polymer) was incubated for 20 min; buffer rinse for 5 minutes;
6) performing DAB color development for 10 minutes; washing with tap water;
7) hematoxylin counterstain for 5 minutes; washing with tap water;
8) putting the slices into 70% alcohol, 80% alcohol, 90% alcohol and 100% alcohol in sequence, and performing gradient treatment for 2 minutes respectively; xylene was clear for 5 minutes and mounted.
Experiment D: using other antigen retrieval solutions not according to the invention
The antigen retrieval fluid is selected from: citrate antigen retrieval solution produced by Fuzhou Mi New Biotechnology development Co.
The test steps are as follows:
a) the paraffin sections are placed in fresh dimethylbenzene to be soaked for 15 minutes and × 2 times, after excessive liquid is removed, the paraffin sections are placed in absolute ethyl alcohol to be soaked for 3 minutes and × 2 times, after excessive liquid is removed, the paraffin sections are placed in 95% ethyl alcohol to be soaked for 3 minutes, after excessive liquid is removed, the paraffin sections are placed in 85% ethyl alcohol to be soaked for 3 minutes, tap water is used for washing for 1 minute, and PBS solution is used for washing for 3 minutes and × 3 times.
b) The method comprises the steps of opening a pressure cooker, heating citric acid tissue antigen repair liquid on an induction cooker in a high-power mode until the citric acid tissue antigen repair liquid is boiled, placing slices subjected to dewaxing and hydration on a high-temperature-resistant dyeing rack, putting the slices into the pressure cooker, covering a cooker cover, fastening a pressure valve, continuing heating until steam is sprayed, keeping the time for 2 minutes, then leaving the pressure cooker from a heat source, naturally cooling for 5 minutes, showering the slices by tap water for cooling, removing the valve and opening the cover, taking out the slices after liquid in the cooker is naturally cooled to room temperature, washing for 3 minutes × 2 times by distilled water, and washing for 3 minutes × 3 times by PBS solution.
c) The PBS solution was removed and 100. mu.l of endogenous peroxidase blocker was added to the area of the tissue to be tested delineated by the oil pen and incubated for 10 minutes at room temperature and washed 3 minutes × 3 times with PBS solution.
d) The PBS solution was removed, 100. mu.l of primary or blank control reagent was added, incubated at room temperature for 60 minutes, and washed 3 minutes with PBS solution × 3 times.
e) The PBS solution was removed, 100. mu.l of enzyme-labeled goat anti-mouse IgG polymer was added, and the mixture was incubated at room temperature for 15 minutes, and washed with PBS solution for 3 minutes × 3 times.
f) And removing the PBS solution, adding 100-200 mul of freshly prepared DAB color development solution, incubating for 3-5 minutes, and observing the dyeing result by using a light microscope for generally not more than 10 minutes.
g) Washing with tap water, and adding 100-200 mul hematoxylin body cell staining solution for incubation for 10-30 seconds. The PBS solution or tap water is washed back to blue.
Experimental conclusion of experimental example 2: compared with the two methods, the slice repaired in the experiment A has definite GFAP positive positioning, clean background, dark brown color after dyeing, good contrast and little possibility of stripping; the slice repaired by the experiment B has unclear GFAP positive positioning, more non-specific coloring and fuzzy staining and slight flaking, so in the contrast experiment of the tissue slice antigen repair containing GFAP antigen, the effect of the improved immunohistochemical repair liquid is better than that of the antigen repair liquid in the experiment D.
Test example 3 component screening of the improved immunohistochemical repair solution of the present invention
TABLE 1 amounts of the respective components added in the test examples
Tests 3-1 to 3-5 the 5 improved immunohistochemical repair solutions were prepared according to the method described in example 1, and the 5 improved immunohistochemical repair solutions were tested according to test example 1, test a.
Experimental results of experiment 3: tests 3-1 to 3-5 all achieve better positive positioning, and test 3-1 is light brown after dyeing and has a stripping sheet; test 3-2 was stained to light brown with flaking; experiment 3-3 was dark brown with clear background and no flaking after dyeing; tests 3-4 were dark brown in color with a clear background and slight flaking off; the test 3-5 is light brown after dyeing, the background is occasionally interfered with color, and the test has no flaking; therefore, in the comparative experiment of the antigen retrieval of tissue sections containing GFAP antigen, the effect of the modified immunohistochemical retrieval solution of experiment 3-3 was superior to that of the modified immunohistochemical retrieval solution of other groups.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (2)
1. An improved immunohistochemical repair liquid for GEA antigen or GFAP antigen is characterized by consisting of trisodium citrate, NP-40, an emulsifier and water, wherein every 1000m L of the immunohistochemical repair liquid consists of 30g of trisodium citrate, 0.75m L NP-40, 7.5m L Tween-20 and the balance of double distilled water.
2. The improved immunohistochemical repair solution for GEA antigen or GFAP antigen according to claim 1, wherein the preparation method of the immunohistochemical repair solution comprises the steps of:
by means of H2Dissolving trisodium citrate by using O, and adding NP-40; after complete dissolution, tween-20 was added.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711286273.7A CN108051582B (en) | 2017-12-07 | 2017-12-07 | Improved immunohistochemical repair liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711286273.7A CN108051582B (en) | 2017-12-07 | 2017-12-07 | Improved immunohistochemical repair liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108051582A CN108051582A (en) | 2018-05-18 |
| CN108051582B true CN108051582B (en) | 2020-08-04 |
Family
ID=62123161
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711286273.7A Active CN108051582B (en) | 2017-12-07 | 2017-12-07 | Improved immunohistochemical repair liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108051582B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110346552A (en) * | 2019-07-25 | 2019-10-18 | 深圳褀氏生物医疗电子有限公司 | A kind of universal antigen retrieval buffer |
| CN113495138A (en) * | 2021-06-22 | 2021-10-12 | 南京弗瑞思生物科技有限公司 | Tissue slice antigen repair liquid and use method thereof |
| CN113686634A (en) * | 2021-08-10 | 2021-11-23 | 杭州迪英加科技有限公司 | Dewaxing repair liquid beneficial to acquisition of paraffin section images |
| CN113740522B (en) * | 2021-09-01 | 2023-05-12 | 陕西脉元生物科技有限公司 | Antigen retrieval liquid, cell climbing tablet prepared from same, preparation method and application of cell climbing tablet |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7867443B2 (en) * | 2004-07-23 | 2011-01-11 | Dako Denmark A/S | Method and apparatus for automated pre-treatment and processing of biological samples |
| CN105987839B (en) * | 2016-06-03 | 2018-09-28 | 浙江世纪康大医疗科技股份有限公司 | A kind of EDTA- citric acid antigens reparation liquid |
-
2017
- 2017-12-07 CN CN201711286273.7A patent/CN108051582B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108051582A (en) | 2018-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108051582B (en) | Improved immunohistochemical repair liquid | |
| Foster et al. | Monoclonal antibodies to the human mammary gland: I. Distribution of determinants in non-neoplastic mammary and extra mammary tissues | |
| Vielkind et al. | A simple fixation procedure for immunofluorescent detection of different cytoskeletal components within the same cell | |
| CN101768218B (en) | Preparation method for PCV-II Cap protein monoclonal antibody, antibody and application | |
| Sandilands et al. | Lymphocytes with T and B cell properties in a lymphoproliferative disorder | |
| Georgala et al. | A rapid immunofluorescence technique for detecting salmonellae in raw meat | |
| Diamond et al. | The importance of Rh inhibitor substance in anti-Rh serums | |
| Takiya et al. | Connective matrix organization in human pulmonary fibrosis: collagen polymorphism analysis in fibrotic deposits by immunohistological methods | |
| Garrett et al. | Fibroblast-like cells from embryonic chick cornea, heart, and skin are antigenically distinct | |
| CN104237504A (en) | Immunofluorescence method and kit for pathological autopsy of anaphylactic shock | |
| Belák et al. | Comparative studies on the pathogenicity and tissue distribution of three virulence variants of classical swine fever virus, two field isolates and one vaccine strain, with special regard to immunohistochemical investigations | |
| CN104596827A (en) | Kit and staining method for doubly staining mycobacterium tuberculosis and antigen of mycobacterium tuberculosis | |
| JP3723204B1 (en) | Impermeable tissue quick fixative | |
| CN105548569A (en) | Detection method for peripheral blood VEGF of renal cancer patient | |
| Jafari et al. | Diagnostic value of immunoperoxidase staining and immunofluorescence in the study of kidney biopsy specimens | |
| Kilic et al. | Methods to evaluate the formation and stabilization of blood vessels and their role in tumor growth and metastasis | |
| Kaplan et al. | Immunologic Studies of Heart Tissue: II. Differentiation of a Myocardial Sarcoplasmic Antigen and Cardiolipin | |
| CN105585627A (en) | Antigen polypeptide and application of anti-GBM nephritis model built from same | |
| Emmart et al. | The localization of glyceraldehyde-3-phosphate dehydrogenase within the muscle of the roach, Periplaneta americana, by means of fluorescent antibody | |
| Jeurissen et al. | Immunocytochemical techniques to investigate the pathogenesis of infectious micro-organisms and the concurrent immune response of the host | |
| SE468231B (en) | PROCEDURE TO PROVIDE TASTY LUNGCARCINOMANTIGEN AND USE OF ANTIBODIES AGAINST FUCOSYLSYLOSYLGYLIOTETRAOS FOR PREPARATION OF COMPOSITION FOR TREATMENT OR VIVO / DIAGNOSIS | |
| Cave et al. | A cytochemical and autoradiographic investigation of nucleolar phospholipids | |
| Gray et al. | Immunofluorescence identification of Thermopolyspora polyspora, the causative agent of farmer's lung | |
| Knop et al. | Anti-P. acnes antibody in comedonal extracts | |
| Kåresen et al. | Induction of thyroiditis in guinea-pigs by intravenous injection of rabbit anti-guinea-pig thyroglobulin serum: II. Studies with fluorescent antibody technique |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PP01 | Preservation of patent right |
Effective date of registration: 20220430 Granted publication date: 20200804 |
|
| PP01 | Preservation of patent right |