CN109100503A - 一种免疫组化笔液体及其制备方法 - Google Patents
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Abstract
本发明涉及一种免疫组化笔液体及其制备方法,该免疫组化笔液体包括以下重量百分比的组分:乙醇20%~30%;中性树胶15%~25%;丁酮20%~30%;环己酮15%~25%;丙酮15%~25%;聚二甲基硅氧烷1%~5%;气相二氧化硅0.2%~1.5%。制备时,将乙醇与中性树胶混合后取上清液,与环己烷、丁酮、二甲基硅氧烷混合,经气相二氧化硅充分分散后装瓶即得所述免疫组化笔液体。与现有技术相比,本发明可减少抗体和试剂用量,避免染色时液体扩散,提高操作速度。通过本配方制备的产品,其贴片和疏水更好、成本更低。
Description
技术领域
本发明涉及生物染色体技术领域,具体涉及一种免疫组化笔液体及其制备方法。
背景技术
免疫组化笔液体广泛应用于玻璃切片的各种免疫组织化学染色实验,如PAP法,ABC法,免疫荧光法,冰冻切片及原位杂交技术,可减少抗体和试剂用量,避免染色时液体扩散,提高操作速度。目前,商业化的免疫组化笔液体配方均为国外研制,国内主要为凡士林、石蜡等简单的混合液,至今尚未有相关专利申请。此外,目前商品化的免疫组化笔液贴片和疏水效果差,且价格昂贵。
发明内容
本发明的目的就是为了克服上述现有技术存在的缺陷而提供一种疏水性好、成本低的免疫组化笔液体及其制备方法。
本发明的目的可以通过以下技术方案来实现:一种免疫组化笔液体,该免疫组化笔液体包括以下重量百分比的组分:
其中聚二甲基硅氧烷具有良好疏水性以及较低表面能,用以在成型笔迹表面形成疏水层;疏水性气相二氧化硅能够进一步增加疏水性,并调节最终混合物的粘度;中性树胶用以与乙醇反应沉淀后,去除难溶成分,剩余粘性物质用以增加与载玻片玻璃表面的附着力,避免笔迹在免疫组化各种操作产生移位;丁酮、环己酮、丙酮为混合溶剂,用于溶解乙醇与中性树胶反应后成分,并将聚二甲基硅氧烷均匀分散形成胶体。
优选的,所述的中性树胶为纯度≥99%的HPLC。该中性树胶能更好地避免带入杂质。
优选的,所述的气相二氧化硅为疏水性气相二氧化硅,所述气相二氧化硅的比表面积>200m2/g。气相二氧化硅比表面积与粒径负相关,需要保证粒径足够小以避免阻塞组化笔纤维笔头。
优选的,所述的聚二甲基硅氧烷的粘度为500~1000cp,聚二甲基硅氧烷粘度与分子量正相关,适当分子量可以保证对表面活性剂的耐受性。
一种如上所述免疫组化笔液体的制备方法,包括以下步骤:
(1)将乙醇和中性树胶均匀混合,静置,取上清液;
(2)将步骤(1)所得上清液与丁酮、环己酮、丙酮、聚二甲基硅氧烷混合,然后加入气相二氧化硅,静置,即得所述免疫组化笔液体。
所述的乙醇和中性树胶均匀混合后的静置时间为12~24h。
所述的气相二氧化硅加入后静置时间为30~60min。
与现有技术相比,本发明的有益效果体现在以下几方面:
(1)该免疫组化笔液体的疏水性好,可以保证免疫组化效果;
(2)本发明采用的都是常用的物质,价格低廉,生产成本低。
具体实施方式
下面对本发明的实施例作详细说明,本实施例在以本发明技术方案为前提下进行实施,给出了详细的实施方式和具体的操作过程,但本发明的保护范围不限于下述的实施例。
实施例1
一种免疫组化笔(疏水画圈笔)液体配方,各组分及成分如表1所示。其中溶剂为丁酮-环己酮-丙酮-乙醇混合体系,疏水剂为聚二甲基硅氧烷,增稠剂为气相二氧化硅,其它助剂为中性树胶。
表1免疫组化笔液体配方
材料名称 | 样品1 | 样品2 | 样品3 |
乙醇(g) | 30 | 28 | 26 |
中性树胶(g) | 25 | 22 | 20 |
丁酮(g) | 30 | 28 | 25 |
环己酮(g) | 25 | 20 | 18 |
丙酮(g) | 25 | 20 | 18 |
聚二甲基硅氧烷(g) | 2.5 | 2.0 | 1.5 |
气相二氧化硅(g) | 0.5 | 1.0 | 1.5 |
制备时,先将乙醇与中性树胶(分析纯)1:1混合,摇匀,静置12小时后取上清液;然后将上述上清液与丁酮、环己酮、丙酮、二甲基硅氧烷混合,然后加入气相二氧化硅,装瓶。在使用时,(4)笔芯充入4ml,静置30分钟后即可使用。
样品1~3的产品经装笔实验后与目前市场上销售的笔比较,本发明制作的免疫组化笔在相同面积下可容纳更多的抗体稀释液,且不扩散,说明1~3的产品均可取得较好疏水效果。此外,本发明产品应用于免疫组织化学染色实验后,荧光染色更清晰。
实施例2
一种免疫组化笔(疏水画圈笔)液体配方,各组分及成分如表2所示。其中溶剂为丁酮-环己酮-丙酮-乙醇混合体系,疏水剂为聚二甲基硅氧烷,增稠剂为气相二氧化硅,其它助剂为中性树胶。
表2免疫组化笔液体配方
材料名称 | 样品4 | 样品5 | 样品6 | 样品7 | 样品8 |
乙醇(g) | 20 | 30 | 20 | 20 | 20 |
中性树胶(g) | 15 | 15 | 18.8 | 18.8 | 25 |
丁酮(g) | 30 | 20 | 20 | 20 | 20 |
环己酮(g) | 15 | 15 | 15 | 25 | 15 |
丙酮(g) | 15 | 15 | 25 | 15 | 17 |
聚二甲基硅氧烷(g) | 3.5 | 4.8 | 1 | 1 | 2 |
气相二氧化硅(g) | 1.5 | 0.2 | 0.2 | 0.2 | 2 |
制备时,先将乙醇与中性树胶(分析纯)1:1混合,摇匀,静置24小时后取上清液;然后将上述上清液与丁酮、环己酮、丙酮、二甲基硅氧烷混合,然后加入气相二氧化硅,装瓶。在使用时,(4)笔芯充入4ml,静置60分钟后即可使用。
样品4~8的产品经装笔实验后与目前市场上销售的笔比较,本发明制作的免疫组化笔在相同面积下可容纳更多的抗体稀释液,且不扩散,说明4~8的产品均可取得较好疏水效果。此外,本发明产品应用于免疫组织化学染色实验后,荧光染色更清晰。
Claims (7)
1.一种免疫组化笔液体,其特征在于,该免疫组化笔液体包括以下重量百分比的组分:
2.根据权利要求1所述的一种免疫组化笔液体,其特征在于,所述的中性树胶为纯度≥99%的HPLC。
3.根据权利要求1所述的一种免疫组化笔液体,其特征在于,所述的气相二氧化硅为疏水性气相二氧化硅,所述气相二氧化硅的比表面积>200m2/g。
4.根据权利要求1所述的一种免疫组化笔液体,其特征在于,所述的聚二甲基硅氧烷的粘度为500~1000cp。
5.一种如权利要求1~4任一所述免疫组化笔液体的制备方法,其特征在于,包括以下步骤:
(1)将乙醇和中性树胶均匀混合,静置,取上清液;
(2)将步骤(1)所得上清液与丁酮、环己酮、丙酮、聚二甲基硅氧烷混合,然后加入气相二氧化硅,静置,即得所述免疫组化笔液体。
6.根据权利要求5所述的一种免疫组化笔液体的制备方法,其特征在于,所述的乙醇和中性树胶均匀混合后的静置时间为12~24h。
7.根据权利要求5所述的一种免疫组化笔液体的制备方法,其特征在于,所述的气相二氧化硅加入后静置时间为30~60min。
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