RU2003124736A - METHOD FOR DETERMINING THE CONCENTRATION OF THE MAIN PROTEIN OF HEAT SHOCK 70 KDA - Google Patents

METHOD FOR DETERMINING THE CONCENTRATION OF THE MAIN PROTEIN OF HEAT SHOCK 70 KDA Download PDF

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Publication number
RU2003124736A
RU2003124736A RU2003124736/15A RU2003124736A RU2003124736A RU 2003124736 A RU2003124736 A RU 2003124736A RU 2003124736/15 A RU2003124736/15 A RU 2003124736/15A RU 2003124736 A RU2003124736 A RU 2003124736A RU 2003124736 A RU2003124736 A RU 2003124736A
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RU
Russia
Prior art keywords
wells
hsp70
washing
incubation
enzyme
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RU2003124736/15A
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Russian (ru)
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RU2242764C1 (en
Inventor
Сергей Станиславович Новоселов (RU)
Сергей Станиславович Новоселов
Сергей Алексеевич Суржиков (RU)
Сергей Алексеевич Суржиков
Алексей Петрович Воронин (RU)
Алексей Петрович Воронин
Ирина Владимировна Гужова (RU)
Ирина Владимировна Гужова
Борис Александрович Маргулис (RU)
Борис Александрович Маргулис
Original Assignee
Институт цитологии Российской академии наук (статус государственного учреждени ) (RU)
Институт цитологии Российской академии наук (статус государственного учреждения)
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Priority to RU2003124736/15A priority Critical patent/RU2242764C1/en
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Publication of RU2242764C1 publication Critical patent/RU2242764C1/en
Publication of RU2003124736A publication Critical patent/RU2003124736A/en

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Claims (1)

Способ определения концентрации основного белка теплового шока 70 кДа (БТШ70), заключающийся в иммобилизации агента, взаимодействующего с БТШ70 на поверхности лунок 96-луночного планшета для иммуноферментного анализа, промывке лунок буфером, внесении в лунки исследуемого образца или стандарта и инкубации, промывке лунок, внесении в лунки поликлональных кроличьих антител к БТШ70 и инкубации, промывке лунок, внесении в лунки антител, взаимодействующих с кроличьими антителами, коньюгированных с ферментом и инкубации, промывке лунок, внесении в лунки субстрата фермента и хромогена, инкубации, спектрофотометрической оценке развившейся окраски и соотнесения ее с данными калибровки, отличающийся тем, что в качестве иммобилизованного агента, взаимодействующего с БТШ70, используют производное АТФ, ковалентно связанное в комплексе с ГМДА с яичным альбумином, к образцам и стандартам добавляют натриевую соль АТФ до 10 мМ, а также вносят в буфер ионы магния до конечной концентрации 10 мМ на всех этапах.The method of determining the concentration of the main heat shock protein of 70 kDa (HSP70), which consists in immobilizing an agent that interacts with HSP70 on the surface of the wells of a 96-well plate for enzyme-linked immunosorbent assay, washing the wells with buffer, introducing into the wells of the test sample or standard and incubation, washing the wells, making into the wells of polyclonal rabbit antibodies to HSP70 and incubation, washing the wells, introducing into the wells antibodies interacting with rabbit antibodies conjugated with the enzyme and incubating, washing the wells, introducing into wells of the substrate of the enzyme and chromogen, incubation, spectrophotometric evaluation of the developed color and its correlation with the calibration data, characterized in that as an immobilized agent interacting with HSP70, an ATP derivative covalently coupled in combination with HMDA with egg albumin to samples and standards is used ATP sodium salt is added up to 10 mM, and magnesium ions are added to the buffer to a final concentration of 10 mM at all stages.
RU2003124736/15A 2003-08-07 2003-08-07 Method for detecting concentration of the main thermal shock protein 70 kda RU2242764C1 (en)

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RU2003124736/15A RU2242764C1 (en) 2003-08-07 2003-08-07 Method for detecting concentration of the main thermal shock protein 70 kda

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RU2242764C1 RU2242764C1 (en) 2004-12-20
RU2003124736A true RU2003124736A (en) 2005-02-10

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
UA112515C2 (en) * 2009-06-03 2016-09-26 Дау Аґросаєнсиз Елелсі METHOD OF DETERMINING THE PRESENCE OF TWO OR MORE PROTEINS OF INTEREST, IN SAMPLES OF VEGETABLE ORIGIN

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