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生产厂家厂商性质
嘉兴市所在地
活性GLP-1(7-36)特异性酶免试剂盒
美国*
【型号】KT-871
【预期用途】
该高灵敏度ELISA(酶联免疫吸附测定)试剂盒是专门用来定量检测血清样本中的胰高血糖素样肽-1(7-36)[GLP-1(7-36)]的浓度。哺乳动物中的GLP-1肽链的基本氨基酸序列*相同,如:小鼠、大鼠、猪、狗、猴、人等。该试剂盒仅适用于科学研究。
【实验原理】
这个ELISA的设计,研发和生产的目的是对血浆样品中的活性GLP-1(7-36)进行定量检测。该试剂盒是基于运用两个GLP-1(7-36)特异性抗体分别与两个位点结合的“双位点夹心”技术。
将标准品、质控品、和待测样品加入链霉亲和素包被的微孔板中。随后,将生物素偶联的的GLP-1(7-36)特异抗体和HRP标记的GLP-1(7-36)特异抗体混合物添加到各个孔中。经过*个保温孵育期后,形成了“链霉亲和素-生物素抗体- GLP-1(7-36)-HRP标记抗体”的“双位点夹心”结构,该复合体被微孔板的内壁吸附住。游离的HRP标记抗体和缓冲基质则在洗涤过程中被冲走。微孔板加入过氧化物酶基质(3,3',5,5'-四甲基联苯胺,TMB)并经过一段时间的反应后,用酶标仪测量吸光度。微孔板内壁的免疫复合体的酶活性与样本中的GLP-1(7-36)浓度成正比。
【组成成分】
1. 链霉亲和素包被的微孔板:1块×96个
2. GLP-1示踪抗体:1瓶×0.6mL
3. GLP-1(7-36)捕捉抗体:1瓶×0.6mL
4. ELISA浓缩洗涤液,30X:1瓶×30mL
5. ELISA HRP基质:1瓶×24mL
6. ELISA终止液:1瓶×12mL
7. GLP-1标准品:6瓶
8. GLP-1质控品:2瓶
9. 示踪抗体稀释液:1瓶×12mL
简要实验步骤:
1. 在各孔中添加100µL的标准品、质控品和病人样本;
2. 在各孔中添加100µL抗体混合物;
3. 2-8℃静置培养20-24小时;
4. 用稀释洗涤缓冲液洗净条带;
5. 各孔中添加200 µL的TMB基质;
6. 室温下静置20分钟;
7. 添加50 µL终止液;
Active GLP-1 (7-36) Specific ELISA Kit
Catalog Number: KT-871
INTENDED USE
This high sensitive ELISA (enzyme-linked immunosorbent assay) kit is produced for the exclusively quantitative determination of bioactive glucagon-like peptide-1 (7-36) [GLP-1 (7-36)] level in plasma samples. The primary amino acid sequence of GLP-1 peptide is identical among mammalian species, i.e. rat, mouse, pig, human, etc. This kit is for research purpose only.
ASSAY PRINCIPLE
This ELISA is designed, developed and produced for the quantitative measurement of bioactive GLP-1 (7-36) in plasma sample. The assay utilizes the two-site “sandwich” technique with two selected GLP-1 (7-36) specific antibodies.
Assay standards, controls and test samples are directly added to wells of a microplate that is coated with streptavidin. Subsequently, a mixture of biotinylated GLP-1 (7-36) specific antibody and a horseradish peroxidate (HRP) conjugated GLP-1 (7-36) specific antibody is added to each well. After the first incubation period, a “sandwich” immunocomplex of “Streptavidin – Biotin-Antibody – GLP-1(7-36) – HRP conjugated antibody” is formed and attached to the wall of the plate. The unbound HRP conjugated antibody is removed in a subsequent washing step. For the detection of this immunocomplex, each well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to GLP-1 (7-36) on the wall of the microtiter well is directly proportional to the amount of GLP-1 (7-36) in the sample.
REAGENTS: Preparation
1. Streptavidin Coated Microplate:96 wells
2. GLP-1 Tracer Antibody:1vial× 0.6 mL
3. GLP-1 (7-36) Capture Antibody:1vial× 0.6 mL
4. ELISA Wash Concentrate,30X:1 bottle× 20mL
5. ELISA HRP Substrate :1 bottle× 24mL
6. ELISA Stop Solution:1 bottle× 12mL
7. GLP-1 Standards:6 vials
8. GLP-1 Controls:2 vials
9. Tracer Antibody Diluent:1vials× 12mL
Short Assay Protocol:
- Add 100 μl/well of standards, control and patient sample
- Add 100 μl of Antibody Mixture
- Incubate 20 - 24 hour at 2-8°C, static
- Wash strips with diluted wash buffer
- Add 200 μl/well of TMB substrate
- Incubate 20 min at RT, static
- Add 50 μl stop solution
- Read strips at OD 450 nm/620 nm or 450 nm/650 nm