活性GLP-1（7-36）特異性酶免試劑盒
美國(guó)*
【型號(hào)】KT-871
【預(yù)期用途】
該高靈敏度ELISA（酶聯(lián)免疫吸附測(cè)定）試劑盒是專門用來(lái)定量檢測(cè)血清樣本中的胰高血糖素樣肽-1（7-36）[GLP-1（7-36）]的濃度。哺乳動(dòng)物中的GLP-1肽鏈的基本氨基酸序列*相同，如：小鼠、大鼠、豬、狗、猴、人等。該試劑盒僅適用于科學(xué)研究。
【實(shí)驗(yàn)原理】
這個(gè)ELISA的設(shè)計(jì)，研發(fā)和生產(chǎn)的目的是對(duì)血漿樣品中的活性GLP-1（7-36）進(jìn)行定量檢測(cè)。該試劑盒是基于運(yùn)用兩個(gè)GLP-1（7-36）特異性抗體分別與兩個(gè)位點(diǎn)結(jié)合的“雙位點(diǎn)夾心”技術(shù)。
將標(biāo)準(zhǔn)品、質(zhì)控品、和待測(cè)樣品加入鏈霉親和素包被的微孔板中。隨后，將生物素偶聯(lián)的的GLP-1（7-36）特異抗體和HRP標(biāo)記的GLP-1（7-36）特異抗體混合物添加到各個(gè)孔中。經(jīng)過(guò)*個(gè)保溫孵育期后，形成了“鏈霉親和素-生物素抗體- GLP-1（7-36）-HRP標(biāo)記抗體”的“雙位點(diǎn)夾心”結(jié)構(gòu)，該復(fù)合體被微孔板的內(nèi)壁吸附住。游離的HRP標(biāo)記抗體和緩沖基質(zhì)則在洗滌過(guò)程中被沖走。微孔板加入過(guò)氧化物酶基質(zhì)（3,3',5,5'-四甲基聯(lián)苯胺，TMB）并經(jīng)過(guò)一段時(shí)間的反應(yīng)后，用酶標(biāo)儀測(cè)量吸光度。微孔板內(nèi)壁的免疫復(fù)合體的酶活性與樣本中的GLP-1（7-36）濃度成正比。
【組成成分】
1. 鏈霉親和素包被的微孔板：1塊×96個(gè)
2. GLP-1示蹤抗體：1瓶×0.6mL
3. GLP-1（7-36）捕捉抗體：1瓶×0.6mL
4. ELISA濃縮洗滌液，30X：1瓶×30mL
5. ELISA HRP基質(zhì)：1瓶×24mL
6. ELISA終止液：1瓶×12mL
7. GLP-1標(biāo)準(zhǔn)品：6瓶
8. GLP-1質(zhì)控品：2瓶
9. 示蹤抗體稀釋液:1瓶×12mL
簡(jiǎn)要實(shí)驗(yàn)步驟：
1. 在各孔中添加100µL的標(biāo)準(zhǔn)品、質(zhì)控品和病人樣本；
2. 在各孔中添加100µL抗體混合物；
3. 2-8℃靜置培養(yǎng)20-24小時(shí)；
4. 用稀釋洗滌緩沖液洗凈條帶；
5. 各孔中添加200 µL的TMB基質(zhì)；
6. 室溫下靜置20分鐘；
7. 添加50 µL終止液；
Active GLP-1 （7-36） Specific ELISA Kit
Catalog Number: KT-871
Storage：This test kit must be stored at 2 – 8°C upon receipt.
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
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