嘉興行健生物科技有限公司
主營產(chǎn)品: 人穩(wěn)定型骨鈣素(1-43/49)ELIsA試劑盒,人酶免試劑盒,大小鼠酶免試劑盒,猴酶免試劑盒,牛酶免試劑盒 |
嘉興行健生物科技有限公司
主營產(chǎn)品: 人穩(wěn)定型骨鈣素(1-43/49)ELIsA試劑盒,人酶免試劑盒,大小鼠酶免試劑盒,猴酶免試劑盒,牛酶免試劑盒 |
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總GLP-1酶免試劑盒
美國*
【型號(hào)】 KT-876
【預(yù)期用途】
該高靈敏度ELISA(酶聯(lián)免疫吸附測定)試劑盒是專門用來定量檢測血漿樣本中的胰高血糖素樣肽-1(7-36)[GLP-1(7-36)]和(9-36)[GLP-1(9-36)]的總濃度。哺乳動(dòng)物的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】
該ELISA的設(shè)計(jì),研發(fā)和生產(chǎn)的目的是對血清樣品中的活性GLP-1(7-36)和GLP-1(9-36)進(jìn)行定量檢測。它是基于運(yùn)用兩個(gè)GLP-1特異性抗體與兩個(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GLP-1特異抗體和HRP標(biāo)記的GLP-1特異抗體混合物添加到各個(gè)孔中。經(jīng)過*個(gè)保溫孵育期,形成了“鏈霉親和素-生物素抗體- GLP-1(7-36)/(9-36)-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)則在洗滌過程中被沖走。微孔板加入過氧化物酶基質(zhì)(3,3',5,5'-四甲基聯(lián)苯胺,TMB)并經(jīng)過一段時(shí)間的反應(yīng)后,然后用酶標(biāo)儀測量吸光度。微孔板內(nèi)壁的免疫復(fù)合體與GLP-1(7-36)/(9-36)的連接酶活性與樣本中的總GLP-1數(shù)量成正比。
【組成成分】
1. 鏈霉親和素包被的微孔板:1塊×96個(gè)
2. 總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×20mL
5. ELISA HRP基質(zhì):1瓶×24mL
6. ELISA終止液:1瓶×12mL
7. 總GLP-1標(biāo)準(zhǔn)品:5瓶
8. GLP-1質(zhì)控品:2瓶
9. 示蹤抗體稀釋液:1瓶×12mL
【簡要實(shí)驗(yàn)步驟】
1. 在各孔中分別添加100µL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. 用洗滌緩沖液稀釋液洗凈條帶;
5. 在各孔中添加200 µL的TMB基質(zhì);
6. 室溫下靜置20分鐘;
7. 添加50 µL終止液;
8. 讀取450nm/620nm和450nm/650nm下的吸光度。
Total GLP-1 ELISA Kit
Catalog Number: KT-876
This ELISA (enzyme-linked immunosorbent assay) kit is produced for the quantitative determination of the total value of glucagon-like peptide-1 (7-36) [GLP-1 (7-36)] and (9-36) [GLP-1 (9-36)] 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.
This ELISA is designed, developed and produced for the quantitative measurement of GLP-1 (7-36) and (9-36) in plasma sample. The assay utilizes the two-site “sandwich” technique with two selected GLP-1 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 specific antibody and a horseradish peroxidate (HRP) conjugated GLP-1 specific antibody is added to each well. After the first incubation period, a “sandwich” immunocomplex of “Streptavidin – Biotin-Antibody – GLP-1(7-36)/(9-36) – 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)/(9-36) on the wall of the microtiter well is directly proportional to the amount of Total GLP-1 in the sample.
REAGENTS: Preparation
1. Streptavidin Coated Microplate:96 wells
2. Total GLP-1 Tracer Antibody:1vial× 0.6 mL
3. Total GLP-1 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. Total GLP-1 Standards:5 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