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上海恒遠(yuǎn)高品質(zhì)長期供應(yīng)“大鼠CD44分子ELISA試劑盒 "，產(chǎn)品種類齊全,、現(xiàn)貨供應(yīng),、質(zhì)量長期保障，提供完整的售前,，售中售后服務(wù),，歡迎各位新老客戶惠顧！
詳細(xì)介紹
產(chǎn)品名稱：大鼠CD44分子ELISA試劑盒
英文名稱：Rat cluster Of differentiation,CD44 ELISA KIT
大鼠CD44分子ELISA試劑盒操作步驟
1. 標(biāo)準(zhǔn)品的稀釋：本試劑盒提供原倍標(biāo)準(zhǔn)品一支,，用戶可按照下列圖表在小試管中進(jìn)行稀釋,。
| 2400 ng/L | 5號標(biāo)準(zhǔn)品 | 150μl的原倍標(biāo)準(zhǔn)品加入150μl標(biāo)準(zhǔn)品稀釋液 |
| 1200ng/L | 4號標(biāo)準(zhǔn)品 | 150μl的5號標(biāo)準(zhǔn)品加入150μl標(biāo)準(zhǔn)品稀釋液 |
| 600ng/L | 3號標(biāo)準(zhǔn)品 | 150μl的4號標(biāo)準(zhǔn)品加入150μl標(biāo)準(zhǔn)品稀釋液 |
| 300ng/L | 2號標(biāo)準(zhǔn)品 | 150μl的3號標(biāo)準(zhǔn)品加入150μl標(biāo)準(zhǔn)品稀釋液 |
| 150ng/L | 1號標(biāo)準(zhǔn)品 | 150μl的2號標(biāo)準(zhǔn)品加入150μl標(biāo)準(zhǔn)品稀釋液 |
3. 溫育：用封板膜封板后置37℃溫育30分鐘,。
4. 配液：將30倍濃縮洗滌液用蒸餾水30倍稀釋后備用
5. 洗滌：小心揭掉封板膜,，棄去液體，甩干,，每孔加滿洗滌液,，靜置30秒后棄去，如此重復(fù)5次,，拍干,。
6. 加酶：每孔加入酶標(biāo)試劑50μl，空白孔除外,。
7. 溫育：操作同3,。
8. 洗滌：操作同5。
9. 顯色：每孔先加入顯色劑A50μl,，再加入顯色劑B50μl,，輕輕震蕩混勻,，37℃避光顯色15分鐘.
10. 終止：每孔加終止液50μl，終止反應(yīng)（此時(shí)藍(lán)色立轉(zhuǎn)黃色）,。
11. 測定：以空白空調(diào)零,，450nm波長依序測量各孔的吸光度（OD值）。 測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行,。
Assay procedure
1. Dilute and add sample:Dilute Original density Standard as follow table:
| 800ng/ml | 5 Standard | 150μl Original density Standard+150μl Standard diluent |
| 400ng/ml | 4 Standard | 150μl 5 Standard+150μl Standard diluent |
| 200ng/ml | 3 Standard | 150μl 4 Standard+150μl Standard diluent |
| 100ng/ml | 2 Standard | 150μl 3 Standard +150μl Standard diluent |
| 50ng/ml | 1 Standard | 150μl 2 Standard +150μl Standard diluent |
2.add sample：Set blank wells separay （blank comparison wells don’t add sample and HRP-Conjugate reagent, other each step operation is same）. testing sample well. add Sample dilution 40μl to testing sample well, then add testing sample 10μl （sample final dilution is 5-fold）, add sample to wells , don’t touch the well wall as far as possible, and Gently mix.
3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.
4.Configurate liquid: 30-fold wash solution diluted 30-fold （or 20-fold） with distilled water and reserve.
5.washing：Uncover Closure plate membrane, discardLiquid, dry by swing, add washing buffer to every well, still for 30s then drain,repeat 5 times, dry by pat.
6.add enzyme：Add HRP-Conjugate reagent 50μl to each well, except blank well.
7.incubate：Operation with 3.
8.washing：Operation with 5.
9.color：Add Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light preservation for 15 min at 37℃
10.Stop the reaction：Add Stop Solution50μl to each well, Stop the reaction（the blue color change to yellow color）.
11.assay：take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min.
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