詳細介紹
人LF/LTF Ab-Ig ELISA試劑盒
產(chǎn)品別名:人乳鐵傳遞蛋白/乳鐵蛋白抗體IgG ELISA試劑盒
英文名稱:Human Lactoferrin antibody IgG,LF/LTF Ab-IgG ELISA KIT
產(chǎn)品規(guī)格:96T/48T
操作步驟
1.標準品的稀釋:本試劑盒提供原倍標準品一支,用戶可按照下列圖表在小試管中進行稀釋。
80μg/L5號標準品150μl的原倍標準品加入150μl標準品稀釋液
40μg/L4號標準品150μl的5號標準品加入150μl標準品稀釋液
20μg/L3號標準品150μl的4號標準品加入150μl標準品稀釋液
10μg/L2號標準品150μl的3號標準品加入150μl標準品稀釋液
5μg/L1號標準品150μl的2號標準品加入150μl標準品稀釋液
2.加樣:分別設空白孔(空白對照孔不加樣品及酶標試劑,其余各步操作相同)、標準孔、待測樣品孔。在酶標包被板上標準品準確加樣50μl,待測樣品孔中先加樣品稀釋液40μl,然后再加待測樣品10μl(樣品zui終稀釋度為5倍)。加樣將樣品加于酶標板孔底部,盡量不觸及孔壁,輕輕晃動混勻。
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5次,拍干。
6.加酶:每孔加入酶標試劑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(此時藍色立轉(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)。 測定應在加終止液后15分鐘以內(nèi)進行。
Assay procedure
1.Dilute and add sample:Dilute Original density Standard as follow table:
800ng/ml5 Standard150μl Original density Standard+150μl Standard diluent
400ng/ml4 Standard150μl 5 Standard+150μl Standard diluent
200ng/ml3 Standard150μl 4 Standard+150μl Standard diluent
100ng/ml2 Standard150μl 3 Standard +150μl Standard diluent
50ng/ml1 Standard150μl 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, discard Liquid, 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.
LF,人乳鐵傳遞蛋白,乳鐵蛋白抗體IgG