BD-EL-H1527 人巨噬细胞炎性蛋白2(MIP-2)酶联免疫吸附测定试剂盒

Standard microplate reader ( filter with detection wavelength of 450nm , filter with calibration wavelength of 570nm or 630nm ).

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1.Determine the number of plates needed according to the number of samples to be tested and the number of standard product. Each standard product and blank hole is recommended to make multiple holes. The number of samples can be decided by users, and make multiple holes as far as possible.

2. Sample adding: 1) :( blank holes: sample, biotin-labeled anti -GLP-7-37 antibodies, streptavidin-HRP should not be added into the blank holes, and other stepsare the same; 2) standard product hole: Add standard product 50μl , streptavidin--HRP50μl (standard products have integrated well in advance the biotinylated antibody, it is not added at the time of operation, and only add streptomycin -HRP); 3) sample action holes: Add sample 40μl, then each added anti - GLP-7-37 antibody 10μl, and 50μl of streptavidin-HRP (except blank holes). Cover with the seal plate membrane, and mix gently by shaking, incubate in 37 ℃ temperature for 60 minutes.

3. Dosing liquid: dilute 30-time concentrated washer with distilled water in 30 times for use, and 20-time concentrated washer with distilled water in 20 times for use.

4. Washing: Uncover the plate membrane, remove liquid, then dry it up, add washing liquid to fill each hole, after stewing for 30 seconds then remove it, repeat for 5 times, pat dry.

5. coloration: add each hole with the chromogenic reagent A 50 μl, then add reagent B 50μl, gently shake to mix, keep in 37 ℃ in dark for 10 minutes.

6. Stop solution: each hole add stop solution 50μl, then stop the reaction (the blue color change into yellow immediately).

7. Determination: adjust blank hole to zero, wavelength of 450nm sequentially measured the absorbance of each well (OD value). Determination should be conducted within 10 minutes after adding the stop solution.

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