Glycosylated Hemoglobin/Hemoglobin A1c (HbA1C ) BioAssay™ ELISA Kit (Mouse)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Glycosylated Hemoglobin/Hemoglobin A1c (HbA1C) BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of HbA1C in mouse serum, plasma, tissue homogenates and other biological fluids.

Detection Range:
1.25-80ng/ml

Sensitivity:
0.75ng/ml

Precision:
Intra-Assay: CV<8%
Inter-Assay: CV<10%

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to HbA1C. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to HbA1C. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain HbA1C, Biotin-conjugated antibody and enzyme-conjugated Streptavidin complex will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of HbA1C in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*356204A: Microtiter Strips, 1x48 wells or 1x96 wells
*356204B: Standard, 1x1vial or 2x1 vial
356204C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml
356204D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul
356204E: Antibody Dilution Buffer, 1x5ml or 1x10ml
356204F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul
356204G: SABC Dilution Buffer, 1x5ml or 1x10ml
356204H: TMB Substrate, 1x5ml or 1x10ml
356204J: Stop Solution, 1x5ml or 1x10ml
356204K: Wash Buffer, 25X, 1x15ml or 1x30ml

Storage and Stability:
Store unopened *356204A at 4ºC; store at -20ºC once opened. Store unopened *356204B at 4°C; once reconstituted, store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.

Assay Summary:
1. Wash plate 2 times before adding standards and samples to wells!
2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C. Aspirate and wash 2 times.
3. Add 100ul Antibody (Biotin) working solution to each well and incubate for 60 minutes at 37°C.
4. Aspirate and wash 3 times.
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C
6. Aspirate and wash 5 times.
7. Add 90ul TMB substrate. Incubate 10-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results