Carboxymethyl Lysine (CML) BioAssay™ ELISA Kit (Human)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Carboxymethyl Lysine (CML) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of CML in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.

Detection Range:
15.6-1000ng/ml

Sensitivity:
<9.4ng/mlg/ml

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

Assay Principle:
This ELISA kit employs the quantitative competitive enzyme-linked immunoassay technique, utilizing a microtiter plate provided in this kit has been pre-coated with CML. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to CML. 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 CML, Biotin-conjugated antibody and enzyme-conjugated Streptavidin 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 CML in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*354848A: Microtiter Strips, 1x48 wells or 1x96 wells
*354848B: Standard, 1x1vial or 2x1 vials
354848C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml
354848D: Antibody (Biotin) (Concentrated), 1x30ul or 1x60ul
354848E: Antibody Dilution Buffer, 1x5ml or 1x10ml
354848F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul
354848G: SABC Dilution Buffer, 1x5ml or 1x10ml
354848H: TMB Substrate, 1x5ml or 1x10ml
354848J: Stop Solution, 1x5ml or 1x10ml
354848K: Wash Buffer, 25X, 1x15ml or 1x30ml

Storage and Stability:
Store unopened *354848A at 4ºC; store at -20ºC once opened. Store unopened *354848B 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 50ul standard or sample to each well.
3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 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 15-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results.