0.313-20ng/ml

0.188ng/ml

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

The microtiter plate provided in this kit has been pre-coated with an antibody specific to HSPG. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to HSPG. 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 HSPG, 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 HSPG in the sample is then determined by comparing the O.D. of the sample to the standard curve.

*356400A: Microtiter Strips, 1x48 wells or 1x96 wells *356400B: Standard, 1x1vial or 2x1vial 356400C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml 356400D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul 356400E: Antibody Dilution Buffer, 1x5ml or 1x10ml 356400F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul 356400G: SABC Dilution Buffer, 1x5ml or 1x10ml 356400H: TMB Substrate, 1x5ml or 1x10ml 356400J: Stop Solution, 1x5ml or 1x10ml 356400K: Wash Buffer, 25X, 1x15ml or 1x30ml

Store unopened *356400A at 4ºC; store at -20ºC once opened. Store unopened *356400B 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.

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.