Beta-1,4-galactosyltransferase 1 (B4galt1) BioAssay™ ELISA Kit (Mouse)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Beta-1, 4-galactosyltransferase 1 (B4galt1) BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of B4galt1 in mouse serum, plasma, tissue homogenates and other biological fluids.

Detection Range:
78-5000pg/ml

Sensitivity:
46.9pg/ml

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

Assay Principle:
This ELISA kit employs the sandwich enzyme-linked immunoassay technique, utilizing a microtiter plate pre-coated with an antibody specific to B4galt1. Standards and samples are added to the appropriate wells, then the Antibody (Biotin) is added to all the wells. The plate is incubated and then washed. Streptavidin (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain B4galt1, 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 B4galt1 in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*354169A: Microtiter Strips, 1x96 wells (8x12 wells).
*354169B: Standard, 2x1 vial
354169C: Sample/Standard Dilution Buffer, 1x20ml
354169D: Antibody (Biotin) (Concentrated), 1x120ul
354169E: Antibody Dilution Buffer, 1x10ml
354169F: Streptavidin (HRP) (SABC), 1x120ul
354169G: SABC Dilution Buffer, 1x10ml
354169H: TMB Substrate, 1x10ml
354169J: Stop Solution, 1x10ml
354169K: Wash Buffer, 25X, 1x30ml

Storage and Stability:
Store unopened *354169A at 4ºC; store at -20ºC once opened. Store unopened *354169B 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 15-30 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results