26S Proteasome (26S-PSM) BioAssay™ ELISA Kit (Human)

Kits and Assays / Kits-ELISA (Proteasomes), BioAssay™

4°C/-20°C

26S Proteasome (26S-PSM) BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of 26S-PSM in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.

353541 is specific only for the detection of the 26S Proteasome and was tested with intact 26S proteasome and with cell lysates.  
353541 is not cross-reactive with the following: 19S particle, 20S particle, PSMB1, LMP2 and PSMA3.
The capture antibody is specific for the 20S particle while the detection antibody is specific for the 19S particle.

Detection Range:
0.313-20ng/ml

Sensitivity:
<0.188ng/ml

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

Kit Components:
*353541A: Microtiter Strips, 1x96 wells (8x12 wells).
*353541B: Standard, 2x1 vial
353541C: Sample/Standard Dilution Buffer, 1x20ml
353541D: Antibody (Biotin) (Concentrated), 1x120ul
353541E: Antibody Dilution Buffer, 1x10ml
353541F: Streptavidin (HRP) (SABC), 1x120ul
353541G: SABC Dilution Buffer, 1x10ml
353541H: TMB Substrate, 1x10ml
353541J: Stop Solution, 1x10ml
353541K: Wash Buffer, 25X, 1x30ml

Storage and Stability:
Store unopened *353541A at 4ºC; store at -20ºC once opened. Store unopened *353541B 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 Principle:
This ELISA kit employs the sandwich enzyme-linked immunoassay technique, utilizing a microtiter plate pre-coated with an antibody specific to 26S-PSM. Standards and samples are added to the appropriate wells, then incubated. Aspirate and wash the plate. 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 then washed. Then the TMB substrate solution is added and incubated. After the TMB substrate solution is added, only those wells that contain 26S-PSM, the antibody (Biotin) and Streptavidin (HRP) 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 26S-PSM in the sample is then determined by comparing the O.D. of the sample to the standard curve.

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.