Hepc (Hepcidin) BioAssay™ ELISA Kit (Rat)

Kits and Assays / Kits-ELISA (Antimicrobial Peptides), BioAssay™

4°C/-20°C

Hepc (Hepcidin) BioAssay™ ELISA Kit is for the quantitative detection of Hepc in rat serum, plasma, tissue homogenates and other biological fluids.

Range:
78.125-5000pg/ml

Sensitivity:
<46.875pg/ml

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

Assay Principle:
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Hepc antibody was pre-coated onto 96-well plates. And the biotin conjugated Hepc antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and washed with wash buffer. HRP-Streptavidin was added and unbound conjugates were washed away with wash buffer. TMB substrate was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Hepc amount of sample captured on plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Hepc can be calculated.

Kit Components:
*35654A: Microtiter Strips, 1x48 wells or 1x96 wells
*35654B: Standard, 1x1vial or 2x1vial
35654C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml
35654D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul
35654E: Antibody Dilution Buffer, 1x5ml or 1x10ml
35654F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul
35654G: SABC Dilution Buffer, 1x5ml or 1x10ml
35654H: TMB Substrate, 1x5ml or 1x10ml
35654J: Stop Solution, 1x5ml or 1x10ml
35654K: Wash Buffer, 25X, 1x15ml or 1x30ml

Storage and Stability:
Store unopened *35654A at 4ºC. Once opened, store at -20ºC. Store *35654B powder 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. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Assay Summary:
1. Wash plate 2 times before adding standards, samples and control (zero) to wells!
2. Add 100ul standard or sample to each well for 90 minutes at 37°C
3. add 100ul Biotin antibody working solution to each well 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. Calculation of results