α1AC, AACT, ALPHA 1-ANTICHYMOTRYPSIN ELISA KIT (1GNH 4.556 - 6.925 ug/mL Sensitivity, 5 - 1600 ug/mL Assay Range: Part m, r, c AACT-ELISA)

• Assay ranges 4.556 µg/mL to 6.925 µg/mL depending on species.
• 5 µg/mL - 1600 µg/mL sensitivity depending on species
• Separate kits are available for mouse, rat and canine SERPINA3. 
• Sandwich ELISA format providing convenience and high sensitivity. 
• Strip plate format allowing separate use and storage of sections of the ELISA plate.
• Rapid protocol:  Only one incubation with sample to antibodies.  Total protocol time approximately 1.5 hrs.
• Colormetric detection at 450nm using a standard absorbance plate reader.

Protocol

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1. 8 x 12 well removable section strip plate coated with anti-CRP antibody
2. Anti-CRP polyclonal detection antibody conjugated with biotin
3. Steptavidin-HRP
4. Protein Standard
5. Standard diluent
6. Concentrated wash solution
7. Chromogen solutions A and B
8. Stop solution
9. Plate sealers

Alpha 1-Antichymotrypsin (A1AC), encoded by the SERPINA3 gene, inhibits proteases like trypsin that are typically released by immune cells as part of the inflammatory response to pathoogens.  a1AC expression may also be correlated with Alzeimer's disease pathogenesis since it enhances the formation of amyloid fibrils.    

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Methods Overview:

The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen.  Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate.  The plate is washed after a 60 min incubation period.  Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution.  A stop solution is applied to terminate the reaction, and the solution is measured at 450nm.  

   Sandwich ELISA Methods                                                                                                  

ELISA Assay Steps

1. Mix sample, biotinylated-detection antibody and streptavidin-HRP.
2. Apply sample and antibody-HRP complex to ELISA plate for 60min at 37^oC.  Wash 5 times.
3. Apply chromogens, incubate, then stop reaction.