a1-PI, ALPHA-1-PROTEINASE 1 INHIBITOR ELISA KIT (1KCT 6.635mg/L Sensitivity, 10 - 1500 mg/L Assay Range: Part a-1-PI-ELISA, m r or c)

• Assay ranges 10 mg/L to 1500 mg/L depending on species.
• 6.635 mg/L sensitivity
• Separate kits are available for mouse, rat and canine A1-PI. 
• 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-alpha-1-P inhibitor antibody
2. Anti-A1-PI 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 Proteinase Inhibitor (A1-PI), also known as Alpha-1 Antitrypsin (A1AT) is a protective protease inhibitor that prevents tissues from  being broken down by proteases involved in acute inflammation, including trypsin. It is encoded by the SERPINA gene. Deficiencies in this gene are implicated in hereditary chronic tissue breakdown diseases, particularly in the lung where it causes pulmonary emphysema, and in the synovial fluid of joints where the presence of antibodies against A1PI is associated with rheumatoid arthritis. Alpha-1 Antitrypsin is also a possible indicator in inflammation for Alzheimer's disease.   

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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.