LEUKOCYTE ELASTASE (ELA2, LE) ELISA KIT (3Q76, 0.08 - 0.185 ng/mL Sensitivity, 0.08 - 42 ng/mL Assay Range: Part LE-ELISA, m r or c)

• Assay ranges 0.08 ng/mL to 42 ng/mL depending on species.
• 0.05 ng/mL - 0.185 ng/mL sensitivity depending on species
• Separate kits are available for mouse, rat and canine CRP. 
• 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-ELA2 antibody
2. Anti-LE 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

Leukocyte Elastase (ELANE, ELA2, HLE) aka Neutrophil Elastase is a very non-specific serine proteinase secreted by neutrophils to hydrolyze the cell surface proteins of bacteria.  However, ELANE can also hydrolyze a variety of other protein substrates, including structural collagen and elastin due to its catalytic triad, contributing to degenerative, inflammatory diseases like emphysema in the lung and perhaps Alzheimer's Disease, when it targets otherwise healthy tissue.  Lack of functional ELA2 can cause congenital neutropenia since the neutrophils cannot mature.

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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.