Matriptase ELISA Assay Kit

$750.00

SKU: MAT31-K01 Categories: , ,
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Matriptase ELISA Assay Kit:

For Research Use Only (RUO)
Size:  1×96 wells
Sensitivity:  2.0 ng/ml
Dynamic Range:  2 – 512 ng/ml
Incubation Time:  2.5 hours
Sample Type:  Serum   
Sample Size: 100 µL

Product Developed and Manufactured in the USA
Intended Use
The Eagle Biosciences Matriptase (TADG-15) ELISA Kit is intended for the quantitative determination of the cancer antigen Matriptase concentration in human serum.  The Matriptase (TADG-15) ELISA assay kit is for research use only and not to be used in diagnostic procedures. 

Assay Principle
The Matriptase (TADG-15) ELISA assay kit is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a monoclonal antibody directed against a distinct antigenic determinant on the intact Matriptase molecule for solid phase immobilization (on the microtiter wells). Standards, calibrators, and patient samples are incubated with the solid phase antibody on the plate. Wells are then washed and incubated with a biotin conjugated anti-Matriptase monoclonal antibody.  After a second wash Streptavidin conjugated to HRP is added as a reporting agent. Excess streptavidin-HRP is then washed off and a solution of TMB Reagent is added and incubated resulting in the development of a blue color if Matriptase is present. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of Matriptase is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

  1. Wash wells one time with 300ul/well 1X PBS, remove liquid from wells, and pat dry on absorbance paper or paper towel.
  2. Dispense 100µl of Matriptase standards, calibrators, and diluted specimens into appropriate wells.
  3. Incubate at room temperature (18-26°C) for 60 minutes with gentle agitation.
  4. Remove samples by emptying the plate contents into a waste container.
  5. Remove liquid from all wells. Wash wells three times with 300µl of 1X wash buffer. Blot on absorbance paper or paper towel after each wash.
  6. Strike the microtiter plate sharply onto absorbance paper or paper towels to remove all residual liquid droplets.
  7. Dispense 100µl of Biotin-labeled Antibody into each well and incubate at room temperature for 60 minutes with gentle agitation.
  8. Repeat steps 5 and 6.
  9. Dispense 100µl Strept-HRP into each well and incubate at room temperature for 30 minutes with gentle agitation.
  10. Repeat steps 5 and 6.
  11. Dispense 100µl of TMB Reagent into each well and incubate at room temperature in the dark for 30 minutes.
  12. Stop the reaction by adding 100µl of Stop Solution into each well.
  13. Gently mix for 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
  14. Read the optical density at 450nm with a microtiter plate reader within 15 minutes.

Assay Background
The protein encoded by the Matriptase gene is an epithelial-derived, integral membrane serine protease. This protease forms a complex with the Kunitz-type serine protease inhibitor, HAI-1, and is found to be activated by sphingosine 1-phosphate. This protease has been shown to cleave and activate hepatocyte growth factor/scattering factor, and urokinase plasminogen activator, which suggest the function of this protease as an epithelial membrane activator for other proteases and latent growth factors. The expression of this protease has been associated with breast, colon, prostate, and ovarian tumors, which implicates its role in cancer invasion and metastasis.