Kallikrein Related Peptidase 8 (KLK8) ELISA

$750.00

SKU: KL831-K01 Categories: , ,
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Kallikrein-Related Peptidase 8 (KLK8) ELISA:
For Research Use Only (RUO)
Size:  1×96 wells
Sensitivity:  1.0 ng/ml
Dynamic Range:  1 – 256 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 Kallikrein-Related Peptidase 8 (KLK8) ELISA Kit is intended for the quantitative determination of the Kallikrein-Related Peptidase 8 (KLK8) concentration in human serum.  The Kallikrein-Related Peptidase 8 (KLK8) ELISA assay kit is for research use only and not to be used in diagnostic procedures.

Assay Principle
The KLK8 (TADG-14) ELISA assay kit is based on the principle of a solid phase enzyme-linked immunosorbent assay (ELISA). The assay system utilizes a monoclonal antibody directed against a distinct antigenic determinant on the intact KLK8 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 an anti-KLK8polyclonal antibody.  After a second wash Goat anti-Rabbit antibody conjugated to HRP is added as a reporting agent. Excess Goat anti-Rabbit -HRP is then washed off and a solution of TMB Reagent is added and incubated resulting in the development of a blue color if KLK8 is present. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of KLK8 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 KLK8 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 Polyclonal Antibody into each well and incubate at room temperature for 60 minutes with gentle agitation.
  8. Repeat steps 5 and 6.
  9. Dispense 100µl Goat anti-Rabbit-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
Kallikreins are a subgroup of serine proteases having diverse physiological functions. Growing evidence suggests that many kallikreins are implicated in carcinogenesis and some have potential as novel cancer and other disease biomarkers. KLK8 is one of the fifteen kallikrein subfamily member. The KLK8 enzyme exhibits distinct patterns of expression that suggest roles in brain plasticity and ovarian cancer.