MMP-7 ELISA Assay Kit

$750.00

The Eagle Biosciences MMP-7 ELISA Kit is intended for the quantitative determination of the Cancer Antigen MMP-7 concentration in human serum. The Matrix Metalloproteinase 7 (MMP-7) ELISA assay kit is for research use only and not to be used in diagnostic procedures.

SKU: MM731-K01 Categories: , ,

MMP-7 ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 1.0 ng/mL
Dynamic Range: 4 – 64 ng/mL
Incubation Time: 2.25 hours
Sample Type: Serum
Sample Size: 50 µL

Product Developed and Manufactured in the USA


Additional Information

Assay Background

MMP-7 proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP’s are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The MMP-7 enzyme degrades proteoglycans, fibronectin, elastin and casein and differs from most MMP family members in that it lacks a conserved C-terminal protein domain. The enzyme is involved in wound healing, and studies in mice suggest that it regulates the activity of defensins in intestinal mucosa. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.

Assay Principle

The Eagle Biosciences MMP-7 ELISA test 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 MMP-7 molecule for solid phase immobilization (on the microtiter wells). Standards, calibrators, and samples are incubated with the biotin conjugated anti-MMP-7 monoclonal antibody and the solid phase antibody on the plate simultaneously. Wells are then washed and incubated with Streptavidin conjugated to HRP which is used 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 MMP-7 is present. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of MMP-7 is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

Assay Procedure

Bring all specimens and MMP-7 ELISA assay kit reagents to room temperature (18-26°C) and gently mix. If precipitate is seen in standards, vortex vigorously after reaching room temperature to re-suspend standard.

  1. Thaw serum samples and bring to room temperature. Briefly vortex or mix samples.
  2. Dilute serum samples 1:4 in 1X PBS before being added to the plate.
  3. Dispense 50µl of MMP-7 standards, calibrators, and specimens into appropriate wells.
  4. Dispense 50µl of Biotin-labeled Antibody into each well and incubate at 30°C for 1 hour at lowest rpm in shaking incubator.
  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 Streptavidin-HRP into each well and incubate at room temperature for 30 minutes with gentle agitation.
  8. Repeat steps 5 and 6.
  9. Dispense 100µl TMB Substrate into each well and incubate at room temperature in the dark for 30 minutes.
  10. Stop the reaction by adding 100µl of Stop Solution into each well.
  11. Gently mix for 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
  12. Read the optical density at 450nm with a microtiter plate reader within 15 minutes.

Typical Standard Curve

Manual

Product Manual