Nori Human MMP-25 ELISA Kit

$461.00$832.00

DataSheet   

This ELISA kit is for quantification of MMP-25 in human. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for MMP-25 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any MMP-25 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for MMP-25 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of MMP-25 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for MMP-25: Matrix metalloproteinase-25 (MMP-25), membrane type 6-MMP (MT6-MMP), MMP-20, MT-MMP-6, leukolysin, MMPL1

This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.

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Description

Nori Human MMP-25 ELISA Kit Summary

Alternative names for MMP-25: Matrix metalloproteinase-25 (MMP-25), membrane type 6-MMP (MT6-MMP), MMP-20, MT-MMP-6, leukolysin, MMPL1

 

Assay Type Solid Phase Sandwich ELISA
Format 96-well Microplate or 96-Well Strip Microplate
Method of Detection Colorimetric
Number of Targets Detected 1
Target Antigen Accession Number Q9NPA2
Assay Length 3 hours
Quantitative/Semiquantitative Quantitative
Sample Type Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL,
Recommended Sample Dilution (Plasma/Serum) No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ
Sensitivity 300 pg/mL
Detection Range 1.563-100 ng/mL
Specificity Human MMP-25
Cross-Reactivity < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested.
Interference No significant interference observed with available related molecules
Storage/Stability 4 ºC for up to 6 months
Usage For Laboratory Research Use Only. Not for diagnostic or therapeutic use.
Additional Notes The kit allows for use in multiple experiments.

 

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Matrix metalloproteinase-25 (MMP-25) is an enzyme that is encoded by the MMP25 gene.[1][2] MMP-25 is also known as membrane type 6-MMP (MT6-MMP), MMP-20, MT-MMP-6, leukolysin, MMPL1. Membrane-type 6 matrix metalloproteinase (MT6-MMP, MMP-25) is the second glycosyl-phosphatidyl inositol (GPI)-anchored MMP.[3] MT6-MMP was highly regulated by cytokine and chemokines in neutrophils.[4]

Proteins of 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 MMPs are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. However, MMP-25 is a member of the membrane-type MMP (MT-MMP) subfamily, attached to the plasma membrane via a glycosylphosphatidyl inositol anchor. In response to bacterial infection or inflammation, MMP-25 is thought to inactivate alpha-1 proteinase inhibitor, a major tissue protectant against proteolytic enzymes released by activated neutrophils, facilitating the trans-endothelial migration of neutrophils to inflammatory sites. MMP-25 is highly expressed in human colon cancer and promotes tumor growth [5] and it may also play a role in tumor invasion and metastasis through activation of MMP2.[6]

References

  1. Pei D (2000). Cell Res. 9 (4): 291–303.
  2. Velasco G, et al. (2000). Cancer Res. 60 (4): 877–82.
  3. Kojima S, Itoh Y, Matsumoto S, et al. (2000) FEBS Lett. 480 (2–3): 142–6.
  4. Kang T, Yi J, Guo A, et al. (2001). J. Biol. Chem. 276 (24): 21960–8.
  5. Sun Q, Weber CR, Sohail A, et al. (2007). J. Biol. Chem. 282 (30): 21998–2010.
  6. Nie J, Pei D (2003). Cancer Res. 63 (20): 6758–62.

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