MMP-9 Immunoassay Kit, proform only

MMP-9 Immunoassay Kit, proform only

Chemicon (Millipore)

100 assays

The MMP-9 immunoassay kit utilizes antibodies immobilized on a bead matrix, in combination with enzyme-labeled antibodies, directed against different antigenic sites on the same MMP-9 molecule. Upon the addition of an MMP-9-containing specimen, the MMP-9 molecule is sandwiched between the antibody coated on the bead and enzyme-labeled antibodies. After removing unbound enzyme-labeled antibody, the bead containing the MMP-9 complex is incubated with enzyme substrate and o-phenylenediamine, resulting in the development of color. The resultant color enzyme is proportional to the amount of MMP-9 present, the concentration of MMP-9 in specimens can be determined from a standard curve.

Test Principle:

The MMP-9 immunoassay kit utilizes antibodies immobilized on a bead matrix, in combination with enzyme-labeled antibodies, directed against different antigenic sites on the same MMP-9 molecule. Upon the addition of an MMP-9-containing specimen, the MMP-9 molecule is sandwiched between the antibody coated on the bead and enzyme-labeled antibodies. After removing unbound enzyme-labeled antibody, the bead containing the MMP-9 complex is incubated with enzyme substrate and o-phenylenediamine, resulting in the development of color. The resultant color enzyme is proportional to the amount of MMP-9 present, the concentration of MMP-9 in specimens can be determined from a standard curve.

Application:

The MMP-9 immunoassay kit is useful for the determination of MMP-9 (pro-MMP-9) levels in fresh human plasma and conditioned medium of human cells. Human serum is not an acceptable sample for evaluation.

This system recognizes free pro-MMP-9, intermediate 83 kDa MMP-9, and MMP-9 in complex with TIMP-1 with the same efficiency. The assay does not recognize active MMP-9 (67 kDa). Contents of this kit are sufficient for assay of 100 samples, including standard curve. Testing of samples in duplicate or triplicate is strongly recommended. This kit is intended for research use only; not for diagnostic or therapeutic applications.

Activity Assay

Human Only

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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. However, the enzyme encoded by this gene is stored in secondary granules within neutrophils and is activated by autolytic cleavage. Its function is degradation of type I, II and III collagens. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.

FUNCTION: SwissProt: P22894 # Can degrade fibrillar type I, II, and III collagens.
COFACTOR: Binds 3 calcium ions per subunit. & Binds 2 zinc ions per subunit.
SIZE: 467 amino acids; 53412 Da
SUBCELLULAR LOCATION: Cytoplasmic granule. Note=Stored in intracellular granules.
TISSUE SPECIFICITY: Neutrophils.
DOMAIN: SwissProt: P22894 The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
SIMILARITY: Belongs to the peptidase M10A family. & Contains 4 hemopexin-like domains.

• Anti-MMP-9 Coated Beads - One (1) bottle containing 100 anti-MMP-9 coated polystyrene beads.
• Concentrated Enzyme Labeled Antibody Solution - One (1) 1.2 mL bottle containing MMP-9 antibody.
• Color Reagent (Lyophilized) - Four (4) bottles o-Phenylenediamine (OPD), each sufficient to make one 12 mL bottle of color reagent.
• Substrate Solution - One (1) 50 mL bottle 0.02% Hydrogen Peroxide.
• Stop Solution - Two (2) bottles, sufficient to make 100 mL 1.75 N Sulfuric acid per bottle.
• Buffer Reagent (Lyophilized) - One (1) 50 mL bottle BSA in sodium phosphate buffer.
• MMP-9 Standard (Lyophilized) - Two (2) bottles, sufficient to make 0.5 mL Human pro-MMP-9 per bottle.
• Concentrated Washing Solution - Two bottles, sufficient to make 1L Sodium phosphate buffer, pH 7.0, per bottle.

Chemicon

This product is not available for sale in Japan.

P22894

Store kit at 2-8°C.

Precautions:

· All specimens should be handled as potentially infectious.

· Do not freeze reagents

· Do not combine reagents from different lots.

· Stop solution (1.75 N Sulfuric acid) should be handled with caution.

· Do not allow Color or Stop Solution to come into contact with metal.

· Do not use plasma which were frequently frozen and thawed.

· All reagents should be brought to ambient temperature before use. Gently stir each bottle of liquid reagent. Do not shake reagent bottles.

· Use pipette with disposable tips for pipetting standards and specimens.

• MMP8
• HNC
• CLG1
• PMNL-CL
• MMP-8
• EC 3.4.24.34

MMP-9 Immunoassay Kit, proform only

NM_002424.1

· Pipette (10 mL)

· Pipettors & tips capable of accurately measuring 50-1000 μL

· Dispensor (3 mL)

· Test tubes (internal diameter 10-13 mm; length 60-75 mm)

· Aspirator

· Graduated cylinder (50 mL, 1000 mL)

· Spectrophotometer (492 nm)