MMP12 ELISA Kit

Catalog No. ABIN1672891

Product Details MMP12 ELISA Kit

Target: MMP12 (Matrix Metallopeptidase 12 (Macrophage Elastase) (MMP12))

Binding Specificity: AA 18-462

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Minimum Detection Limit: 62.5 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse MMP-12

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin)

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: A18-C462

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: A18-C462

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the peptidase M10A family.

Plate: Pre-coated

Protocol: mouse MMP-12 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for MMP-12 has been precoated onto 96-well plates. Standards(NSO, A18-C462) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MMP-12 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse MMP-12 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 4000pg/mL, 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL mouse MMP-12 standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse MMP-12 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 337, Standard deviation: 18.2, CV(%): 5.4
• Sample 2: n=16, Mean(pg/ml): 1472, Standard deviation: 92.74, CV(%): 6.3
• Sample 3: n=16, Mean(pg/ml): 2522, Standard deviation: 121.1, CV(%): 4.8,
• Sample 1: n=24, Mean(pg/ml): 426, Standard deviation: 27.69, CV(%): 6.5
• Sample 2: n=24, Mean(pg/ml): 1538, Standard deviation: 109.2, CV(%): 7.1
• Sample 3: n=24, Mean(pg/ml): 2768, Standard deviation: 157.8, CV(%): 5.7

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for MMP12 ELISA Kit (ABIN1672891)

Papakonstantinou, Karakiulakis, Batzios, Savic, Roth, Tamm, Stolz: "Acute exacerbations of COPD are associated with significant activation of matrix metalloproteinase 9 irrespectively of airway obstruction, emphysema and infection." in: Respiratory research, Vol. 16, pp. 78, (2016) (PubMed).

Zang, Zhuang, Deng, Yang, Ye, Xie, Ren, Fu, Luo, Xu, Liu: "Pulmonary C Fibers Modulate MMP-12 Production via PAR2 and Are Involved in the Long-Term Airway Inflammation and Airway Hyperresponsiveness Induced by Respiratory Syncytial Virus Infection." in: Journal of virology, Vol. 90, Issue 5, pp. 2536-43, (2016) (PubMed).

Vaz, Rajasekaran, Potteti, Reddy: "Myeloid-specific Fos-related antigen-1 regulates cigarette smoke-induced lung inflammation, not emphysema, in mice." in: American journal of respiratory cell and molecular biology, Vol. 53, Issue 1, pp. 125-34, (2015) (PubMed).

Target Details for MMP12

Target: MMP12 (Matrix Metallopeptidase 12 (Macrophage Elastase) (MMP12))

Alternative Name: MMP12 (MMP12 Products)

Background:

Protein Function: May be involved in tissue injury and remodeling. Has significant elastolytic activity. Can accept large and small amino acids at the P1' site, but has a preference for leucine. Aromatic or hydrophobic residues are preferred at the P1 site, with small hydrophobic residues (preferably alanine) occupying P3 (By similarity). .

Background: Matrix metalloproteinase-12(MMP12), also known as MME or ME, is an enzyme that in humans is encoded by the MMP12 gene. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.2. It is thought that the protein encoded by this gene is cleaved at both ends to yield the active enzyme, but this processing has not been fully described. The enzyme degrades soluble and insoluble elastin. It may play a role in aneurysm formation and studies in mice suggest a role in the development of emphysema. This gene may involved in tissue injury and remodeling.

Synonyms: Macrophage metalloelastase,MME,3.4.24.65,Matrix metalloproteinase-12,MMP-12,Mmp12,Mme, Mmel,

Full Gene Name: Macrophage metalloelastase

Cellular Localisation: Secreted, extracellular space, extracellular matrix.

Gene ID: 17381

UniProt: P34960