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Product Name MMP12 Chinese Name 基质金属蛋白酶-12Recombinant rabbit monoclonal anti Alias matrix metalloproteinase 12; MMP12; EC 3.4.24.65; HME; Macrophage elastase; Macrophage metaloelastase; Matrix metalloprotease 12; ME; MGC138506; MME; MMP 12; MMP12_HUMAN; MMP 12; MMP-12; MMP12. Immunogen Species Rabbit Clonality Monoclonal Clone NO. 1A4 React Species Human, Mouse, (predicted: Rat, ) Applications WB=1:1000-2000 IHC-P=1:50-200 IHC-F=1:50-200 Flow-Cyt=1:50 ICC=1:50-200 IF=1:50-200 (Paraffin sections need antigen repair)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.Theoretical molecular weight 42kDa Cellular localization Extracellular matrix Secretory protein Form Liquid Concentration 1mg/ml immunogen KLH conjugated synthetic peptide derived from human MMP12 Lsotype IgG Purification affinity purified by Protein A Buffer Solution 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Storage Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. Attention This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. PubMed PubMed Product Detail 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, metastasis, and atherosclerosis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases.
MMP12 was first described in murine macrophages, later in human macrophages, and more recently in other cell types. Also known as metalloelastase, MMP12 is able to degrade elastin, entactin, laminin 1, fibronectin, type IV collagen as well as insulin B-chain and casein. MMP12 is often confused with the Serine proteinase, Leukocyte elastase (EC 3.4.21.37) because of similar nomenclature. MMP12 is structurally similar to the classical MMPs (MMP1, MMP3); it contains a propeptide with autoinhibitory cysteine switch site, a well-conserved zinc site, hinge region and hemopexin domain. MMP12 lacks a transmembrane domain and furin cleavage site. The zymogen for MMP-12 is about 54 kD, and is quickly activated to the 45 kD form; and this breaks down to cascade of active forms, ending with the common 22 kD form. Stimulated macrophages produce MMP12; it has also been found in osteosarcoma cells, synovial fibroblasts and lung fibroblasts.
Subcellular Location:
Secreted, extracellular space, extracellular matrix (Probable).
Tissue Specificity:
Found in alveolar macrophages but not in peripheral blood monocytes.
Similarity:
Belongs to the peptidase M10A family.
Contains 4 hemopexin-like domains.
SWISS:
P39900
Gene ID:
4321
Database links:Entrez Gene: 4321 Human
Entrez Gene: 17381 Mouse
Omim: 601046 Human
SwissProt: P39900 Human
SwissProt: P34960 Mouse
Unigene: 1695 Human
Unigene: 2055 Mouse
Unigene: 33193 Rat
Product Picture Sample:
Lane 1: A549 cell lysate
Lane 2: Hela cell lysate
Lane 3: MCF-7 cell lysate
Lane 4: THP-1 cell lysate
Primary: Anti-MMP12 (SLM-52292R) at 1:500 dilution
Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution
Predicted band size: 42 kD
Observed band size: 38 kD
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: 1: A431, 2: U87-MG
Protein loading quantity: 20 μg
Exposure time: 10 s
Predicted MW: 88 kDa
Observed MW: 120, 130 kDa
Paraformaldehyde-fixed, paraffin embedded (mouse lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (MMP12) Monoclonal Antibody, Unconjugated (SLM-52292R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (MMP12) Monoclonal Antibody, Unconjugated (SLM-52292R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control:A549.
Primary Antibody (green line): Rabbit Anti-MMP12 antibody (SLM-52292R)
Dilution: 1:50;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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