Recombinant Anti-MMP2 antibody [EPR1184] (ab92536)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1184] to MMP2
- Suitable for: Flow Cyt (Intra), ICC/IF, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-MMP2 antibody [EPR1184]
See all MMP2 primary antibodies -
Description
Rabbit monoclonal [EPR1184] to MMP2 -
Host species
Rabbit -
Specificity
In Western Blot, this product typically gives a weaker signal in some hepatocarcinoma like HepG2, MHCC97L(PMID:33184263 ) et al. Please use reconmended positive control when testing these cells.
Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human MMP2 aa 550-650 (C terminal). The exact sequence is proprietary.
Database link: P08253 -
Positive control
- WB: HepG2, L6, Raw264.7 and NIH/3T3 cell lysates; fetal heart and human skin tissue lysate; Human plasma, brain and breast tissue lysate ICC/IF: PC-3 cells Flow Cyt (intra): HeLa and PC-3 cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1184 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92536 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/400.
For unpurified, use 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
1/250.
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WB | (4) |
1/1000 - 1/5000. Predicted molecular weight: 74 kDa.
For Lysate preparation protocol, please refer to the protocol here (downloadable copy). Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot. |
Notes |
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Flow Cyt (Intra)
1/400. For unpurified, use 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/250. |
WB
1/1000 - 1/5000. Predicted molecular weight: 74 kDa. For Lysate preparation protocol, please refer to the protocol here (downloadable copy). Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot. |
Target
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Function
Ubiquitinous metalloproteinase that is involved in diverse functions such as remodeling of the vasculature, angiogenesis, tissue repair, tumor invasion, inflammation, and atherosclerotic plaque rupture. As well as degrading extracellular matrix proteins, can also act on several nonmatrix proteins such as big endothelial 1 and beta-type CGRP promoting vasoconstriction. Also cleaves KISS at a Gly-
-Leu bond. Appears to have a role in myocardial cell death pathways. Contributes to myocardial oxidative stress by regulating the activity of GSK3beta. Cleaves GSK3beta in vitro.
PEX, the C-terminal non-catalytic fragment of MMP2, posseses anti-angiogenic and anti-tumor properties and inhibits cell migration and cell adhesion to FGF2 and vitronectin. Ligand for integrinv/beta3 on the surface of blood vessels. -
Tissue specificity
Produced by normal skin fibroblasts. PEX is expressed in a number of tumors including gliomas, breast and prostate. -
Involvement in disease
Defects in MMP2 are the cause of Torg-Winchester syndrome (TWS) [MIM:259600]; also known as multicentric osteolysis nodulosis and arthropathy (MONA). TWS is an autosomal recessive osteolysis syndrome. It is severe with generalized osteolysis and osteopenia. Subcutaneous nodules are usually absent. Torg-Winchester syndrome has been associated with a number of additional features including coarse face, corneal opacities, patches of thickened, hyperpigmented skin, hypertrichosis and gum hypertrophy. However, these features are not always present and have occasionally been observed in other osteolysis syndromes. -
Sequence similarities
Belongs to the peptidase M10A family.
Contains 3 fibronectin type-II domains.
Contains 4 hemopexin-like domains. -
Domain
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. -
Post-translational
modificationsPhosphorylation on multiple sites modulates enzymatic activity. Phosphorylated by PKC in vitro.
The propeptide is processed by MMP14 (MT-MMP1) and MMP16 (MT-MMP3). Autocatalytic cleavage in the C-terminal produces the anti-angiogenic peptide, PEX. This processing appears to be facilitated by binding integrinv/beta3. -
Cellular localization
Secreted > extracellular space > extracellular matrix. Membrane. Nucleus. Colocalizes with integrin alphaV/beta3 at the membrane surface in angiogenic blood vessels and melanomas. Found in mitochondria, along microfibrils, and in nuclei of cardiomyocytes. - Information by UniProt
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Database links
- Entrez Gene: 4313 Human
- Entrez Gene: 17390 Mouse
- Entrez Gene: 81686 Rat
- Omim: 120360 Human
- SwissProt: P08253 Human
- SwissProt: P33434 Mouse
- SwissProt: P33436 Rat
- Unigene: 513617 Human
see all -
Alternative names
- 72 kDa gelatinase antibody
- 72kD type IV collagenase antibody
- CLG 4 antibody
see all
Images
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All lanes : Anti-MMP2 antibody [EPR1184] (ab92536) at 1/1000 dilution
Lane 1 : Human plasma tissue lysate
Lane 2 : Human brain tissue lysate
Lane 3 : Human breast tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 74 kDa
Observed band size: 69,72 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot.
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All lanes : Anti-MMP2 antibody [EPR1184] (ab92536) at 1/1000 dilution
Lane 1 : HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate
Lane 2 : Untreated HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 300ng/ml BFA for 24 hours whole cell lysate
Lane 4 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 5 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 74 kDa
Observed band size: 69,72 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking and diluting buffer and concentration: 5% NFDM /TBST.
ab181602 was used as GAPDH loading control.
Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot.
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All lanes : Anti-MMP2 antibody [EPR1184] (ab92536) at 1/1000 dilution
Lane 1 : L6 (Rat skeletal muscle myoblast) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
Lane 2 : Mouse liver lysates prepared in RIPA lysis method with 5% NFDM/TBST
Lane 3 : Mouse liver lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
Lane 4 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST
Lane 5 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
Lane 6 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method with 5% NFDM/TBST
Lane 7 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 74 kDa
Observed band size: 69,72 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThe 72 KDa band is pro-MMP2 and the 69 KDa band is active-MMP2 reported by PMID 11489818 and 22190701.
This antibody shows low affinity in detecting mouse liver and HepG2 lysates which are positive for MMP2 reported by PMID 24096707 and 24297510.
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Immunofluorescence staining of PC-3 cells with purified ab92536 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92536 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Intracellular Flow Cytometry analysis of PC-3 (human prostate adenocarcinoma) cells labeling MMP2 with purified ab92536 at 1/180 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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All lanes : Anti-MMP2 antibody [EPR1184] (ab92536) at 1/10000 dilution (purified)
Lane 1 : L6 cell lysate
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 74 kDa
Observed band size: 64,72 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST72kDa: propeptide; 64kDa: active form
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All lanes : Anti-MMP2 antibody [EPR1184] (ab92536) at 1/1000 dilution (unpurified)
Lane 1 : L6 cell lysate
Lane 2 : Fetal heart lysate
Lane 3 : NIH/3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 74 kDa -
Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab92536 at a dilution of 1 in 400 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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Anti-MMP2 antibody [EPR1184] (ab92536) at 1/5000 dilution (purified) + human skin at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 74 kDa
Observed band size: 64,72 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST72kDa: propeptide; 64kDa: active form
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (305)
ab92536 has been referenced in 305 publications.
- Zheng Y et al. LncNAP1L6 activates MMP pathway by stabilizing the m6A-modified NAP1L2 to promote malignant progression in prostate cancer. Cancer Gene Ther 30:209-218 (2023). PubMed: 36195720
- Li X et al. Low temperature plasma suppresses proliferation, invasion, migration and survival of SK-BR-3 breast cancer cells. Mol Biol Rep 50:2025-2031 (2023). PubMed: 36538172
- Wang M et al. Isorhamnetin inhibits progression of ovarian cancer by targeting ESR1. Ann Transl Med 10:1216 (2022). PubMed: 36544694
- Tu Y et al. Molecular Imaging of Matrix Metalloproteinase-2 in Atherosclerosis Using a Smart Multifunctional PET/MRI Nanoparticle. Int J Nanomedicine 17:6773-6789 (2022). PubMed: 36600879
- Wu Q et al. UBR5 promotes migration and invasion of glioma cells by regulating the ECRG4/NF-ҡB pathway. J Biosci 47:N/A (2022). WB ; Human . PubMed: 36226349