Anti-MMP2 antibody [4D3] (ab2462)

Reviews (3)Q&A (11)References (14)

Overview

  • Product name
    Anti-MMP2 antibody [4D3]
    See all MMP2 primary antibodies
  • Description
    Mouse monoclonal [4D3] to MMP2
  • Host species
    Mouse
  • Specificity
    Binds to human MMP-2 and pro-MMP2. The antibody was tested for cross-reactivity with MMP-1, MMP-3, and MMP-9 and showed mild cross-reactivity with MMP-3.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Rabbit, Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human MMP2 aa 557-569 (internal sequence).
    Sequence:

    TSLGLPPDVQRVD


    Database link: P08253

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • Colon carcinoma. Placental tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab2462 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
ICC/IF Use at an assay dependent concentration. PubMed: 19910495
WB Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Target

  • 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
    modifications
    Phosphorylation 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
  • Database links
    see all
  • Alternative names
    • 72 kDa gelatinase antibody
    • 72kD type IV collagenase antibody
    • CLG 4 antibody
    • CLG 4A antibody
    • CLG4 antibody
    • CLG4A antibody
    • Collagenase Type 4 alpha antibody
    • Collagenase type IV A antibody
    • Gelatinase A antibody
    • Gelatinase alpha antibody
    • Gelatinase neutrophil antibody
    • Matrix metallopeptidase 2 gelatinase A 72kDa gelatinase 72kDa type IV collagenase antibody
    • Matrix metalloproteinase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase) antibody
    • Matrix Metalloproteinase 2 antibody
    • Matrix metalloproteinase II antibody
    • Matrix metalloproteinase-2 antibody
    • MMP 2 antibody
    • MMP II antibody
    • MMP-2 antibody
    • MMP2 antibody
    • MMP2_HUMAN antibody
    • MONA antibody
    • Neutrophil gelatinase antibody
    • PEX antibody
    • TBE 1 antibody
    • TBE-1 antibody
    see all

Images

  • Western blot - Anti-MMP2 antibody [4D3] (ab2462)
    Western blot - Anti-MMP2 antibody [4D3] (ab2462)

    400 ng samples of recombinant human pro-enzyme (lane 1 = MMP-1; lane 2 = MMP-3; lane 3 = MMP-2; lane 4 = MMP-9) were subjected to SDS-PAGE, transferred to PVDF membrane, and detected by immunoblotting with ab2462 specific for MMP-2.

  • Western blot - Anti-MMP2 antibody [4D3] (ab2462)
    Western blot - Anti-MMP2 antibody [4D3] (ab2462)This image is courtesy of an anonymous Abreview
    Anti-MMP2 antibody [4D3] (ab2462) at 1/5000 dilution + Rabbit aorta tissue lysate at 40 µg

    Secondary
    HRP-conjugate Sheep anti-mouse IgG polyclonal

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 72 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-MMP2 antibody [4D3] (ab2462)
    Immunocytochemistry/ Immunofluorescence - Anti-MMP2 antibody [4D3] (ab2462)Image from Kean MJ et al, J Cell Sci. 2009 Nov 15;122(Pt 22):4089-98, Fig 1. DOI 10.1242/?jcs.052761
    ab2462 staining MMP2 in HT-1080 cells by Immunocytochemistry/ Immunofluorescence.
    Cells were grown on glass coverslips, treated with PMA and then fixed with 4% (w/v) paraformaldehyde in phosphate buffered saline (PBS). Samples were then permeabilized with 0.1% Triton X-100 in PBS and blocked with 5% (w/v) skimmed milk powder in PBS before staining with ab2462 and the appropriate secondary antibody, followed by washing and mounting.

    Scale bar: 10 µm.

References

This product has been referenced in:
  • Liang X  et al. Reactive oxygen species mediated oxidative stress links diabetes and atrial fibrillation. Mol Med Rep 17:4933-4940 (2018). Read more (PubMed: 29393403) »
  • Cao Q  et al. Dickkopf-3 upregulation mediates the cardioprotective effects of curcumin on chronic heart failure. Mol Med Rep 17:7249-7257 (2018). Read more (PubMed: 29568962) »
See all 14 Publications for this product

Publishing research using ab2462? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 14 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MMP2 antibody [4D3]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (skin)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: proteinase K
Permeabilization
No
Specification
skin
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 11 2016

Immunohistochemistry (Frozen sections) abreview for Anti-MMP2 antibody [4D3]

 Inconclusive
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skin)
Permeabilization
No
Specification
skin
Blocking step
human serum in 0.25% casein as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Acetone
Read More

Abcam user community

Verified customer

Submitted May 16 2016

Western blot abreview for Anti-MMP2 antibody [4D3]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Rabbit Tissue lysate - whole (aorta)
Specification
aorta
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Jun 24 2013

Question


The KB cells are human cells.
I have stained the B actin very well and everybody here in my lab use the
same secondary antibody without any problem.
I do not think the problem is in the secondary antibody. I have talked with
some researches from other labs and they related the same problem with MMP
What do you think?

Read More
Answer

Thank you for confirming the quality of the secondary antibody. I can believe that the issue is with the MMP2 antibody but I am surprised that all three antibodies are giving you trouble.

Are the problems other people from other labs reporting about the same MMP antibodies from Abcam? Or just the MMP2 antibody?

If you are interested in trying a different MMP2 antibody, I will be happy to send a free replacement if your order is within the limits of our guarantee. Do you know when the order was placed, and the distributor

Read More

Question

I am using proteins extracted from KB cells using RIPA buffer. I am using 20
ul of antibody in 10 mL of TBST + 5% BSA: = 1/500. I am incubating the first ab
overnight (12 h). I have stained these samples with other ab and it is
working pretty good. Do you have any suggestion ?

Read More
Answer

Thank you for the protocol details.


Do the blots for MMP1 and MMP9 look similar to the MMP2 blot? Can you please tell me the species of KB cells?


I am a little concerned aboutthe quality of the sample, given the multiple bands between 50 and 75, and the smearing at the top of the wells. What is the protein that you are staining successfully? Are you using the same secondary for blots of that protein? Is there any possibility that the secondary antibody is the issue?


I look forward to your reply.

Read More

Question

I am using your Anit-MMP2 antibody (ab2462) and as you can see in the
attached figure, I am having a lot of cross-reactivity. As you specify in
the Instructions, this antibody only showed cross-reactivity with MMP-3.
Do you have any idea about what is happening ? I have used milk and BSA
to block (with milk blocking I have really bad results with a lot of
backgroundo). The attached figure is with BSA blocking buffer. The same
thing is happening with my ab2461 and ab3159.

Read More
Answer

Thank you for bringing this to our attention. Can you please describe your samples, how you prepare them for loading into the gel, how much antibody you use and how long you incubate, and any other details you think may be relevant? Are you confident the secondary antibody is effective and is not the source of the background? Have you stained blots of these samples with any other antibodies, and, if so, were the results as you expected?

Read More

Question

Phone call requesting information on the concentration of the Anti-MMP2 antibody [4D3] (ab2462).

Read More
Answer

Thank you for your enquiry.



Thank you for contacting us and sorry for the delay in getting back to you.



As discussed over the phone, unfortunately I can now confirm that the concentration of the Anti-MMP2 antibody [4D3] (ab2462) has not as yet been determined.



The concentration of unpurified antibodies, such as this one sold as tissue culture supernatant, will not have a concentration determined. Antibody concentration is usually determined by protein assay, and tissue culture supernatant will contain a lot of other proteins other than IgG, which means the antibody quantification would not be accurate.

I can however confirm that for tissue culture supernatant, concentration of antibody is known to very between 1 - 3 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question

Received vial of ab2462 which should contain 1ml but cap was loose during shipment and much of the product leaked out of the vial.

Read More
Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab2462.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Question

What is the recommended dilution for using this tissue culture supernatant for IHC?

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Answer

Thank you for your enquiry. We suggest a concentration of 1:20, however we also recommend a pilot experiment using a serial dilution with a range from 1:5 to 1:300 to determine the optimal concentration for the tissue you are staining. I hope this information helps. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

i have 2 questions regaring mmp2 ab2462. does it react against the pro and the active forms of mmp2. can i use placental tissue as positive control. thanks. Nebras Zeizafoun, M.D Pathology Chief Resident St Luke's Roosevelt Hospital Center Beth Israel Medical Center New York, NY ]]] [technical@abcam.com] 09/28/07 4:05 AM ]]]

Read More
Answer

Thank you for your enquiry. This Ab reacts with both the pro- and cleaved form of MMP2. Placental tissue can be used as a positive control. Should you have any further questions then please do not hesitate to get back in touch with me.

Read More

1-10 of 14 Abreviews or Q&A

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