Recombinant human MMP2 protein (Active) (ab125181)

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Functional Studies - Recombinant human MMP2 protein (Active) (ab125181)

    Key features and details

    • Expression system: HEK 293 cells
    • Purity: > 95% SDS-PAGE
    • Active: Yes
    • Tags: His tag C-Terminus
    • Suitable for: Functional Studies, SDS-PAGE

    Description

    • Product name

      Recombinant human MMP2 protein (Active)
      See all MMP2 proteins and peptides

    • Biological activity

      Recombinant Human MMP-2 protein is a pro form. It needs to be activated with p-aminophenylmercuric acetate (APMA).

      The activity was measured by its ability to cleave fluorogenic peptide substrate, Mca-PLGL-DpaAR-NH2.

      The specific activity is > 1,000 pmoles/min/µg.

       

    • Purity

      > 95 % SDS-PAGE.
      ab125181 is purified by proprietary chromatographic techniques.

    • Expression system

      HEK 293 cells

    • Accession

      P08253

    • Protein length

      Full length protein

    • Animal free

      No

    • Nature

      Recombinant

      • Species

        Human

      • Sequence

        APSPIIKFPG DVAPKTDKEL AVQYLNTFYG CPKESCNLFV LKDTLKKMQK FFGLPQTGDL DQNTIETMRK PRCGNPDVAN YNFFPRKPKW DKNQITYRII GYTPDLDPET VDDAFARAFQ VWSDVTPLRF SRIHDGEADI MINFGRWEHG DGYPFDGKDG LLAHAFAPGT GVGGDSHFDD DELWTLGEGQ VVRVKYGNAD GEYCKFPFLF NGKEYNSCTD TGRSDGFLWC STTYNFEKDG KYGFCPHEAL FTMGGNAEGQ PCKFPFRFQG TSYDSCTTEG RTDGYRWCGT TEDYDRDKKY GFCPETAMST VGGNSEGAPC VFPFTFLGNK YESCTSAGRS DGKMWCATTA NYDDDRKWGF CPDQGYSLFL VAAHEFGHAM GLEHSQDPGA LMAPIYTYTK NFRLSQDDIK GIQELYGASP DIDLGTGPTP TLGPVTPEIC KQDIVFDGIA QIRGEIFFFK DRFIWRTVTP RDKPMGPLLV ATFWPELPEK IDAVYEAPQE EKAVFFAGNE YWIYSASTLE RGYPKPLTSL GLPPDVQRVD AAFNWSKNKK TYIFAGDKFW RYNEVKKKMD PGFPKLIADA WNAIPDNLDA VVDLQGGGHS YFFKGAYYLK LENQSLKSVK FGSIKSDWLG C

      • Predicted molecular weight

        71 kDa including tags

      • Amino acids

        30 to 660

      • Tags

        His tag C-Terminus

      • Additional sequence information

        Full length mature protein, minus the signal peptide.

    Specifications

    Our Abpromise guarantee covers the use of ab125181 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    • Applications

      Functional Studies

      SDS-PAGE

    • Form

      Liquid
    • Concentration information loading...

    Preparation and Storage

    • Stability and Storage

      Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.

      pH: 7.40
      Constituents: 0.05% Brij, 0.32% Tris HCl, 0.87% Sodium chloride

      This product is an active protein and may elicit a biological response in vivo, handle with caution.

    General Info

    • Alternative names

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

    • 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

    Images

    • Functional Studies - Recombinant human MMP2 protein (Active) (ab125181)
      Functional Studies - Recombinant human MMP2 protein (Active) (ab125181)This image is courtesy of an Abreview submitted by Mr Pramod Mahajan
      Detection of gelatinolytic activity of ab125181 MMP2 protein.

      10 ng of ab125181 in 25 µl of 25 mM Tris-glycine buffer (pH 6.8) was mixed with 2x SDS-sample buffer (without ß-mercaptoethanol) and incubated at 37°C for 10 minutes. Samples were loaded on a 10% acrylamide-Tris-glycine gel containing 0.1% gelatin and electrophoresed using Tris-glycine (pH 8.3) with 0.1% SDS. Electrophoresis was performed at 90V for 2 hours. The gel was washed 3 times in 40-50 ml of 2.5% Triton X-100 for 15 minutes. The gel was immersed in 40-50 ml of 50 mM Tris-glycine (pH 7.5), 100 mM NaCl, 4 mM CaCl2 and 0.02% Triton X-100 and incubated at 37°C for 16 hours. For detection of the gelatinolytic activity, the gel was stained with 0.05% Commassie Blue.

      ab125181 produced a prominent gelatinolytic band corresponding to a molecular mass of ~ 68 kDa.

      See Abreview

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab125181? Please let us know so that we can cite the reference in this datasheet.

    ab125181 has not yet been referenced specifically in any publications.

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    Other (Biological activity) Abreview for MMP2 protein (Active)

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    Application
    Other - Do not use

    Review text: Ab 125181 is the purified preparation of the recombinant human matrix metalloproteinase 2 proform (rhMMP-2) consisting of amino acids 30- 660 fused with a His tag at the C-terminus. Biological activity of this preparation has thus far been measured using the colorimetric assay based on its ability to cleave the colorimetric peptide substrate, Mca-PLGL-DpaAR-NH2. We tested the biological activity of rhMMP-2 using gelatin, the natural substrate of this enzyme (1).

    Detection of gelatinolytic activity of the recombinant human (rhMMP2) was performed essentially using minor modifications of methods described in the literature (2, 3). Briefly, about 10 ng of rhMMP-2 in 25 µl of 25 mM Tris-glycine buffer pH 6.8 was mixed with equal volume of 2 x SDS-sample buffer without ß-mercaptoethanol and incubated at 370C for 10 minutes. Samples were loaded on a 10 % acrylamide -Tris-glycine gel containing 0.1% gelatin and electrophoresed using Tris-glycine pH 8.3 containing 0.1% SDS. Electrophoresis was performed at 90 volts for 120 minutes. Immediately after electrophoresis, the gel was washed by gentle shaking and incubation in 40-50 ml of 2.5% Triton X-100 for 15 minutes. Washing step was repeated 3 more times. Subsequently, the gel was immersed in 40-50 ml of 50 mM Tris- Glycine pH 7.5, 100 mM NaCl, 4 mM CaCl2 and 0.02% Triton X-100 and incubated at 370C for 16 hours. For detection of the gelatinolytic activity, the gel was stained with 0.05% Commassie Blue for 1 hour, followed by imaging on the ChemiDoc MP Imaging System.

    As seen from Figure 1, the rhMMP-2 sample produced a prominent geletinolytic band corresponding to a molecular mass of ~ 68 kDa. An additional, faint gelatinolytic band, corresponding to a molecular mass ~ 45 kDa also appears to be present in this preparation. Nonetheless, these data confirm the gelatinolytic activity of rhMMP-2. Furthermore, the C-terminal His-Tag does not interfere with the gelatinolytic activity of the recombinant protein.

    Figure 1. Gelatinolytic activity of rhMMP-2 (Ab 151866)



    References:
    1.
    Snoek-van Beurden, P. and Von den Hoff, J. (2005) BioTechniques 38, 73-83
    2.
    Mannello, F. and Sebastiani M. (2003) Clinical Chem. 49, 1546-50
    3.
    Ratnikov, B. Deryugina, E., and Strongin, A. (2002) Laboratory Investigation, 82, 1583-1590
    Sample: Human Recombinant protein

    Primary antibody (in addition to 'MMP2 protein (Active)')
    Primary antibody: None used

    Secondary antibody
    Secondary antibody: None used

    Mr. Pramod Mahajan

    Verified customer

    Submitted Jan 08 2013

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