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Western blot - MMP9 antibody (ab38898)
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Immunohistochemistry (Frozen sections) - MMP9 antibody - Whole molecule (ab38898)
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Western blot - MMP9 antibody - Whole molecule (ab38898)
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Product Name
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MMP9 antibody - Whole molecule
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See all MMP9 antibodies (24)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to MMP9 - Whole molecule
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Immunogen
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Full length MMP-9 native protein. (Mouse)
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Reacts with
(species key)
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Hu, Ms, Dog
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Specificity
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The antibody binds to Gelatinase-B, but does not cross react with the other MMP family members (MMP-1, MMP-2, MMP-3).
We have a range of domain specific antibodies for this target. For a full list please see all MMP9 antibodies
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Tested applications
(see key)
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IHC-FoFr, IHC-Fr, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ELISA: (direct and indirect) Use at an assay dependent dilution.
IHC-Fr: 1/1000 for 1 hr on mouse heart sections (see Abreview submitted by Greg Gibson).
IHC-FoFr: Use at an assay dependent dilution (PMID 19295156).
IHC-P: 1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol (see Abreviews).
IP: Use at an assay dependent dilution.
WB: 1/1000. (when using colorimetric substrates such as BCIP/NBT) - 1/5000 (for chemiluminescent substrates). Detects a band of approximately 92-95 kDa (pro-form) and 82kDa (active form) (Human samples). Mouse MMP-9 is larger, and on SDS PAGE gels runs about 102-105 kDa. Dilution optimised using Chromogenic detection.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
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HL60 cell lysate. U937 cell lysate. HT1080 cell lysate.
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Cellular localization
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Secreted; extracellular matrix
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Research areas
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Cardiovascular >> Atherosclerosis >> Thrombosis >> Other
Cancer >> Tumor biomarkers >> Enzymes >> MMPs
Cell Biology >> Proteolysis / Ubiquitin >> Proteolytic enzymes >> Metalloprotease >> MMPs
Cancer >> Invasion/microenvironment >> ECM >> Extracellular matrix >> MMPs
Cancer >> Invasion/microenvironment >> Angiogenesis >> ECM enzymes >> MMPs
Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> ECM Enzymes >> MMP
Cardiovascular >> Angiogenesis >> Adhesion / ECM >> Matrix Metalloproteinases >> MMP
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Relevance
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All cells within tissues are surrounded by an extracellular matrix (ECM) giving the tissues shape and structure. The ECM is constantly being remodeled and constant communication is maintained between cells through this matrix. Secreted proteins, termed matrix metalloproteinases (MMPs), are involved in the modulation of cell matrix interactions. MMPs are Zn(2+) binding endopeptidases that degrade various components of the ECM. MMPs are enzymes implicated in normal and pathologic tissue remodeling processes, wound healing, angiogenesis, and tumor invasion. These enzymes are very potent when active, and are associated with extracellular space inhibitors called TIMPs (tissue inhibitors of matrix metalloproteinases). TIMPs have been shown to block tumor cell invasion suggesting that they act as metastasis suppressor genes.
MMP9, also known as gelatinase B, is a secreted enzyme which degrades the interstitial collagens, types I, II, and III and is produced by normal alveolar macrophages and granulocytes. The expression of MMP9 increases in Epstein-Barr virus infected lymphoma derived cell lines and may be of significance in typically invasive nasopharyngeal carcinomas. MMP9 is constitutively produced by some tumor cell lines (e.g.: HT1080, HL60, U937) but not by most quiescent cells and tissues. Treatment of cells with the phorbol ester TPA stimulates production of MMP9 in some cell types, but the low protein levels produced (pg/ml) often require concentration of cell culture media to visualize the bands by Western blotting.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: Sodium Azide
Constituents: 50% Glycerol Material safety datasheets (MSDS) for this product:
Glycerol MSDS
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this MMP9 antibody - Whole molecule is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for MMP9 antibody - Whole molecule images, references, products related to ab38898 and other tools.
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MMP9 antibody - Whole molecule images:
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 Western blot - MMP9 antibody (ab38898)
MMP9 antibody - Whole molecule (ab38898) + Human MMP9
Observed band size : 88,92 kDa (why is the actual band size different from the predicted?)
Ab38898 detects a band at 92 Kd (pro-form) and a band at 88 Kd (active form). Mouse MMP9 is slightly larger than human MMP9, and the antibody detects a band at about 105 Kd.
It is recommend to concentrate samples by Gelatin-agarose affinity chromatography prior to Western blot usage. A recommended starting concentration for Western blots is 1:1000 when using colorimetric substrates such as BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentration of antibody may be needed for non-human samples.
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Immunohistochemistry (Frozen sections) - MMP9 antibody - Whole molecule (ab38898)
ab38898 at a 1/1000 dilution staining mouse heart tissue by Immunohistochemistry (Frozen sections). The tissue was removed from a mouse, rinsed in PBS and slowly frozen in supercooled isopentane. 14um sections were made using a cryostat. The sections were acetone fixed and blocked in 2% BSA prior to incubation with the MMP9 antibody. A Cy3 conjugated goat anti-rabbit antibody was used as the secondary antibody. In the image: red staining = MMP9, blue staining = nuclei, green = gelatinase activity.
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Western blot - MMP9 antibody - Whole molecule (ab38898)
All lanes : MMP9 antibody - Whole molecule (ab38898) at 1/1000 dilution
Lane 1 : Human lung tissue lysate at 100 µg with 10% Milk for 1 hour at room temperature
Lane 2 : Human lung tissue lysate at 100 µg with 10% Milk for 1 hour at room temperature
Lane 3 : MMP-9 KO mice tissue lysates with 10% Milk for 1 hour at room temperature
Lane 4 : MMP-9 KO mice tissue lysates with 10% Milk for 1 hour at room temperature
Secondary
HRP-conjugated donkey anti-rabbit polyclonal at 1/1000 dilution
developed using the ECL technique
Performed under reducing conditions.
Specific observed bands 95-100 kDa This image is courtesy of an anonymous Abreview
See Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - MMP9 antibody - Whole molecule (ab38898)
ab38898 staining MMP9 in Mouse Pancreatic carcinoma tissue sections by IHC-P (formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at room temperature. Antigen retrieval was by heat mediation in citric acid (pH6). Samples were incubated with primary antibody (1/100) in 1% Aurion BSA for 12 hours. An HRP-conjugated Donkey polyclonal to rabbit IgG (1/100) was used as secondary antibody. This image is courtesy of an anonymous Abreview
See Abreview
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References for MMP9 antibody - Whole molecule (ab38898)
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This product has been used in: (two most recent references)
Cackowski FC et al. Osteoclasts are important for bone angiogenesis. Blood 115:140-9 (2010). IHC-Fr; Mouse. PubMed: 19887675
El Azreq MA et al. Cytohesin-1 regulates the Arf6-phospholipase D signaling axis in human neutrophils: impact on superoxide anion production and secretion. J Immunol 184:637-49 (2010). WB; Human. PubMed: 20018626
See all 13 publication references for this product.
If you publish research using ab38898 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to MMP9
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