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Overview
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Product nameAnti-MMP9 antibody
See all MMP9 primary antibodies -
DescriptionRabbit polyclonal to MMP9
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Host speciesRabbit
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SpecificityThe antibody binds to Gelatinase-B, but does not cross react with the other MMP family members (MMP-1, MMP-2, MMP-3). In our hands, we observe a weaker signal in WB in human samples compared to mouse samples (BLAST of full length mouse protein sequence showed 72% homology with the Human MMP9 sequence).
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Tested applicationsSuitable for: IHC-P, IHC-Fr, WB, IP, ELISA, ICC/IF, ICC, IHC-FoFrmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human
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Immunogen
Full length protein corresponding to Mouse MMP9.
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Positive control
- HL60 cell lysate. U937 cell lysate. HT1080 cell lysate. Raw 264.7 cell lysate (LPS treated)
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C.
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Storage bufferPreservative: 0.05% Sodium azide
Constituent: 50% Glycerol -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Positive Controls
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Related Products
Applications
Our Abpromise guarantee covers the use of ab38898 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 |
|---|---|---|
| IHC-P | 1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| IHC-Fr | 1/1000. (see Abreview submitted by Greg Gibson) We recommend using Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) secondary antibody |
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| WB | 1/1000. Detects a band of approximately 92 kDa. When using colorimetric substrates such as BCIP/NBT use at a 1/5000 dilution (for chemiluminescent substrates). Detects a band of approximately 92-95 kDa (pro-form) and 82kDa (active form) (Human samples). Mouse MMP9 is larger, and on SDS PAGE gels runs about 102-105 kDa. Dilution optimised using Chromogenic detection. | |
| IP | Use at an assay dependent concentration. | |
| ELISA | Use at an assay dependent concentration. | |
| ICC/IF | 1/500. | |
| ICC | Use at an assay dependent concentration. | |
| IHC-FoFr | Use at an assay dependent concentration. PubMed: 19295156 |
Target
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FunctionMay play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly-
-Leu bond. Cleaves type IV and type V collagen into large C-terminal three quarter fragments and shorter N-terminal one quarter fragments. Degrades fibronectin but not laminin or Pz-peptide. -
Tissue specificityProduced by normal alveolar macrophages and granulocytes.
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Involvement in diseaseIntervertebral disc disease
Metaphyseal anadysplasia 2 -
Sequence similaritiesBelongs to the peptidase M10A family.
Contains 3 fibronectin type-II domains.
Contains 4 hemopexin repeats. -
DomainThe 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.
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Post-translational
modificationsProcessing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9.
N- and O-glycosylated. -
Cellular localizationSecreted, extracellular space, extracellular matrix.
- Information by UniProt
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Database links
- Entrez Gene: 403885 Dog
- Entrez Gene: 4318 Human
- Entrez Gene: 17395 Mouse
- Entrez Gene: 81687 Rat
- Omim: 120361 Human
- SwissProt: O18733 Dog
- SwissProt: P14780 Human
- SwissProt: P41245 Mouse
see all -
Alternative names
- 82 kDa matrix metalloproteinase-9 antibody
- 92 kDa gelatinase antibody
- 92 kDa type IV collagenase antibody
see all
Images
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Western blot - Anti-MMP9 antibody (ab38898)All lanes : Anti-MMP9 antibody (ab38898) at 2 µg/ml
Lane 1 :Natural human MMP9 protein (Proenzyme, monomer) (ab157344)
Lane 2 :Recombinant Mouse MMP9 protein (ab39309)
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Infrared labelled goat anti-rabbit (green) at 1/20000 dilution
Performed under reducing conditions.This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with anti-MMP9 antibody (ab38898; 2 microgram per mL) overnight at 4°C. Antibody binding was detected using infrared labelled goat anti-rabbit (green) antibody (diluted 1:20000) for 1 hour at room temperature before imaging.
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Western blot - Anti-MMP9 antibody (ab38898)Anti-MMP9 antibody (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. -
Western blot - Anti-MMP9 antibody (ab38898)All lanes : Anti-MMP9 antibody (ab38898) at 5 µg/ml
Lane 1 : Recombinant Human MMP9, His tagged (ab82955) at 0.1 µg
Lane 2 : U937 whole cell lysate at 100 µg
Lane 3 : U937 whole cell lysate - treated with PMA and Brefeldin (24 hour treatment) at 100 µg
Lane 4 : Raw 264.7 (Mouse) whole cell lysate at 100 µg
Lane 5 : Raw 264.7 (Mouse) whole cell lysate - treated with LPS (6 hour treatment, 1ug/mL) at 100 µg
Performed under reducing conditions.ab38898 detects recombinant Human MMP9 running at ~85 kDa, and endogenous full-length MMP9 in LPS-stimulated cells at ~100 kDa. This antibody also detects a band at 90 kDa in U937 PMA-treated cells.
ab38898 was incubated at 5 ug/mL and ab8245 (loading control to GAPDH) was diluted at 0.1 ug/mL and both were incubated for 48 hours at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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Western blot - Anti-MMP9 antibody (ab38898)This image is courtesy of an anonymous AbreviewAll lanes : Anti-MMP9 antibody (ab38898) at 1/1000 dilution
Lanes 1-2 : Human lung tissue lysate at 100 µg with 10% Milk for 1 hour at room temperature
Lanes 3-4 : MMP-9 KO mice tissue lysates with 10% Milk for 1 hour at room temperature
Secondary
All lanes : 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 -
Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody (ab38898)Image courtesy of an anonymous Abreviewab38898 staining MMP9 (red) in Mouse Neutrophils and Monocytes cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 2%BSA + 0.2% tritonX100 in PBS. Samples were incubated with primary antibody (1/200 in 2%BSA + 0.2% tritonX100 in PBS) for 25 minutes at 23°C. An Alexa Fluor® 568-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. DAPI is stained blue
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody (ab38898)This image is courtesy of an anonymous Abreviewab38898 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody (ab38898)Image from Jung IH et al., PLoS One. 2011;6(12):e27941. Epub 2011 Dec 2. Fig 7.; doi:10.1371/journal.pone.0027941; December 2, 2011, PLoS ONE 6(12): e27941.ab38898 staining MMP9 in 6 month-old transgenic zebrafish pancreas (Ihha overexpression) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Sections were incubated with primary antibody (1/500) and HRP-conjugated secondary antibody colored using DAB solution. Slides were counterstained with hematoxylin. -
Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody (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. Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) was used as the secondary antibody. In the image: red staining = MMP9, blue staining = nuclei, green = gelatinase activity.
Protocols
References
This product has been referenced in:
- Narumi K et al. (Pro)renin receptor is involved in mesangial fibrosis and matrix expansion. Sci Rep 8:16 (2018). Read more (PubMed: 29311647) »
- Yu R et al. miR-377 targets CUL4A and regulates metastatic capability in ovarian cancer. Int J Mol Med 41:3147-3156 (2018). Read more (PubMed: 29512715) »
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