Anti-MMP9 antibody (ab38898)
Key features and details
- Rabbit polyclonal to MMP9
- Suitable for: IHC-Fr, WB
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG
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Overview
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Product name
Anti-MMP9 antibody
See all MMP9 primary antibodies -
Description
Rabbit polyclonal to MMP9 -
Host species
Rabbit -
Specificity
The antibody binds to MMP-9, but does not cross react with the other MMP family members (MMP-1, MMP-2, MMP-3).
In our hands, no signal or a very weak signal was observed in WB in native human samples (BLAST of full length mouse protein sequence showed 72% homology with the Human MMP9 sequence).
Treatment increasing the expression of MMP-9 is recommended when using this antibody.
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Tested applications
Suitable for: IHC-Fr, WBmore details -
Species reactivity
Reacts with: Mouse, Human, Recombinant fragment -
Immunogen
Full length protein corresponding to Mouse MMP9.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 50% Glycerol -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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-Fr | (3) |
1/1000.
(see Abreview submitted by Greg Gibson) We recommend using Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) secondary antibody |
| WB | (20) |
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.
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| Notes |
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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. |
Target
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Function
May 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 specificity
Produced by normal alveolar macrophages and granulocytes. -
Involvement in disease
Intervertebral disc disease
Metaphyseal anadysplasia 2 -
Sequence similarities
Belongs to the peptidase M10A family.
Contains 3 fibronectin type-II domains.
Contains 4 hemopexin repeats. -
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
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 localization
Secreted, extracellular space, extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 4318 Human
- Entrez Gene: 17395 Mouse
- Omim: 120361 Human
- SwissProt: P14780 Human
- SwissProt: P41245 Mouse
- Unigene: 297413 Human
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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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Immunohistochemistry (Frozen sections) - Anti-MMP9 antibody (ab38898)Fransén-Pettersson N. et al PLoS One. 2016 Jul 21;11(7):e0159850. doi: 10.1371/journal.pone.0159850. eCollection 2016. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunofluorescence staining showing MMP9 (green) and DAPI (blue) in 12 weeks old N-IF and 24αβNOD control mouse livers. Scale bars are 100 μm.
Immunohistochemical staining was performed on liver biopsies fixed in 4% paraformaldehyde and embedded in OCT. The frozen tissues were cut in 5 μm thick sections and stained using primary antibody against matrix metallopeptidase 9 (MMP9) (1:300, Abcam, ab38898). The nuclei were visualized with DAPI. The sections where analyzed using confocal microscopy.
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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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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.
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Western blot - Anti-MMP9 antibody (ab38898)All lanes : Anti-MMP9 antibody (ab38898) at 2 µg/ml
Lane 1 :Native 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.
Datasheets and documents
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SDS download
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Datasheet download
References (602)
ab38898 has been referenced in 602 publications.
- Qin T et al. Prediction of the mechanisms of action of Shenkang in chronic kidney disease: A network pharmacology study and experimental validation. J Ethnopharmacol 246:112128 (2020). PubMed: 31386888
- Chen X et al. Exosome-mediated transfer of miR-93-5p from cancer-associated fibroblasts confer radioresistance in colorectal cancer cells by downregulating FOXA1 and upregulating TGFB3. J Exp Clin Cancer Res 39:65 (2020). PubMed: 32293494
- Liu MB et al. Icariside II attenuates cerebral ischemia/reperfusion-induced blood-brain barrier dysfunction in rats via regulating the balance of MMP9/TIMP1. Acta Pharmacol Sin N/A:N/A (2020). PubMed: 32488170
- Kim ML et al. Statins Suppress TGF-ß2-Mediated MMP-2 and MMP-9 Expression and Activation Through RhoA/ROCK Inhibition in Astrocytes of the Human Optic Nerve Head. Invest Ophthalmol Vis Sci 61:29 (2020). PubMed: 32421147
- Huang X et al. Extract of Stellera Chamaejasme L. Inhibits the Progression of Hepatocellular Carcinoma by Regulating miR-134-5p and JAK1/STAT3 Pathway. Cancer Biother Radiopharm N/A:N/A (2020). PubMed: 32486841
Customer reviews and Q&As
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