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
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 Gelatinase-B, 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).
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Tested Applications & Species
See all applications and species dataApplication Species IHC-Fr MouseWB HumanRecombinant fragment
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Immunogen
Full length protein corresponding to Mouse MMP9.
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General notes
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| IHC-Fr |
Mouse
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| WB |
Human
Recombinant fragment
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| 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
- Unigene: 4406 Mouse
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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.
References (508)
ab38898 has been referenced in 508 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
- Huang L et al. Long non-coding RNA NCK1-AS1 promotes the tumorigenesis of glioma through sponging microRNA-138-2-3p and activating the TRIM24/Wnt/ß-catenin axis. J Exp Clin Cancer Res 39:63 (2020). PubMed: 32293515
- Jiang DX et al. Prolyl endopeptidase gene disruption attenuates high fat diet-induced nonalcoholic fatty liver disease in mice by improving hepatic steatosis and inflammation. Ann Transl Med 8:218 (2020). PubMed: 32309365
- Hu H et al. Exosome-Derived miR-486-5p Regulates Cell Cycle, Proliferation and Metastasis in Lung Adenocarcinoma via Targeting NEK2. Front Bioeng Biotechnol 8:259 (2020). PubMed: 32322578
- Despotovic SZ et al. Altered organization of collagen fibers in the uninvolved human colon mucosa 10?cm and 20?cm away from the malignant tumor. Sci Rep 10:6359 (2020). PubMed: 32286443
