Key features and details
- Rabbit polyclonal to MMP9
- Suitable for: IHC-Fr, WB
- Reacts with: Mouse, Dog, Human
- Isotype: IgG
Product nameAnti-MMP9 antibody
See all MMP9 primary antibodies
DescriptionRabbit polyclonal to MMP9
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).
Tested applicationsSuitable for: IHC-Fr, WBmore details
Species reactivityReacts with: Mouse, Dog, Human
Full length protein corresponding to Mouse MMP9.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C.
Storage bufferPreservative: 0.05% Sodium azide
Constituent: 50% Glycerol
Concentration information loading...
PurityImmunogen affinity purified
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.
(see Abreview submitted by Greg Gibson)
We recommend using Goat Anti-Rabbit IgG H&L (Cy3 ®) preadsorbed (ab6939) secondary antibody
|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.|
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.
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.
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
- 82 kDa matrix metalloproteinase-9 antibody
- 92 kDa gelatinase antibody
- 92 kDa type IV collagenase antibody
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.
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.
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.
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.
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.
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.
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/?-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