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  • Product Name
    MMP-9 Antibody
  • Catalog No.
    AF5228
  • Source
    Rabbit
  • Application
    WB,IF/ICC,ELISA,IHC-P
  • Reactivity:
    Human, Mouse, Rat
  • Prediction:
    Pig(82%), Horse(82%)
  • UniProt
  • Mol.Wt.
    78 kDa(active)92-105kd(Pro mmp9)
  • Concentration
    1mg/ml
  • Browse similar products>>

Product Information

Alternative Names:Expand▼

82 kDa matrix metalloproteinase-9; 92 kDa gelatinase; 92 kDa type IV collagenase; CLG 4B; CLG4B; Collagenase Type 4 beta; Collagenase type IV 92 KD; EC 3.4.24.35; Gelatinase 92 KD; Gelatinase B; Gelatinase beta; GelatinaseB; GELB; Macrophage gelatinase; MANDP2; Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase); Matrix Metalloproteinase 9; MMP 9; MMP-9; MMP9; MMP9_HUMAN; Type V collagenase;

Applications:

IF 1:200, IHC-p 1:50-1:200, WB 1:500-1:2000, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(82%), Horse(82%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Specificity:

MMP-9 Antibody detects endogenous levels of total MMP-9.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human MMP-9, corresponding to a region within C-terminal amino acids.

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 78 kDa(active)92-105kd(Pro mmp9).
Predicted Mol.Wt.: 79kDa.

Subcellular Location:

Secreted, extracellular space, extracellular matrix.

Tissue Specificity:

Produced by normal alveolar macrophages and granulocytes.

Description:

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-

Sequence:
        10         20         30         40         50
MSLWQPLVLV LLVLGCCFAA PRQRQSTLVL FPGDLRTNLT DRQLAEEYLY
60 70 80 90 100
RYGYTRVAEM RGESKSLGPA LLLLQKQLSL PETGELDSAT LKAMRTPRCG
110 120 130 140 150
VPDLGRFQTF EGDLKWHHHN ITYWIQNYSE DLPRAVIDDA FARAFALWSA
160 170 180 190 200
VTPLTFTRVY SRDADIVIQF GVAEHGDGYP FDGKDGLLAH AFPPGPGIQG
210 220 230 240 250
DAHFDDDELW SLGKGVVVPT RFGNADGAAC HFPFIFEGRS YSACTTDGRS
260 270 280 290 300
DGLPWCSTTA NYDTDDRFGF CPSERLYTQD GNADGKPCQF PFIFQGQSYS
310 320 330 340 350
ACTTDGRSDG YRWCATTANY DRDKLFGFCP TRADSTVMGG NSAGELCVFP
360 370 380 390 400
FTFLGKEYST CTSEGRGDGR LWCATTSNFD SDKKWGFCPD QGYSLFLVAA
410 420 430 440 450
HEFGHALGLD HSSVPEALMY PMYRFTEGPP LHKDDVNGIR HLYGPRPEPE
460 470 480 490 500
PRPPTTTTPQ PTAPPTVCPT GPPTVHPSER PTAGPTGPPS AGPTGPPTAG
510 520 530 540 550
PSTATTVPLS PVDDACNVNI FDAIAEIGNQ LYLFKDGKYW RFSEGRGSRP
560 570 580 590 600
QGPFLIADKW PALPRKLDSV FEERLSKKLF FFSGRQVWVY TGASVLGPRR
610 620 630 640 650
LDKLGLGADV AQVTGALRSG RGKMLLFSGR RLWRFDVKAQ MVDPRSASEV
660 670 680 690 700
DRMFPGVPLD THDVFQYREK AYFCQDRFYW RVSSRSELNQ VDQVGYVTYD

ILQCPED

Background

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.

Post-translational Modifications:

Processing 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.

Subcellular Location:

Extracellular region or secreted;

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

Exists as monomer or homodimer; disulfide-linked. Exists also as heterodimer with a 25 kDa protein. Macrophages and transformed cell lines produce only the monomeric form. Interacts with ECM1.

Similarity:

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.Belongs to the peptidase M10A family.

Research Fields

Research Fields:

· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Bladder cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Overview > MicroRNAs in cancer.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.
· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > Leukocyte transendothelial migration.(View pathway)
· Organismal Systems > Endocrine system > Relaxin signaling pathway.
· Organismal Systems > Endocrine system > Estrogen signaling pathway.(View pathway)

Western blot analysis of extracts from various samples, using MMP-9 Antibody. Lane 1: B16F10, treated with blocking peptide; Lane 2: B16F10; Lane 3: Mouse muscle. Observed bands: 100 kDa.
Western blot analysis of extracts from various sample,using MMP-9 Antibody. lane1:rat spleen, lane2:mouse brain, lane3:mouse brain with blocking peptide.
AF5228 at 1/200 staining human breast cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF5228 at 1/50 staining human breast cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF5228 staining HepG2 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37°C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.
This image is a courtesy of anonymous review

Reference Citations:

1). Ai XY et al. Phenytoin silver: a new nanocompound for promoting dermal wound healing via comprehensive pharmacological action. Theranostics 2017 Jan 5;7(2):425-435 (PubMed: 28255340) [IF=8.063]

Application: WB    Species:human;    Sample:Not available

Figure 6. PnAg regulates gp130/Jak/Stat3 signaling pathway (A) and (B) NIH-3T3 and HaCat Cells were treated with PnAg at different concentrations and cell viability was tested using MTT analysis. (C) Wound healing assay reflected the effect of PnAg on cell migration. (D) Binding mode of PnAg in the active pocket of gp130. (E) and (F) MMPs activity and expression levels of Stat3, VEGF, TGFB-1, and TGFB1 detected using zymographic and Western blot assays. (G) Diagram of the proposed function of PnAg in wound inflammation and re-epithelialization controls.

Application: IHC    Species:human;    Sample:Not available

Figure 2. PnAg promotes wound healing in SD rats. (A) Photographs of rat skin full-thickness excision wounds on different post-excision days. (B) Change in wound areas of SD rats after treatment; (C) and (D) Expression levels of collagen I, NF-κB, TGF-ß, MMP-2, and MMP-9 in tissues on day 7 and 17 detected by immunohistochemistry. (E) Histogram of protein expression levels in these tissues. (F) and (G) Histomorphological changes in wound tissues stained by Masson trichrome and HE on day 17.


2). Meng J et al. Hsp90β promotes aggressive vasculogenic mimicry via epithelial-mesenchymal transition in hepatocellular carcinoma. Oncogene 2018 Aug 7 (PubMed: 30087438) [IF=6.634]

3). Wang H et al. Oleanolic acid inhibits epithelial-mesenchymal transition of hepatocellular carcinoma by promoting iNOS dimerization. Mol Cancer Ther 2018 Oct 8 (PubMed: 30297361) [IF=4.856]

4). Cao X et al. TRB3 interacts with ERK and JNK and contributes to the proliferation, apoptosis, and migration of lung adenocarcinoma cells. J Cell Physiol 2019 Jun 29 (PubMed: 31256425) [IF=4.522]

5). Yi L et al. Inflammation-mediated SOD-2 upregulation contributes to epithelial-mesenchymal transition and migration of tumor cells in aflatoxin G1-induced lung adenocarcinoma. Sci Rep 2017 Aug 11;7(1):7953 (PubMed: 28801561) [IF=4.011]

Application: WB    Species:human;    Sample:Not available

(d) Te expression of MMP2 and MMP9 at protein level was shown by Western blot. Band intensity is coming from densitometry, and data was shown as mean±SD

Application: IHC    Species:human;    Sample:Not available

(c) Representative immunohistochemical staining of MMP2 and MMP9 in AFG1-induced lung adenocarcinoma.


6). Li H et al. MicroRNA-181a regulates epithelial-mesenchymal transition by targeting PTEN in drug-resistant lung adenocarcinoma cells. Int J Oncol 2015 Oct;47(4):1379-92 (PubMed: 26323677) [IF=3.571]

Application: WB    Species:human;    Sample:A549 cells

Figure 3. A549/DDP and A549/PTX cells showed molecular and morphological changes that were consistent with EMT. (A) microscopy at x200 magnification was used to assess cell morphology. The A549 cells (parental cells) had an epithelioid, rounded cobblestone appearance and there was limited formation of pseudopodia. A549/PTX and A549/DDP cells exhibited a spindle-shaped morphology and an increased formation of pseudopodia, indicating a loss of cell polarity. (B) E-cadherin, β-catenin, vimentin, MMP-2 and MMP-9 which are EMT-related proteins, were assessed in terms of expression levels. EMT-related transcription factors (Snail, Slug, Twist and ZEB1) were measured in A549/PTX and A549/DDP cells using western blot analysis. (C) The expression changes were confirmed at the mRNA level by qRT-PCR. Expression was standardized to the expression of GAPDH and normalized to 1.0 in the parental cells (compared with the parental A549 cells, means ± SEM, n=3, * P<0.05)


7). Wang J et al. Carotid baroreceptor stimulation improves cardiac performance and reverses ventricular remodelling in canines with pacing-induced heart failure. Life Sci 2019 Feb 24 (PubMed: 30811965) [IF=3.448]

8). Tan ZB et al. Rheum palmatum extract exerts anti-hepatocellular carcinoma effects by inhibiting signal transducer and activator of transcription 3 signaling. J Ethnopharmacol 2018 Dec 13 (PubMed: 30553869) [IF=3.414]

9). Bai Y et al. BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo. Mol Carcinog 2017 Mar;56(3):1137-1149 (PubMed: 27770580) [IF=3.411]

Application: WB    Species:human;    Sample:HepG2

Figure 6. Effect of BCL2L10 on its downstream gene expression profiles of human cancer pathway in HepG2 cells. (A) By human cancer pathway PCR array, ectopic expression of BCL2L10 up- or down-regulated several genes related to tumor proliferation, apoptosis, metastasis and angiogenesis. (B) Western blot was performed to confirm the downstream gene expression regulated by BCL2L10 in HepG2 cells. GAPDH was used as an internal control. (C) Schematic diagram of the molecular events for BCL2L10 function as a tumor suppressor through regulating cell cycle, proliferation, apoptosis metastasis and angiogenesis effectors.


10). He Y et al. Quercetin reverses experimental pulmonary arterial hypertension by modulating the TrkA pathway. Exp Cell Res 2015 Nov 15;339(1):122-34 (PubMed: 26476374) [IF=3.329]

Application: WB    Species:rat;    Sample:rat

Fig. 5. Dose-response study of quercetin on the migration of PASMCs in vitro. (A) Photographs of the PASMCs migration through the polycarbonate membrane stained by 0.2% crystal violet in hypoxia and treated with increasing concentrations of quercetin for 24 h. (B) Quantification of the number of cells migrating through the polycarbonate membrane of average of 3 independent experiments. (C) Full-length blots of MMP-2, MMP-9, CXCR4, Integrin α1, β1, and α5 and GAPDH are presented. (D) Results were quantified by densitometry analysis of the bands form (C) and then normalization to GAPDH protein. *Po0.05, **Po0.01 compared with control; #Po0.05, ##Po0.01 compared with hypoxia and quercetin treated PASMCs.


11). Chen Z et al. Lower Expression of Gelsolin in Colon Cancer and Its Diagnostic Value in Colon Cancer Patients. J Cancer 2019 Jan 30;10(5):1288-1296 (PubMed: 30854138) [IF=3.182]

12). Jie L et al. Dencichine ameliorates kidney injury in induced type II diabetic nephropathy via the TGF-β/Smad signalling pathway. Eur J Pharmacol 2017 Oct 5;812:196-205 (PubMed: 28633927) [IF=3.170]

Application: WB    Species:rat;    Sample:Not available

MMP-9 and TIMP-1 western blot analysis (C) was performed on lysates from the kidney cortex, and the band intensity ratios were quantified.

Application: IF/ICC    Species:rat;    Sample:Not available

De increases the ratio of MMP-9 to TIMP-1. The immunofluorescence of MMP-9 and TIMP-1 in vivo (A) (upper panel) and in vitro (B) (lower panel);


13). Liu Y et al. Fucoxanthin Activates Apoptosis via Inhibition of PI3K/Akt/mTOR Pathway and Suppresses Invasion and Migration by Restriction of p38-MMP-2/9 Pathway in Human Glioblastoma Cells. Neurochem Res 2016 Oct;41(10):2728-2751 (PubMed: 27394418)

Application: WB    Species:human;    Sample:u87

d Cell lysates were electrophoresed and MMP- 2/9 and uPA proteins were detected by their respective specific antibodies in indicated concentrations


14). Shao Q et al. MicroRNA-139-5p affects cisplatin sensitivity in human nasopharyngeal carcinoma cells by regulating the epithelial-to-mesenchymal transition. Gene 2018 Apr 30;652:48-58 (PubMed: 29427737)

15). Shi S et al. CRTC2 promotes non-small cell lung cancer A549 migration and invasion in vitro. Thorac Cancer 2018 Jan;9(1):136-141 (PubMed: 29105369)

16). Shi S et al. CRTC2 promotes non-small cell lung cancer A549 migration and invasion in vitro. Thorac Cancer 2018 Jan;9(1):136-141 (PubMed: 29105369)

17). Xie LJ et al. Propofol protects against blood-spinal cord barrier disruption induced by ischemia/reperfusion injury. Neural Regen Res 2017 Jan;12(1):125-132 (PubMed: 28250758)

Application: WB    Species:rabbit;    Sample:Not available

(A) MMP-9 protein expression. MMP-9 expression is expressed as the optical density ratio of MMP-9 to GAPDH.


18). Wu Z et al. MFAP5 promotes tumor progression and bone metastasis by regulating ERK/MMP signaling pathways in breast cancer. Biochem Biophys Res Commun 2018 Apr 6;498(3):495-501 (PubMed: 29526753)

19). Sun C et al. Hsa-miR-326 targets CCND1 and inhibits non-small cell lung cancer development. Oncotarget 2016 Feb 16;7(7):8341-59 (PubMed: 26840018)

Application: WB    Species:Not available;    Sample:Not available

Figure 7: Ectopic expression of miR-326 in A549 and SPC-A-1 cells reduces cell migration and invasion motility. (E) Expression of MMP-7 and MMP-9 protein in A549 and SPC-A-1 cells after transfection.


20). Wang JL et al. Downregulation of castor zinc finger 1 predicts poor prognosis and facilitates hepatocellular carcinoma progression via MAPK/ERK signaling. J Exp Clin Cancer Res 2018 Mar 5;37(1):45 (PubMed: 29506567)

21). et al. Effects and mechanism of siomycin A on the growth and apoptosis of MiaPaCa‑2 cancer cells.

22). et al. High expression of SMARCE1 predicts poor prognosis and promotes cell growth and metastasis in gastric cancer.

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Catalog Number :

AF5228-BP
(Blocking peptide available as AF5228-BP)

Price/Size :

$200/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Horse
82%
Pig
82%
Dog
73%
Bovine
73%
Rabbit
64%
Chicken
0%
Xenopus
0%
Sheep
0%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
P14780 as Substrate
Site PTM Type Enzyme
T37 Phosphorylation
C99 S-Nitrosylation
Y241 Phosphorylation
S628 Phosphorylation
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.