Product: MMP9 Antibody
Catalog: AF5228
Description: Rabbit polyclonal antibody to MMP9
Application: WB IHC IF/ICC IHC-P
Reactivity: Human, Mouse, Rat
Prediction: Pig, Horse
Mol.Wt.: 78 kDa(active),92-105kd(Pro mmp9); 78kD(Calculated).
Uniprot: P14780
RRID: AB_2837714

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
IF/ICC 1:200, IHC-p 1:50-1:200, WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(82%), Horse(82%)
Clonality:
Polyclonal
Specificity:
MMP9 Antibody detects endogenous levels of total MMP9.
RRID:
AB_2837714
Cite Format: Affinity Biosciences Cat# AF5228, RRID:AB_2837714.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
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.
Alias:

Fold/Unfold

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;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P14780 MMP9_HUMAN:

Detected in neutrophils (at protein level) (PubMed:7683678). 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:
MSLWQPLVLVLLVLGCCFAAPRQRQSTLVLFPGDLRTNLTDRQLAEEYLYRYGYTRVAEMRGESKSLGPALLLLQKQLSLPETGELDSATLKAMRTPRCGVPDLGRFQTFEGDLKWHHHNITYWIQNYSEDLPRAVIDDAFARAFALWSAVTPLTFTRVYSRDADIVIQFGVAEHGDGYPFDGKDGLLAHAFPPGPGIQGDAHFDDDELWSLGKGVVVPTRFGNADGAACHFPFIFEGRSYSACTTDGRSDGLPWCSTTANYDTDDRFGFCPSERLYTQDGNADGKPCQFPFIFQGQSYSACTTDGRSDGYRWCATTANYDRDKLFGFCPTRADSTVMGGNSAGELCVFPFTFLGKEYSTCTSEGRGDGRLWCATTSNFDSDKKWGFCPDQGYSLFLVAAHEFGHALGLDHSSVPEALMYPMYRFTEGPPLHKDDVNGIRHLYGPRPEPEPRPPTTTTPQPTAPPTVCPTGPPTVHPSERPTAGPTGPPSAGPTGPPTAGPSTATTVPLSPVDDACNVNIFDAIAEIGNQLYLFKDGKYWRFSEGRGSRPQGPFLIADKWPALPRKLDSVFEERLSKKLFFFSGRQVWVYTGASVLGPRRLDKLGLGADVAQVTGALRSGRGKMLLFSGRRLWRFDVKAQMVDPRSASEVDRMFPGVPLDTHDVFQYREKAYFCQDRFYWRVSSRSELNQVDQVGYVTYDILQCPED

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
82
Horse
82
Bovine
73
Dog
73
Rabbit
64
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P14780 As Substrate

Site PTM Type Enzyme
T37 Phosphorylation
C99 S-Nitrosylation
Y241 Phosphorylation
S628 Phosphorylation

Research Backgrounds

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.

PTMs:

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:

Secreted>Extracellular space>Extracellular matrix.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Detected in neutrophils (at protein level). Produced by normal alveolar macrophages and granulocytes.

Subunit Structure:

Exists as monomer or homodimer; disulfide-linked. Exists also as heterodimer with LCN2. Macrophages and transformed cell lines produce only the monomeric form. Interacts with ECM1.

(Microbial infection) Interacts with Staphylococcus aureus protein SSL5; this interaction inhibits MMP9 activity.

Family&Domains:

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

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder 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 > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Liao et al. CRSP8 promotes thyroid cancer progression by antagonizing IKKα-induced cell differentiation. Cell Death & Differentiation 2021 Apr;28(4):1347-1363. (PubMed: 33162555) [IF=12.4]

Application: WB    Species: Human    Sample: thyroid cancer cells

Fig. 3 Regulation of CRSP8 on migration, invasion, and apoptosis of thyroid cancer cells. A Cell migration and B cell invasion were analyzed in thyroid cancer cells transfected with CRSP8 specific siRNAs or its overexpression plasmids, and the relative rate of migration was calculated, the number of invading cells was counted. C Western blot analysis of the expression of MMP-9, EMT markers (E-cadherin, N-cadherin, β-catenin, Vimentin) in THJ-29T and FTC-133 cells following CRSP8 silencing or overexpression. THJ-21T and THJ-29T cells were treated with CRSP8 specific siRNAs for 48 h, then D FACS analysis and E acridine orange/ethidium bromide fluorescence staining were performed, and the apoptotic cell ratios were counted respectively. F Western blot analysis of the expression of Bcl-2, Bad, Bim, Bax, cleaved caspase-3, and cleaved caspase-9 proteins in thyroid cancer cells following CRSP8 knockdown. The data represent the mean ± SD of at least three independent experiments (four independent experiments in 3D), and the level of significance was indicated by ***P < 0.001.

2). 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=12.4]

Application: WB    Species: human    Sample:

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:

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.

3). Wang et al. Downregulation of castor zinc finger 1 predicts poor prognosis and facilitates hepatocellular carcinoma progression via MAPK/ERK signaling. Journal of Experimental & Clinical Cancer Research 2018 Mar 5;37(1):45 (PubMed: 29506567) [IF=11.3]

4). Kou L et al. Opsonized nanoparticles target and regulate macrophage polarization for osteoarthritis therapy: A trapping strategy. Journal of Controlled Release 2022 Jul;347:237-255. (PubMed: 35489544) [IF=10.8]

5). Yang Y et al. The NQO1/PKLR axis promotes lymph node metastasis and breast cancer progression by modulating glycolytic reprogramming. CANCER LETTERS 2019 Apr 4 (PubMed: 30954648) [IF=9.7]

6). Lu Q et al. Echinatin inhibits the growth and metastasis of human osteosarcoma cells through Wnt/β-catenin and p38 signaling pathways. Pharmacological Research 2023 Apr 04; (PubMed: 37023991) [IF=9.3]

Application: WB    Species: Human    Sample: OS cells

Fig. 4. Ecn inhibits the migration and invasion of OS cells. (A) The effect of Ecn on the migration of OS cells (Wound healing assay, 100 ×). (B) The effect of Ecn on the migration of OS cells (Transwell assay, 100 ×). (C) The effect of Ecn on the invasion of OS cells (Matrigel-coated Transwell assay, 100 ×). (D) The effect of Ecn on the protein level of MMP2, MMP7, MMP9, Snail, Vimentin, N-Cadherin and E-Cadherin in OS cells (Western blot). ##P 

7). Duan R et al. A De Novo Frameshift Mutation in TNFAIP3 Impairs A20 Deubiquitination Function to Cause Neuropsychiatric Systemic Lupus Erythematosus. JOURNAL OF CLINICAL IMMUNOLOGY 2019 Oct 17 (PubMed: 31625129) [IF=9.1]

8). Luo Y et al. Foxq1 promotes metastasis of nasopharyngeal carcinoma by inducing vasculogenic mimicry via the EGFR signaling pathway. Cell Death & Disease 2021 Apr 19;12(5):411. (PubMed: 33875643) [IF=9.0]

Application: IF/ICC    Species: mouse    Sample: 5–8F cells and CNE1

Fig. 4| Foxq1 promotes the EGFR signaling pathway and VM-related genes through a direct binding to the EGFR promoter. A Immunofluorescence staining of Foxq1,EGFR,VE-cadherin,MMP2, and MMP9 in Foxq1 dysregulated 5–8F, and CNE1; Scale bars represent 50 μm. qRTPCR (above) and western blot (down) were used to monitor the expression of EGFR signaling pathway and VM-related genes in 5–8F, and CNE1 after Foxq1 downregulation (B, C) or overexpression (D, E)

Application: WB    Species: mouse    Sample: 5–8F cells

Fig. 6 |Erlotinib and Nimotuzumab could inhibit Foxq1-induced VM formation and NPC growth and metastasis in vivo.F Statistical results of metastatic nodules of each group; p < 0.001. The expression of related genes in xenograft tumors from each group were monitored by qRT-PCR (G) and western blot (H).

9). Meng J et al. RETRACTED ARTICLE: Hsp90β promotes aggressive vasculogenic mimicry via epithelial–mesenchymal transition in hepatocellular carcinoma. Oncogene 2019 Jan;38(2):228-243 (PubMed: 30087438) [IF=8.0]

10). Meng J et al. Hsp90β promotes aggressive vasculogenic mimicry via epithelial-mesenchymal transition in hepatocellular carcinoma. ONCOGENE 2018 Aug 7 (PubMed: 30087438) [IF=8.0]

Application: IHC    Species: human    Sample: HCC cells

Fig. 1| Hsp90βassociates with vasculogenic mimicry and poor prog-nosis in HCC. e Representative images of Hsp90β, Hsp90α, VE-cadherin,E-cadherin, Vimentin, MMP2, and MMP9 expression in VM/Hsp90βnegative and positive HCC tissue samples. All images are repre-sentative.

Application: WB    Species: human    Sample: PLC-PRF-5 cells

Fig. 5| Hsp90β promotes Twist1 nuclear translocation and binding to VE-cadherin promoter to increase VM-related gene networks. eWestern blot analysis of VM and EMT-related markers, including VE-cadherin, VEGFR1, VEGFR2, E-cadherin, Vimentin, MMP2, and MMP9 in PLC-PRF-5 cells overexpressed Hsp90β or under lack of Twist1.

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