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  • Product Name
    Vimentin Antibody
  • Catalog No.
    AF7013
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity:
    Human, Mouse, Rat
  • Prediction:
    Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
  • UniProt
  • Mol.Wt.
    53kDa
  • Concentration
    1mg/ml
  • Browse similar products>>

Product Information

Alternative Names:Expand▼

CTRCT30; Epididymis luminal protein 113; FLJ36605; HEL113; VIM; VIME_HUMAN; Vimentin;

Applications:

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

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

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

Specificity:

Vimentin Antibody detects endogenous levels of total Vimentin.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide.

Immunogen Information

Immunogen:

A synthesized peptide derived from human Vimentin

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 53kDa.
Predicted Mol.Wt.: 54kDa.

Subcellular Location:

Cytoplasm.

Tissue Specificity:

Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.

Description:

VIM is an intermediate filament protein. Intermediate filament proteins are expressed in a tissue-specific manner. Desmin is the subunit specific for muscle and vimentin the subunit specific for mesenchymal tissue.

Sequence:
        10         20         30         40         50
MSTRSVSSSS YRRMFGGPGT ASRPSSSRSY VTTSTRTYSL GSALRPSTSR
60 70 80 90 100
SLYASSPGGV YATRSSAVRL RSSVPGVRLL QDSVDFSLAD AINTEFKNTR
110 120 130 140 150
TNEKVELQEL NDRFANYIDK VRFLEQQNKI LLAELEQLKG QGKSRLGDLY
160 170 180 190 200
EEEMRELRRQ VDQLTNDKAR VEVERDNLAE DIMRLREKLQ EEMLQREEAE
210 220 230 240 250
NTLQSFRQDV DNASLARLDL ERKVESLQEE IAFLKKLHEE EIQELQAQIQ
260 270 280 290 300
EQHVQIDVDV SKPDLTAALR DVRQQYESVA AKNLQEAEEW YKSKFADLSE
310 320 330 340 350
AANRNNDALR QAKQESTEYR RQVQSLTCEV DALKGTNESL ERQMREMEEN
360 370 380 390 400
FAVEAANYQD TIGRLQDEIQ NMKEEMARHL REYQDLLNVK MALDIEIATY
410 420 430 440 450
RKLLEGEESR ISLPLPNFSS LNLRETNLDS LPLVDTHSKR TLLIKTVETR
460
DGQVINETSQ HHDDLE

Background

Function:

Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.

Post-translational Modifications:

Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm (PubMed:21465480). Phosphorylated by STK33 (PubMed:18811945). Phosphorylated on tyrosine residues by SRMS (PubMed:29496907).O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.

Subcellular Location:

Cytoskeleton;Nucleus;

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:

Homopolymer assembled from elementary dimers. Interacts with HCV core protein (PubMed:15846844). Interacts with LGSN and SYNM. Interacts (via rod region) with PLEC (via CH 1 domain) (By similarity). Interacts with SLC6A4 (PubMed:19270731). Interacts with STK33 (PubMed:18811945). Interacts with LARP6 (PubMed:21746880). Interacts with RAB8B (By similarity). Interacts with TOR1A; the interaction associates TOR1A with the cytoskeleton (PubMed:16361107, PubMed:18827015). Interacts with TOR1AIP1 (PubMed:16361107). Interacts with BCAS3 (PubMed:17505058). Interacts with DIAPH1 (PubMed:23325789). Identified in complexes that contain VIM, EZR, AHNAK, BFSP1, BFSP2, ANK2, PLEC, PRX and spectrin (By similarity). Interacts with EPPK1; interaction is dependent of higher-order structure of intermediate filament (PubMed:16923132). Interacts with the non-receptor tyrosine kinase SRMS; the interaction leads to phosphorylation of VIM (PubMed:29496907).

Similarity:

The central alpha-helical coiled-coil IF rod domain mediates elementary homodimerization.The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.Belongs to the intermediate filament family.

Research Fields

Research Fields:

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.
· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

Western blot analysis of extracts from Mouse lung, using Vimentin Antibody. The lane on the left was treated with blocking peptide.
AF7013 at 1/200 staining human lymphoma 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.
IHC analysis of paraffin-embedded human liver tissue at dilution of 1:100 using Vimentin Antibody.
AF7013 staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. Samples were then incubated with primary Ab(AF7013 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37°C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.
AF7013 staining A549 cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.

Reference Citations:

1). Han J et al. YY1 complex promotes Quaking expression via super-enhancer binding during EMT of hepatocellular carcinoma. Cancer Res 2019 Feb 13 (PubMed: 30760518) [IF=8.378]

2). Meng J et al. Twist1 Regulates Vimentin through Cul2 Circular RNA to Promote EMT in Hepatocellular Carcinoma. Cancer Res 2018 Aug 1;78(15):4150-4162 (PubMed: 29844124) [IF=8.378]

3). 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]

4). 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]

5). Xu PP et al. JAM-A overexpression is related to disease progression in diffuse large B-cell lymphoma and downregulated by lenalidomide. Sci Rep 2017 Aug 7;7(1):7433 (PubMed: 28785100) [IF=4.011]

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

Lenalidomide (1μM) inhibited JAM-A-transfected cell invasion (B) and EMT (C).


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). Liu YR et al. Selenium-lentinan inhibits tumor progression by regulating epithelial-mesenchymal transition. Toxicol Appl Pharmacol 2018 Sep 18 (PubMed: 30240696) [IF=3.564]

8). Zhao W et al. The role and molecular mechanism of Trop2 induced epithelial-mesenchymal transition through mediated β-catenin in gastric cancer. Cancer Med 2019 Jan 11 (PubMed: 30632714) [IF=3.357]

9). 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]

10). Wang HF et al. Crocetin inhibits the proliferation, migration and TGF-β2-induced epithelial-mesenchymal transition of retinal pigment epithelial cells. Eur J Pharmacol 2017 Nov 15;815:391-398 (PubMed: 28970011) [IF=3.170]

Application: WB    Species:human;    Sample:ARPE-19

Figure 4. Crocetin inhibits TGF-β2-induced EMT. After 1 h of pretreatment with crocetin, ARPE-19 cells used for EMT assay were stimulated with or without recombinant human TGF-β2 for up to 24 or 48 h. (A) Phase contrast photomicrographs of confluent cultures of cells were captured after treatment for 48 h. Scale bar: 200 μm. (B) Western blot analysis levels of of ZO-1, E-cadherin, Vimentin, α-SMA and the housekeeping protein GAPDH in the lysates of ARPE-19 cells after treatment for 48 h. *P< 0.05, **P< 0.01, ***P< 0.001. The data are presented as the mean ± S.D. (n = 3/group).


11). Du Y et al. Estradiol promotes EMT in endometriosis via MALAT1/miR200s sponge function. Reproduction 2018 Nov 1 (PubMed: 30500775) [IF=3.125]

12). Liu H et al. Autophagy contributes to hypoxia-induced epithelial to mesenchymal transition of endometrial epithelial cells in endometriosis. Biol Reprod 2018 May 31 (PubMed: 29860279)

13). Chen G et al. SOSTDC1 inhibits bone metastasis in non-small cell lung cancer and may serve as a clinical therapeutic target. Int J Mol Med 2018 Oct 10 (PubMed: 30320379)

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). Hou F et al. Yi Ai Fang, a traditional Chinese herbal formula, impacts the vasculogenic mimicry formation of human colorectal cancer through HIF-1α and epithelial mesenchymal transition. BMC Complement Altern Med 2016 Nov 2;16(1):428 (PubMed: 27806701)

Application: WB    Species:human;    Sample:Not available

Fig. 2 YAF influences HIF-1α and EMT expression analysis in HCT-116 cell lines. a. Cells were incubated for 48 h in YAF. The level of HIF-1α, Clau-4, E-cd and VIM mRNA were found. b HCT-116 cells were incubated with YAF for 48 h. The level of HIF-1α, Clau-4, E-cd and VIM protiens were found. *P < 0.05. Results are folden change ± SE of at least three independent experiments

Application: IHC    Species:human;    Sample:Not available


16). Wang X et al. Effects of endometrial stem cell transplantation combined with estrogen in the repair of endometrial injury. Oncol Lett 2018 Jul;16(1):1115-1122 (PubMed: 29963188)

17). Gong J et al. Krüppel‑like factor 4 ameliorates diabetic kidney disease by activating autophagy via the mTOR pathway. Mol Med Rep 2019 Aug 9 (PubMed: 31432191)

18). Duan Z et al. Endothelin-1-induced expression of α-smooth muscle actin in human myometrial fibroblasts. J Obstet Gynaecol Res 2018 Mar;44(3):540-546 (PubMed: 29271089)

19). Duan Z et al. Endothelin-1-induced expression of α-smooth muscle actin in human myometrial fibroblasts. J Obstet Gynaecol Res 2018 Mar;44(3):540-546 (PubMed: 29271089)

20). Ai XY et al. Sesquiterpene binding Gly-Leu-Ser/Lys-"co-adaptation pocket" to inhibit lung cancer cell epithelial-mesenchymal transition. Oncotarget 2017 Jul 26;8(41):70192-70203 (PubMed: 29050271)

Application: IHC    Species:human,mouse;    Sample:Not available

Figure 6: Effect of PTL on p-ERK 2, NF-κB, Snail, and EMT protein levels. Brown or yellow staining was observed in the cytoplasmor nucleus. (A and C) Representative photographs of treated and untreated cells. PTL treatment reduced p-ERK 2, NF-κB, and Snail staining compared with sections obtained from control mice. (B and D) Representative photographs of treated and untreated cells. PTL treatment increased E-cadherin and Occludin staining and reduced Vimentin and N-cadherin staining compared with sections obtained from control mice. Each experiment was performed in triplicate. Results show the means of the three experiments, and the error bars represent standard deviation (*P < 0.05 and **P < 0.01).

Application: IF/ICC    Species:human,mouse;    Sample:Not available


21). Hui-Fang Wang et al. Crocetin inhibits the proliferation, migration and TGF-β 2-induced epithelial-mesenchymal transition of retinal pigment epithelial cells . EUR J PHARMACOL 2017 Nov;815:391-398

22). Yang J et al. miR-200b-containing microvesicles attenuate experimental colitis associated intestinal fibrosis by inhibiting epithelial-mesenchymal transition. J Gastroenterol Hepatol 2017 Dec;32(12):1966-1974 (PubMed: 28370348)

Application: WB    Species:rat;    Sample:Not available

Effects of null-MVs and miR-200b-MVs administration on protein expression in TGFb-Indeced EMT.


23). Li XH et al. Parthenolide attenuated bleomycin-induced pulmonary fibrosis via the NF-κB/Snail signaling pathway. Respir Res 2018 Jun 5;19(1):111 (PubMed: 29871641)

24). Zhou P et al. Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis. J Exp Clin Cancer Res 2017 Sep 2;36(1):115 (PubMed: 28865485)

Application: WB    Species:mouse;    Sample:B16F10

Fig. 5 The expression of p-PKCζ, p-cofilin and COX-2 after combined treatment of J-4 and Celecoxib. (a) Western blotting images of p-cofilin and COX-2 in B16-F10 cells with various treatments for 24 h. (b) Western blotting images of p-cofilin and COX-2 in A375 cells with various treatments for 24 h. (c) Relative mRNA level of PKCζ and COX-2 determined via RT-PCR. (d) The expression of EMT markers, E-Cadherin and Vimentin, and MMP-2/MMP-9 was affected in B16-F10 and A375 cells after various treatments for 24 h. J-4: 25 μM; Celecoxib: 25 μM. * P < 0.05; ** P < 0.01


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Catalog Number :

AF7013-BP
(Blocking peptide available as AF7013-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.

Xenopus
100%
Chicken
100%
Rabbit
100%
Pig
100%
Dog
100%
Horse
100%
Bovine
100%
Zebrafish
73%
Sheep
0%
High similarity Medium similarity Low similarity No similarity
P08670 as Substrate
Site PTM Type Enzyme
S2 Phosphorylation
T3 Phosphorylation
R4 Methylation
S5 Phosphorylation P17252 (PRKCA)
S7 O-Glycosylation
S7 Phosphorylation P17252 (PRKCA) , Q96GD4 (AURKB) , Q02156 (PRKCE) , P17612 (PRKACA)
S8 Phosphorylation P17252 (PRKCA)
S9 Phosphorylation P17252 (PRKCA)
S10 Phosphorylation P17252 (PRKCA)
Y11 Phosphorylation
R12 Methylation
R13 Methylation
T20 Phosphorylation
S22 Phosphorylation
R23 Methylation
S25 Phosphorylation P17252 (PRKCA) , Q96GD4 (AURKB) , P17612 (PRKACA)
S26 Phosphorylation Q13153 (PAK1) , P17252 (PRKCA) , Q13177 (PAK2)
S27 Phosphorylation
R28 Methylation
S29 Phosphorylation
Y30 Phosphorylation
T32 Phosphorylation
T33 O-Glycosylation
T33 Phosphorylation
S34 O-Glycosylation
S34 Phosphorylation P17252 (PRKCA)
T35 Phosphorylation
R36 Methylation
T37 Phosphorylation
Y38 Phosphorylation
S39 Phosphorylation P49137 (MAPKAPK2) , Q13177 (PAK2) , P17612 (PRKACA) , O75116 (ROCK2) , P31749 (AKT1) , Q13153 (PAK1) , Q96GD4 (AURKB) , P17252 (PRKCA)
S42 Phosphorylation P17252 (PRKCA)
R45 Methylation
S47 Phosphorylation P17612 (PRKACA) , Q96GD4 (AURKB)
T48 Phosphorylation
S49 Phosphorylation
R50 Methylation
S51 Phosphorylation Q13177 (PAK2) , Q13153 (PAK1) , P49137 (MAPKAPK2)
Y53 Phosphorylation
S55 O-Glycosylation
S55 Phosphorylation P24941 (CDK2) , P06493 (CDK1)
S56 Phosphorylation P78527 (PRKDC) , P06493 (CDK1) , P49137 (MAPKAPK2) , Q13153 (PAK1) , P24941 (CDK2) , Q00535 (CDK5)
Y61 Phosphorylation
T63 Phosphorylation
R64 Methylation
S65 Phosphorylation Q96GD4 (AURKB)
S66 Phosphorylation Q96GD4 (AURKB) , Q13177 (PAK2) , Q13153 (PAK1)
R69 Methylation
R71 Methylation
S72 Phosphorylation P17612 (PRKACA) , Q96GD4 (AURKB) , Q13464 (ROCK1) , O75116 (ROCK2)
S73 Phosphorylation Q13177 (PAK2) , Q96GD4 (AURKB) , P17612 (PRKACA) , Q13153 (PAK1)
S83 Phosphorylation Q13557 (CAMK2D) , Q9UQM7 (CAMK2A) , P49137 (MAPKAPK2) , P53350 (PLK1)
S87 Phosphorylation Q96GD4 (AURKB)
K97 Ubiquitination
T99 Phosphorylation
T101 Phosphorylation
K104 Acetylation
K104 Ubiquitination
R113 Methylation
Y117 Phosphorylation
K120 Acetylation
K120 Methylation
K120 Ubiquitination
K129 Acetylation
K129 Ubiquitination
K139 Acetylation
K139 Ubiquitination
K143 Ubiquitination
S144 Phosphorylation
Y150 Phosphorylation
R158 Methylation
K168 Acetylation
K168 Ubiquitination
R184 Methylation
K188 Ubiquitination
T202 Phosphorylation
S205 Phosphorylation
S214 Phosphorylation
K223 Ubiquitination
S226 Phosphorylation
K235 Acetylation
K235 Ubiquitination
K236 Acetylation
K236 Ubiquitination
S261 Phosphorylation
K262 Ubiquitination
T266 Phosphorylation
Y276 Phosphorylation
S278 Phosphorylation
K282 Acetylation
K282 Ubiquitination
Y291 Phosphorylation
K292 Acetylation
K292 Ubiquitination
K294 Acetylation
K294 Ubiquitination
S299 Phosphorylation
R310 Methylation
K313 Acetylation
K313 Sumoylation
K313 Ubiquitination
S316 Phosphorylation
T317 Phosphorylation
S325 Phosphorylation
T327 Phosphorylation
C328 S-Nitrosylation
K334 Acetylation
K334 Ubiquitination
T336 Phosphorylation
S339 Phosphorylation
Y358 Phosphorylation
T361 Phosphorylation
K373 Acetylation
K373 Ubiquitination
R381 Methylation
Y383 Phosphorylation
Y400 Phosphorylation
R401 Methylation
K402 Acetylation
K402 Sumoylation
K402 Ubiquitination
S409 Phosphorylation
R410 Methylation
S412 Phosphorylation
S419 Phosphorylation
S420 Phosphorylation
T426 Phosphorylation
S430 Phosphorylation P78527 (PRKDC)
T436 Phosphorylation
S438 Phosphorylation
K439 Acetylation
K439 Ubiquitination
T441 Phosphorylation
K445 Acetylation
K445 Methylation
K445 Sumoylation
K445 Ubiquitination
T446 Phosphorylation
T449 Phosphorylation
T458 Phosphorylation P00540 (MOS)
S459 Phosphorylation P78527 (PRKDC) , P53350 (PLK1) , P00540 (MOS)
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.