Anti-Vimentin抗体[VI-10] (ab20346)
Key features and details
- Mouse monoclonal [VI-10] to Vimentin
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Chicken, Human
- Isotype: IgM
概述
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产品名称
Anti-Vimentin抗体[VI-10]
参阅全部 Vimentin 一抗 -
描述
小鼠单克隆抗体[VI-10] to Vimentin -
宿主
Mouse -
特异性
The antibody VI-10 reacts with vimentin, a 57 kDa intermediate filament expressed in variety of mesenchymal and mesodermal cell types. -
经测试应用
适用于: WB, IHC-P, Flow Cyt, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Chicken, Human
预测可用于: Pig -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- ICC/IF: HAP1 cells; Mouse dissociated neural precursor cells; Chicken postnatal erythrocytes; RBL rat basophiilioc cells. IHC-P: Human skin tissue. WB: HAP1 cell lysate. Flow Cyt: NIH3T3 cells.
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常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 8.0
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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纯度
Proprietary Purification -
纯化说明
Purified by precipitation and chromatography. -
克隆
单克隆 -
克隆编号
VI-10 -
骨髓瘤
unknown -
同种型
IgM -
轻链类型
unknown -
研究领域
相关产品
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Compatible Secondaries
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Conjugation kits
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab20346于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (4) |
Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
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IHC-P | (4) |
Use a concentration of 1 - 5 µg/ml.
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Flow Cyt | (1) |
Use 1µg for 106 cells.
ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody. |
ICC/IF | (7) |
Use a concentration of 1 µg/ml.
Staining technique: (a) fix cells for 10 min in methanol at -20°C and for 6 min in acetone at -20°C; (b) fix cells directly in methanol for 10 min at -20°C or in acetone for 10 min at -20°C. |
说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 54 kDa. |
IHC-P
Use a concentration of 1 - 5 µg/ml. |
Flow Cyt
Use 1µg for 106 cells. ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 1 µg/ml. Staining technique: (a) fix cells for 10 min in methanol at -20°C and for 6 min in acetone at -20°C; (b) fix cells directly in methanol for 10 min at -20°C or in acetone for 10 min at -20°C. |
靶标
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功能
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.
Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2. -
组织特异性
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. -
疾病相关
Cataract 30 -
序列相似性
Belongs to the intermediate filament family. -
结构域
The central alpha-helical coiled-coil rod region 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. -
翻译后修饰
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. Phosphorylated by STK33.
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. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 420519 Chicken
- Entrez Gene: 7431 Human
- Entrez Gene: 22352 Mouse
- Entrez Gene: 100522394 Pig
- Entrez Gene: 81818 Rat
- Omim: 193060 Human
- SwissProt: P09654 Chicken
- SwissProt: P08670 Human
see all -
形式
Vimentin is found in connective tissue and in the cytoskeleton. -
别名
- CTRCT30 antibody
- Epididymis luminal protein 113 antibody
- FLJ36605 antibody
see all
图片
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All lanes : Anti-Vimentin antibody [VI-10] (ab20346) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : VIM (Vimentin) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 54 kDaLanes 1 - 2: Merged signal (red and green). Green - ab20346 observed at 57 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab20346 was shown to specifically react with Vimentin in wild-type HAP1 cells as signal was lost in VIM (Vimentin) knockout cells. Wild-type and VIM (Vimentin) knockout samples were subjected to SDS-PAGE. Ab20346 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
ab20346 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab20346 at 1μg/ml dilution and ab202272 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human skin tissue labeling Vimentin with ab20346 at 5 ug/ml.
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Immunocytochemical analysis of chicken postnatal erythrocytes labeling Vimentin with ab20346 at 1 ug/ml (green). Cells were counterstained with anti-alpha-tubulin (red) and DAPI (blue).
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ab20346 at a 1/500 dilution staining mouse dissociated neural precursor cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a goat anti-mouse Alexa-Fluor 568 (red stain) ab175473>ab175473). The image also shows a blue nuclear counterstain. Confocal image is shown with 2x zoom detail in top right corner.
This image is courtesy of an Abreview submitted by Randal Moldich on 22 February 2006.
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Human normal skin. Staining is localised to cytoplasm. Left panel: with primary antibody at 1 ug/mL. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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ab20346 used at a 1/1000 dilution in Western Blot.5-20µg of total protein from each sample was loaded.
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Overlay histogram showing NIH3T3 cells stained with ab20346 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab20346, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in NIH3T3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.
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Immunofluorescence staining of RBL rat basophiilioc cell line with anti-Vimentin (VI--10). Nuclei are stained with DAPI (blue).
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (128)
ab20346 被引用在 128 文献中.
- Xue T et al. An oral phenylacrylic acid derivative suppressed hepatic stellate cell activation and ameliorated liver fibrosis by blocking TGF-β1 signalling. Liver Int 43:718-732 (2023). PubMed: 36448910
- Li C et al. Glutaredoxin 1 protects lens epithelial cells from epithelial-mesenchymal transition by preventing casein kinase 1α S-glutathionylation during posterior capsular opacification. Redox Biol 62:102676 (2023). PubMed: 36989576
- Mostert D et al. Methacrylated human recombinant collagen peptide as a hydrogel for manipulating and monitoring stiffness-related cardiac cell behavior. iScience 26:106423 (2023). PubMed: 37035009
- Voss G et al. Functional consequences of A-to-I editing of miR-379 in prostate cancer cells. Sci Rep 13:16602 (2023). PubMed: 37789115
- Grolleman J et al. Environmental stiffness restores mechanical homeostasis in vimentin-depleted cells. Sci Rep 13:18374 (2023). PubMed: 37884575