Revision 11
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IHC-Bond, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Hm Mk

SENSITIVITY:

Endogenous

MW (kDa):

57

Source/Isotype:

Rabbit IgG

UniProt ID:

#P08670

Entrez-Gene Id:

7431

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
IHC Leica Bond 1:200 - 1:800
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Frozen) 1:50 - 1:100
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #46173.

Specificity / Sensitivity

Vimentin (D21H3) XP® Rabbit mAb detects endogenous levels of total vimentin protein.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg45 of human vimentin protein.

Background

The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are distinguished by their cell-specific expression: cytokeratins (epithelial cells), glial fibrillary acidic protein (GFAP) (glial cells), desmin (skeletal, visceral, and certain vascular smooth muscle cells), vimentin (mesenchyme origin), and neurofilaments (neurons). GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). Research studies have shown that vimentin is present in sarcomas, but not carcinomas, and its expression is examined in conjunction with that of other markers to distinguish between the two (3). Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli help to coordinate various signaling pathways (4). Phosphorylation of vimentin at Ser56 in smooth muscle cells regulates the structural arrangement of vimentin filaments in response to serotonin (5,6). Remodeling of vimentin and other intermediate filaments is important during lymphocyte adhesion and migration through the endothelium (7).

During mitosis, CDK1 phosphorylates vimentin at Ser56. This phosphorylation provides a PLK binding site for vimentin-PLK interaction. PLK further phosphorylates vimentin at Ser83, which might serve as a memory phosphorylation site and play a regulatory role in vimentin filament disassembly (8,9). Additionally, studies using various soft-tissue sarcoma cells have shown that phosphorylation of vimentin at Ser39 by Akt1 enhances cell migration and survival, suggesting that vimentin could be a potential target for soft-tissue sarcoma targeted therapy (10,11).

  1. Eng, L.F. et al. (2000) Neurochem Res 25, 1439-51.
  2. Goebel, H.H. et al. (1987) Acta Histochem Suppl 34, 81-93.
  3. Leader, M. et al. (1987) Histopathology 11, 63-72.
  4. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
  5. Tang, D.D. et al. (2005) Biochem J 388, 773-83.
  6. Fomina, I.G. et al. (1990) Klin Med (Mosk) 68, 125-7.
  7. Nieminen, M. et al. (2006) Nat Cell Biol 8, 156-62.
  8. Yamaguchi, T. et al. (2005) J Cell Biol 171, 431-6.
  9. Oguri, T. et al. (2006) Genes Cells 11, 531-40.
  10. Zhu, Q.S. et al. (2011) Oncogene 30, 457-70.
  11. Xue, G. and Hemmings, B.A. (2013) J Natl Cancer Inst 105, 393-404.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 11
#5741

Vimentin (D21H3) XP® Rabbit mAb

Western Blotting Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mA 对不同细胞系提取物进行蛋白质印迹分析。
Western Blotting Image 2: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb #5741(上图)或 β-Actin (13E5) Rabbit mAb #4970(下图)对对照的 HeLa 细胞(泳道 1)或波形蛋白敲除型 HeLa 细胞(泳道 2)的提取物进行蛋白质印迹分析。波形蛋白敲除型 HeLa 细胞中没有信号,这证实了抗体对波形蛋白的特异性。
Western Blotting Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP ® Rabbit mAb #5741 对 HeLa 细胞的裂解物 (1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunohistochemistry Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的人鳞状细胞肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 2: Vimentin (D21H3) XP® Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的人子宫内膜样腺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 3: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的大鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 4: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的恒河猴肾脏进行免疫组织化学分析。
Immunohistochemistry Image 5: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的叙利亚仓鼠小肠进行免疫组织化学分析。
Immunohistochemistry Image 6: Vimentin (D21H3) XP® Rabbit mAb Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 Vimentin (D21H3) XP® Rabbit mAb 对石蜡包埋的人扁桃体进行免疫组织化学分析。
Immunofluorescence Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 2: Vimentin (D21H3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse colon labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 3: Vimentin (D21H3) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with Vimentin (D21H3) XP® Rabbit mAb (left, green) and co-labeled with F4/80 (BM8.1) Rat mAb (right, red), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
使用 Vimentin (D21H3) Rabbit mAb(绿色)对 SNB19 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Vimentin (D21H3) XP® Rabbit mAb Expand Image
Flow cytometric analysis of MCF7 cells (blue, negative) and HeLa cells (green, positive) using Vimentin (D21H3) XP® Rabbit mAb(solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。