Revision 1
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, IF-IC, FC-FP

REACTIVITY:

H

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
Immunofluorescence (Immunocytochemistry) 1:100
Flow Cytometry (Fixed/Permeabilized) 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.

Specificity / Sensitivity

Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb recognizes endogenous levels of Vimentin protein only when phosphorylated at Ser83.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser83 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 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).
CDK1 phosphorylates vimentin at Ser56 during mitosis, providing 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).

  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.
  12. Yamaguchi, T. et al. (2005) J Cell Biol 171, 431-6.
  13. Oguri, T. et al. (2006) Genes Cells 11, 531-40.

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 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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 1
#12569

Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb

Western Blotting Image 1: Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa cells, untreated (-), or treated with either hydroxyurea (4 mM, G1/S arrested; +), or Paclitaxel #9807 (100 nM, G2/M arrested; +) for 20 hr, using Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb (upper) and Vimentin (D21H3) XP® Rabbit mAb #5741 (lower).
Immunofluorescence Image 1: Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells using Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb Expand Image
Flow cytometric analysis of untreated Jurkat cells, using Phospho-Vimentin (Ser83) (D5A2D) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 (DNA content). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.