Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Vimentin Antibody Sampler Kit #9775

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Vimentin (Ser56) Antibody #3877 40 µl W IF-IC H M R Mk 57 Rabbit
Phospho-Vimentin (Ser83) Antibody #3878 40 µl W H M R Mk 57 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Vimentin (D21H3) XP® Rabbit mAb #5741 40 µl W IHC-P IF-IC F H M R Mk 57 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

The antibodies in the Vimentin Antibody Sampler Kit detect endogenous levels of total vimetin protein, vimentin only when phosphorylated at Ser56, and when phosporylated at Ser82, respectively.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types, hydroxyurea-treated (4 mM) (G1/S) or paclitaxel-treated (100 nM) (G2/M) for 20 hours, using Phospho-Vimentin (Ser56) Antibody #3877 (upper). β-Actin Antibody #4967 was used as a loading control (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or phosphorylated in vitro by PLK, using Phospho-Vimentin (Ser82) Antibody #3878 (upper). β-Actin Antibody #4967 (lower) was used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Vimentin (D21H3) XP® Rabbit mAb #5741.


Description

The Vimentin Anitbody Sampler Kit provides an economical means to detect total levels of vimentin, vimentin phosphorylated at Ser56, and vimentin phosphorylated at Ser82. The kit contains enough primary and secondary antibody to perform four western mini-blots experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser56, or Ser82 of human vimentin, respectively. Antibodies are purified by Protein A and peptide affinity chromatography. 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).

  1. Eng, L.F. et al. (2000) Neurochem. Res. 25, 1439-1451.
  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, 133-141.
  5. Tang, D.D. et al. (2005) Biochem. J. 388, 773-783.
  6. Fomina, I.G. et al. (1990) Klin. Med. (Mosk.) 68, 125-127.
  7. Nieminen, M. et al. (2006) Nat. Cell Biol. 8, 156-162.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products