Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Cytoskeletal Signaling

Vimentin (D21H3) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #9856

Applications Reactivity Sensitivity Isotype
IF-IC F H M R Mk Endogenous Rabbit IgG

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Vimentin (D21H3) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of total vimentin protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg45 of human vimentin protein. This antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, using Vimentin (D21H3) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Vimentin (D21H3) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor). Red = Propidium Iodide (fluorescent DNA dye)

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Vimentin (D21H3) XP® Rabbit mAb #5741.

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

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

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