Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

β-Actin (13E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) #8844

Applications Reactivity Sensitivity Isotype
IF-IC F H M R Mk B Pg (C) (Dg) (Hr) Endogenous Rabbit IgG

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

Protocols

* Product-specific protocol.

Specificity / Sensitivity

β-Actin (13E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) detects endogenous levels of total β-actin protein. This antibody may cross-react with γ-actin (cytoplasmic isoform). It does not cross-react with α-skeletal, α-cardiac, α-vascular smooth, or γ-enteric smooth muscle isoforms.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human β-actin protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using β-Actin (13E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS-7 cells using β-Actin (13E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human and monkey cells, respectively. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated β-Actin (13E5) Rabbit mAb #4970.

Background

Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle β- and γ-actin, also known as cytoplasmic actin, are predominantly expressed in nonmuscle cells, controlling cell structure and motility (1). α-cardiac and α-skeletal actin are expressed in striated cardiac and skeletal muscles, respectively; two smooth muscle actins, α- and γ-actin, are found primarily in vascular smooth muscle and enteric smooth muscle, respectively. These actin isoforms regulate the contractile potential of muscle cells (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (2). The ARP2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (2). Research studies have shown that actin is hyperphosphorylated in primary breast tumors (3). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle, as shown in research studies (4-6). Actin cleavage by caspase-3 may accelerate ubiquitin/proteasome-dependent muscle proteolysis (6).

  1. Herman, I.M. (1993) Curr. Opin. Cell Biol. 5, 48-55.
  2. Condeelis, J. (2001) Trends Cell Biol. 11, 288-293.
  3. Lim, Y.P. et al. (2004) Clin. Cancer Res. 10, 3980-3987.
  4. Kayalar, C. et al. (1996) Proc. Natl. Acad. Sci. USA. 93, 2234-2238.
  5. Communal, C. et al. (2002) Proc. Natl. Acad. Sci. USA. 99, 6252-6256.
  6. Du, J. et al. (2004) J. Clin. Invest. 113, 115-123.

Application References

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Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and7,429,487) from Epitomics, Inc.

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

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