Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total β-Actin Sandwich ELISA Antibody Pair #7881

Kit Includes Volume Cap Color
Beta-Actin Capture Ab(100X) 0.4 ml Pink
Pan-Actin Detection Ab (100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody 0.04 ml Yellow

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H M R Hm Mk

Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

CST's PathScan® Total β-Actin Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total β-Actin Sandwich ELISA Kit #7880. Capture and detection antibodies (100X stocks) and an HRP-linked secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The β-actin rabbit capture antibody is coated in PBS overnight onto a 96 well microplate. After blocking, cell lysate is added followed by pan-actin mouse detection antibody and HRP-linked, anti-mouse IgG antibody. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of β-actin.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Sandwich ELISA

Sandwich ELISA

Figure 1. The relationship between the protein concentration of the lysate from HeLa cells and the absorbance at 450 nm using the PathScan® Total β-Actin Sandwich ELISA Antibody Pair is shown.

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). It has been reported 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 (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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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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