| Product Includes | Product # | Volume | Cap Color | Storage Temp |
|---|---|---|---|---|
| β-Actin Capture Rabbit mAb (100X) | 76027 | 400 µl |
|
+4C |
| β-Actin Detection Mouse mAb (100X) | 10367 | 400 µl |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (1000X) | 16736 | 40 µl |
|
-20C |
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Description
CST's PathScan® Total β-Actin Sandwich ELISA Antibody Pair is 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 β-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.
*Antibodies in the kit are custom formulations specific to the kit.
Reagents not supplied
Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809
Cell Lysis Buffer (10X) #9803
TMB Substrate #7004
STOP Solution #7002
Blocking Buffer: 1X PBS/0.5% Tween-20, 1% BSA
96 Well Microplates**
Microplate Reader
** Antibody Pairs have been validated on Corning© 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592).
Notes: Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.
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 ubiquitously expressed, controlling cell structure and motility (1). While all actin isoforms are highly homologous, cytoplasmic β- and γ-actin protein sequences differ by only four biochemically similar amino acids (2). For this reason, antibodies raised to β-actin may cross-react with γ-actin, and vice versa. 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 (3). The ARP2/3 complex stabilizes F-actin fragments and promotes formation of new actin filaments (3). Research studies have shown that actin is hyperphosphorylated in primary breast tumors (4). Cleavage of actin under apoptotic conditions has been observed in vitro and in cardiac and skeletal muscle, as shown in research studies (5-7). Actin cleavage by caspase-3 may accelerate ubiquitin/proteasome-dependent muscle proteolysis (7).
- Herman, I.M. (1993) Curr. Opin. Cell Biol. 5, 48-55.
- Perrin, B.J. and Ervasti, J.M. (2010) Cytoskeleton (Hoboken) 67, 630-4.
- Condeelis, J. (2001) Trends Cell Biol 11, 288-93.
- Lim, Y.P. et al. (2004) Clin Cancer Res 10, 3980-7.
- Kayalar, C. et al. (1996) Proc Natl Acad Sci U S A 93, 2234-8.
- Communal, C. et al. (2002) Proc Natl Acad Sci U S A 99, 6252-6.
- Du, J. et al. (2004) J Clin Invest 113, 115-23.
Background References
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
Limited Uses
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