PathScan^® Total beta-Actin Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M R Hm Mk

UniProt ID:
#P60709

Entrez-Gene Id:
#60

Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

[email protected]

cellsignal.com

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes	Product #	Quantity	Color	Storage Temp
β-Actin Rabbit mAb Coated Microwells	74040	96 tests		+4C
β-Actin Mouse Detection mAb	23722	1 ea	Green (Lyophilized)	+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)	13304	1 ea	Red (Lyophilized)	+4C
Detection Antibody Diluent	13339	11 ml	Green	+4C
HRP Diluent	13515	11 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
ELISA Sample Diluent	11083	25 ml	Blue	+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The PathScan^® Total β-Actin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of β-actin. A β-actin rabbit antibody has been coated onto the microwells. After incubation with cell lysates, β-actin is captured by the coated antibody. Following extensive washing, a β-actin mouse detection antibody is added to detect the captured β-actin. An anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate (TMB) is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of β-actin.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan^® Total beta-Actin Sandwich ELISA Kit detects endogenous levels of β-actin. As shown in Figure 1, β-actin is readily detected in HeLa cells using this Sandwich ELISA Kit. Total levels of β-actin remain unchanged after IFN-α treatment as shown by western analysis. This Sandwich ELISA Kit does not cross-react with α-smooth muscle actin, α-sarcomeric muscle actin or γ-actin. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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).

Background References

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com