Name: | Mouse monoclonal antibody to all six isotypes of mammalian actin |
Immunogen: | Actin preparation from bovine brain |
HGNC Name: | ACT |
UniProt: | P63267, P60709, P62736, P63261, P68133, P68032 |
Molecular Weight: | 42kDa |
Host: | Mouse |
Isotype: | IgG1 heavy, κ light |
Species Cross-Reactivity: | Human, rat, mouse, cow, pig, horse |
RRID: | AB_2572218 |
Format: | Purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
Applications: | WB, IF/ICC, IHC. |
Recommended Dilutions: | WB: 1:1,000. IF/ICC or IHC: 1:500-1,000. |
Storage: | Store at 4°C for short term and -20°C for long term |
Mouse Monoclonal Antibody to all Actin Isotypes
Cat# MCA-5J11
$120.00 – $800.00
Actin is one of the most abundant and highly conserved proteins of eukaryotes (1-5). Mammalian actins are the product of six distinct but closely related genes with differing distribution patterns in cell types and in tissues. The amino acid sequences of the six human actin gene products are 94-97% identical, so antibodies which bind all six gene products are to be expected. A sequence alignment of the six human actin can be downloaded from here. The molecular weight of all six proteins is ~42kDa, and one or more actins is found in essentially every type of crude cellular and tissue extract. As a result, antibodies to actin are widely used as western blotting standards to verify that the various steps of the western blotting procedure have been performed correctly. In addition, actin is regarded as a “house keeping” protein which is generally not altered in expression as a result of experimental manipulations. Thus, quantification of the actin band on a western blot can be used as a loading control. The actin isotypes were originally classified as α, β and γ since three different actin spots were detected by 2-dimensional SDS-PAGE. Subsequently, the α spot was found to potentially contain three actin gene products, α-skeletal actin, α-vascular smooth muscle actin and α-cardiac muscle actin. The β spot contained a single protein called simply β-actin while the γ spot may contain either γ-1 and/or γ-2 actin, which are enteric and smooth muscle actins respectively. The six mammalian gene products are between 94 and 97% identical, with most of the variability seen at the N-terminus (2). Despite the similarity between the 6 gene products there is some evidence of functional differences between them (6). Actin cycles between monomeric (G-actin) and polymeric microfilaments (F-actin) in a highly regulated manner under the influence of a variety of actin capping and severing proteins the Rho family GTPases, the various Rac, Rho and CDC42 proteins (7).
The MCA-5J11 was made against an actin preparation derived from bovine brain. We have shown that MCA-5J11 binds all six actin gene products, (see supplemental data here), not unexpected given the 94-97% amino acid identity of the 6 proteins. As a result MCA-5J11 will detect all actin proteins present in any mammalian cell or tissue extract, making it a useful and versatile western blotting standard. MCA-5J11 works well for IF, ICC and IHC (see data under “Additional Info” tab). Mouse select image at left for larger view.
Chromogenic immunostaining of a formalin fixed paraffin embedded human brain cortex section with mouse mAb to actin, MCA-5J11, dilution 1:1,000, detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. MCA-5J11 antibody labels the cytoplasm of all cell types tested. This antibody performs well in staining both 4% PFA and standard NBF fixed tissues. Mouse select image for larger view.
GST-fusion proteins encoding all six full length human actin gene products were run out on SDS-PAGE, transferred to nitrocellulose and reacted with MCA-5J11. The MCA-5J11 antibody binds all six proteins strongly, so we conclude that it is a pan specific actin antibody. This is not a surprising finding given the high degree of identity between the six proteins, see below. These constructs can be purchased from Novus Biologicals. The HGNC names for the actin isotypes are two γ gene products. γ-1 actin and γ-2, which are enteric and smooth muscle actins (ACTG1 and ACTG2). The single β-actin is ACTB, and α-skeletal actin, α-vascular smooth muscle actin and α-cardiac muscle are ACTA1, ACTA2, and ACTC1.
The amino acid sequences of the six human actin gene products are 94-97% identical, so antibodies which bind all six gene products are to be expected. A sequence alignment can be downloaded from here.
1. Pollard TD and Cooper JA. Actin, a central player in cell shape and movement. Science 326:1208-12 (2009).
2. Vandekerckhove J. and Weber K. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid. sequence of the amino-terminal tryptic peptide. J. Mol. Biol. 126:783–802 (1978).
3. Dominguez R, Holmes KC. Actin structure and function. Annu. Rev. Biophys. 40:169-86 (2011).
4. Davidson AJ, Wood W. Unravelling the Actin Cytoskeleton: A New Competitive Edge? Trends Cell Biol. 26:569-76 (2016).
5. Blanchoin L, et al. Actin dynamics, architecture, and mechanics in cell motility. Physiol. Rev. 94:235-63 (2014).
6. Skruber et al. Reconsidering an active role for G-actin in cytoskeletal regulation. J. Cell Sci. 10:131 (2017).
7. Hall A. Rho GTPases and the actin cytoskeleton. Science 279:509-14 (1998).
This antibody is fairly new so has not been widely used in peer-reviewed publications. However we will add such citations as and when we discover them.
1. Moradi, M. Differential roles of α-, β- and γ-actin isoforms in regulation of cytoskeletal dynamics and stability during axon elongation and collateral branch formation in motoneurons PhD Thesis University of Würzburg 2017.
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Contact info
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