Actin, pan

Actin is one of the most abundant and highly conserved proteins of eukaryotes. Mammalian actins are the product of six different genes with differing distribution patterns in cell types and in tissues. The molecular weight of all six proteins is 42kD, 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, they can be used 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 much in expression as a result of experimental manipulations. Thus, quantitation of the actin band on the western is used as a standard against the band density of other proteins. Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is another "house keeping" protein used in this way (see our popular GAPDH antibody 031365). The actin isotypes were originally classified as a, b and g forms based on their charges which resulted in different mobilities in the isoelectric focusing dimension as seen on 2-dimensional SDS-PAGE. Subsequently, the a spot was found to potentially contain three actin gene products, a-skeletal actin, a-vascular smooth muscle actin, and a-cardiac muscle. The b spot contained a single protein called simply b-actin, while the g spot may contain both g-1 actin and g-2, which are enteric and smooth muscle actins. The six mammalian gene products are very similar as shown by the sequence alignment of the human proteins, which can be downloaded from here. The mostly closely related actin isotypes are 97% identical, while the most distantly related are 94% identical, with most of the variability at the N-terminus (1). With this level of identity antibodies to any one protein may recognize all six isotypes, as is the case with this antibody (see here). Since our antibody binds all six isotypes very strongly on western blots, it is a very effective blotting standard which can work on any cell type or tissue extract. It also works in immunocytochemical experiments, binding stongly and cleanly to filopodia, membrane ruffles and stress fibers, all known to be rich in actin. The HGNC names for the six actin isotypes are ACTA1, ACTA2, ACTC1, ACTB, ACTG1 and ACTG2.\n\nApplications:\nSuitable for use in Immunofluorescence and Western Blot. Other applications not tested.\n\nRecommended Dilution:\nImmunofluorescence: 1:500-1:1000\nWestern Blot: 1:5000-1:10,000; look for a major band at 42kD.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

031348

Size

100ul

Applications

IF, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Bov, Prc

Form

Supplied as a liquid.

P Type

Mab

Purity

Purified by affinity chromatography.

References

1. 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

Additional Info

Recognizes actin. It binds to all six isotypes of mammalian actin. Shows reactivity with human, bovine, porcine, rat and mouse tissues and tissue extracts, and works well on 3T3, HEK293, HeLa and other common cell types grown in tissue culture.