Tested ApplicationsPositive WB detected inmulti-cells, Jurkat cells,A549 cells,CHO cells,HeLa cellsPositive IP detected inHeLa cellsPositive IHC detected inhuman kidney tissue,human brain tissue,human colon cancer tissue,human heart tissueNote: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0Positive IF detected inMDCK cells, HepG2 cellsPositive FC detected inHeLa cellsPlanning an IF experiment? We recommend our CoraLite®488 and CoraLite®594 conjugated versions of this antibody.Recommended dilutionApplicationDilutionWestern Blot (WB)WB : 1:5000-1:50000Immunoprecipitation (IP)IP : 0.5-4.0 ug for IP and 1:5000-1:50000 for WBImmunohistochemistry (IHC)IHC : 1:20-1:2000Immunofluorescence (IF)IF : 1:200-1:1500Sample-dependent, check data in validation data galleryPublished ApplicationsWBSee 1247 publications belowIHCSee 1 publications belowIFSee 6 publications belowProduct InformationThe immunogen of 66009-1-Ig is Beta Actin Fusion Protein expressed in E. coli.Tested Reactivityhuman,mouse,rat,hamster,zebrafish, monkeyCited Reactivity broiler chick, chick, chicken, chicks, common carp, common carps, cow, dog, Drosophila, duckHost / IsotypeMouse / IgG2bClassMonoclonalTypeAntibodyImmunogenFusion ProteinFull Nameactin, betaCalculated molecular weight 42 kDaObserved molecular weight 42 kDaGenBank accession numberNM_001101Gene symbolACTBGene ID (NCBI)60ConjugateUnconjugatedFormLiquidPurification MethodProtein A purificationStorage BufferPBS with 0.02% sodium azide and 50% glycerol pH 7.3.Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for-20oC storage.Background InformationActins are highly conserved globular proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. At least six isoforms of actins are known in mammals and other vertebrates. Beta-actin (or β-actin) is a cytoskeletal actin isoform, highly conserved and ubiquitously expressed in eukaryotic organisms, making it a commonly used "housekeeping gene" in many laboratory techniques. It has been widely used as the internal control in RT-PCR and Western Blotting. The isotype of this antibody is IgG2b.What is the molecular weight of beta-actin? 42 kDaWhat is the function of beta-actin?Actins are a key part of the cytoskeleton and are involved in cell motility, structure, and signaling, with beta-actin as part of the contractile apparatus. Beta-actin is located in the cytoplasm of non-muscle cells, particularly where the plasma membrane meets the cytoskeleton (PMID: 1993736) and is found co-distributed with alpha-actin (PMID: 9490838).What are the applications for beta-actin?As it is ubiquitously and constitutively expressed, beta-actin is a useful loading control in techniques such as RT-PCR and Western Blots to measure baseline expression and degradation. Beta-actin can be used as a reference gene to normalize mRNA or protein levels in quantitative studies of expression. Although some evidence suggests variation in tissue types and developing embryos (PMID: 23099774) if expression is stable across conditions, beta-actin is still considered by many to be a suitable gene for normalization.Beyond its role as a housekeeping gene, it has been shown that beta-actin associates with endothelial nitric-oxide synthase (eNOS) and can therefore regulate nitric oxide activity in endothelial cells (PMID: 12734108).What are the post-transcriptional modifications (PTMs) to beta actin?Alterations to actin proteins can modulate their properties and function. While small molecules are known to alter actin, post-translational modifications such as phosphorylation and acetylation can also affect actin (PMID: 23195437). Modifications to actin have been associated with diseases including but not limited to age-related macular degeneration (PMID: 17652714), cardiovascular disease (PMID: 21232667), and Alzheimer"s disease (PMID: 11246152).For murine tissue sample, conjugated antibody (HRP-66009) or rabbit antibody (20536-1-AP) is preferable.ProtocolsProduct Specific ProtocolsWB protocol for Beta Actin antibody 66009-1-IgDownload protocolIHC protocol for Beta Actin antibody 66009-1-IgDownload protocolIF protocol for Beta Actin antibody 66009-1-IgDownload protocolIP protocol for Beta Actin antibody 66009-1-IgDownload protocolFC protocol for Beta Actin antibody 66009-1-IgDownload protocolStandard ProtocolsClick here to view our Standard Protocols