Tested ApplicationsPositive WB detected inrat testis tissue, NCCIT cells,Neuro-2a cells,L02 cells,pig cerebellum tissue,rat brain tissue,mouse ovary tissue,HEK-293 cells,mouse testisPositive IHC detected inmouse ovary tissue, human gliomas tissue,rat brain tissue,mouse brain tissue,human lung cancer 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, mouse brain tissuePlanning an IF experiment? We recommend our CoraLite®488 and CoraLite®594 conjugated versions of this antibody.Recommended dilutionApplicationDilutionWestern Blot (WB)WB : 1:2000-1:10000Immunohistochemistry (IHC)IHC : 1:1000-1:4000Immunofluorescence (IF)IF : 1:50-1:500Sample-dependent, check data in validation data galleryPublished ApplicationsWBSee 20 publications belowIFSee 16 publications belowProduct Information66200-1-Ig targets acetylated Tubulin(Lys40) inWB, IHC, IF, ELISA applications and shows reactivity with human,mouse,rat,dog, pig samples.Tested Reactivityhuman,mouse,rat,dog, pigCited Reactivity bovine, Drosophila, human, mouse, rat, swineHost / IsotypeMouse / IgG1ClassMonoclonalTypeAntibodyImmunogenPeptideFull Nametubulin, alpha 1aCalculated molecular weight 52 kDaObserved molecular weight 55 kDaGenBank accession numberNM_006009Gene symbolTUBA1AGene ID (NCBI)7846ConjugateUnconjugatedFormLiquidPurification MethodProtein G 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 InformationWhat is the molecular weight of acetyl-α-tubulin?The molecular weight of acetylated tubulin is 52 kD.Where does the acetylation of Lys 40 occur?This acetylation occurs inside the microtubule lumen by the α-tubulin acetyltransferase 1 (αTAT1) (PMID: 29207274).How can acetylation be reversed?Acetylation of Lys 40 can be reversed by deacetylase 6 (HDAC6), which is mostly cytoplasmic that also deacetylates Hsp90, and sirtuin 2 (SIRT2), which also mainly cytoplasmic and uses NAD as a coenzyme. Unlike HDAC6, SIRT uses both polymerized and soluble tubulin as substrates. Deacetylases are believed to be more active on soluble tubulin, while acetylases function preferentially on stable polymers (PMIDs: 29207274, 30079247, 19185337).What is the function of acetylation?Acetylation is a conserved post-translational modification of alpha tubulin at Lys 40 during tubulin assembly, and it correlates to increased microtubule stability and intracellular transport (PMIDs: 29207274, 30079247, 20940043). Is acetylation of α-tubulin strictly associated with stable microtubules?Not necessarily, as acetylation can have other effects on microtubule subpopulations (PMID 20940043).Is ac-tubulin found only in cilia?Acetylated-α-tubulin is located in cytoplasmic tubulin as well as in cilia; therefore, it is not strictly region-specific (PMID: 30079247).What are the cellular effects of tubulin acetylation?Microtubule acetylation seems to provide a critical role in neuronal development and function, and while its effect on cancer cells remains unclear, it has been shown that decreased acetylated α-tubulin impairs neuronal cell line migration. The post-translational modification may also help regulate organelle-independent signalling throughout the cell, supporting the notion of a microtubule network serving as a coordinator of cellular signaling (PMIDs: 29207274, 25503560, 20940043, 19185337).ProtocolsProduct Specific ProtocolsWB protocol for acetylated Tubulin(Lys40) antibody 66200-1-IgDownload protocolIHC protocol for acetylated Tubulin(Lys40) antibody 66200-1-IgDownload protocolIF protocol for acetylated Tubulin(Lys40) antibody 66200-1-IgDownload protocolStandard ProtocolsClick here to view our Standard Protocols