Beta-actin (human gene and protein abbreviation ACTB/ACTB) is one of six different actin isoforms which have been identified in humans. This is one of the two nonmuscle cytoskeletal actins. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus. Beta-actin has been shown to interact with SPTBN2. In addition, RNA-binding protein Sam68 was found to interact with the mRNA encoding β-actin, which regulates the synaptic formation of the dendritic spines with its cytoskeletal components. Beta-actin has been shown to activate eNOS, thereby increasing NO production. An eight-amino acid residue (326-333) in actin has been shown to mediate the interaction between actin and eNOS. Recurrent mutations in this gene have been associated to cases of diffuse large B-cell lymphoma. Beta actin is often used in Western blotting as a loading control, to normalize total protein amounts and check for eventual protein degradation in the samples. Its transcript is also commonly used as a housekeeping gene standard in qPCR. Its molecular weight is approximately 42 kDa.
背景文献
1. Fujiki R et al. GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459:455-459 (2009).
Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (HA601082) at 1/40,000 dilution.
Lane 1: MCF-7 cell lysate (10 µg/Lane) Lane 2: HepG2 cell lysate (10 µg/Lane) Lane 3: THP-1 cell lysate (10 µg/Lane) Lane 4: PMVEC cell lysate (10 µg/Lane) Lane 5: RH-35 cell lysate (10 µg/Lane) Lane 6: NIH/3T3 cell lysate (10 µg/Lane) Lane 7: RAW264.7 cell lysate (10 µg/Lane) Lane 8: B16F1 cell lysate (10 µg/Lane) Lane 9: MEF cell lysate (10 µg/Lane) Lane 10: Hela cell lysate (10 µg/Lane) Lane 11: F9 cell lysate (10 µg/Lane) Lane 12: PC-12 cell lysate (10 µg/Lane) Lane 13: Mouse lung cell lysate (20 µg/Lane) Lane 14: L6 cell lysate (10 µg/Lane)
Predicted band size: 42 kDa Observed band size: 42 kDa
Exposure time: 30 seconds; 10% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601082) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/100,000 dilution was used for 1 hour at room temperature.
Western blot analysis of beta Actin on HepG2 cell lysates with Mouse anti-beta Actin antibody (HA601082) at different dilutions.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 42 kDa Observed band size: 42 kDa Exposure time: 5 minutes; 10% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601082) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/100,000 dilution was used for 1 hour at room temperature.
Western blot analysis of beta Actin on NIH/3T3 cell lysates with Mouse anti-beta Actin antibody (HA601082) at different dilutions.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 42 kDa Observed band size: 42 kDa
Exposure time: 5 minutes;
10% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601082) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
Western blot analysis of beta Actin on PC-12 cell lysates with Mouse anti-beta Actin antibody (HA601082) at different dilutions.
Lysates/proteins at 10 µg/Lane.
Predicted band size: 42 kDa Observed band size: 42 kDa
Exposure time: 1 minute;
10% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601082) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
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