Beta actin

Manufactured by CWBIO

Sourced in China

Beta-actin is a highly conserved cytoskeletal protein that plays a fundamental role in various cellular processes. It is commonly used as a reference gene or loading control in molecular biology and cell biology experiments.

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Mouse anti beta-ACTIN

3 protocols using beta actin

Setdb1 Knockdown Immunoblotting Protocol

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The cells were transfected with Setdb1 siRNA for 48 h. Approximately 30 μg protein was separated by 8–12% SDS-PAGE and transferred to PVDF membranes (Millipore). The membranes were probed using the following primary antibodies: NOX4 (1:500; NB110-58849; Novus), beta-Actin (1:2000; CW0096; CWBIO), SETDB1 (1:1000; 11231-1-AP; Proteintech), E2F1 (1:500; sc-193; Santa Cruz Biotechnology), FOXO4 (1:500; sc-5221; Santa Cruz Biotechnology), Lamin B (1:500; sc-6217; Santa Cruz Biotechnology), JNK (1:500; sc-7345; Santa Cruz Biotechnology), p-JNK (1:400; WL01813; WanleiBio), γH2AX (1:000; 2577; Cell signaling technology), p38 (1:500; sc-7972; Santa Cruz Biotechnology), and p-P38 (1:500; sc-17852-R; Santa Cruz Biotechnology). All were used as the manufacturer’s recommendation. The secondary antibodies were horse radish peroxidase-linked anti-mouse, anti-rabbit, or anti-goat IgG for 2 h at room temperature. The membranes were visualized on a Bio-Rad Chemidoc XRS using a Western Bright ECL Kit (Bio-Rad, Berkeley, CA, United States).

Li X., Chen X., Liu Y., Zhang P., Zheng Y, & Zeng W. (2020). The Histone Methyltransferase SETDB1 Modulates Survival of Spermatogonial Stem/Progenitor Cells Through NADPH Oxidase. Frontiers in Genetics, 11, 997.

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Acetylation analysis of TMV proteins

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Proteins were isolated from
samples that had been inoculated with PBS and TMV solutions at different
time points. We used a commercial antibody against acetylation on
Western blot analysis as previously reported.^{26 (link),45 (link)} The proteins were isolated by sodium dodecyl sulfate–polyacrylamide
gel electrophoresis (SDS–PAGE) and then transferred to a polyvinylidene
fluoride (PVDF) membrane (Immobilon-P, Merck Millipore, United States).
The TMV coat protein (cp) antibody (Agdia, Elkhart, United States),
anti-rabbit secondary antibody (CWBIO, Beijing, China), and beta-actin
(CWBIO, Beijing, China) antibody were used for the analysis. Anti-acetyl-lysine
was used as a primary antibody (Micron Bio, Hangzhou, China) and the
anti-mouse secondary antibody conjugated to HRP (CWBIO, Beijing, China).
The Western blot results were analyzed using ImageJ (v.1.52a, NIH,
Bethesda, USA).

Song L., Zhan H., Wang Y., Lin Z., Li B., Shen L., Jiao Y., Li Y., Wang F, & Yang J. (2022). Cross-Talk of Protein Expression and Lysine Acetylation in Response to TMV Infection in Nicotiana benthamiana. ACS Omega, 7(36), 32496-32511.

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Protein Quantification and Western Blotting

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Total proteins were collected using RIPA buffer (Thermo Scientific, CA, USA) and then quantified for protein quantification by using Pierce™ BCA Protein Quantification Kit (Thermo Scientific) [15 (link)]. Protein (20 μg) was used for electrophoresis loading SDS-PAGE and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% skimmed milk and incubated with primary antibodies, including anti-KIAA1429 (Cell Signaling Technology, 1:1000, #88,358), anti-ROCK2 (Cell Signaling Technology, 1:1000, #47,012), and beta-actin (CWBio, Beijing, China). After incubation by primary antibodies or their corresponding secondary antibodies, blots were developed using SuperSignal West Dura Persistence Substrate (Thermo Scientific).

Rong J., Jie Y, & Zhao H. (2022). m6A ‘writer’ KIAA1429 regulates the proliferation and migration of endothelial cells in atherosclerosis. Molecular Biotechnology.

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