Acetylation analysis of TMV proteins

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).

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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.

Publication 2022

Immobilon-P anti-rabbit secondary antibody beta-actin

Acetylation Anti antibody Antibody Beta actin Electrophoresis Immobilon p Membrane Mouse Protein Rabbit Sds page Sodium dodecyl sulfate Western blot

Corresponding Organization : Chinese Academy of Agricultural Sciences

Other organizations : China Tobacco

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Variable analysis

independent variables

Time points of inoculation with PBS and TMV solutions

dependent variables

Acetylation of proteins detected by Western blot analysis

control variables

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) for protein isolation
Transfer of proteins to a polyvinylidene fluoride (PVDF) membrane
Use of TMV coat protein (cp) antibody, anti-rabbit secondary antibody, and beta-actin antibody for analysis
Use of anti-acetyl-lysine primary antibody and anti-mouse secondary antibody conjugated to HRP for detection of acetylation
Analysis of Western blot results using ImageJ (v.1.52a)

positive controls

Proteins isolated from samples inoculated with PBS

negative controls

Not explicitly mentioned

Annotations

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