Immunoblot Analysis of RNase L, Histones, and Cytoskeletal Proteins

Immunoblot analysis was performed as described in [13 (link)]. Mouse anti-RNase L antibody 2E9 (Novus Biologicals catalog no. NB100-351) was used at 1:1500. Histone H3 antibody (Thermo Fisher Scientific; NB500-171) was used at 1:1000. Mouse anti-alpha tubulin (Abcam: ab18251) was used at 1:1000. Rabbit anti-GAPDH (Cell Signaling Technology: 2118L) was used at 1:2000. Anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)–linked antibody (Cell Signaling Technology: 7074S) was used at 1:3000. Anti-mouse IgG, HRP-linked antibody (Cell Signaling Technology: 7076S) was used at 1:10,000 Crude nuclear and cytoplasmic fractionation was performed as described in [58 (link)].

Free full text: Click here

Burke J.M., Ripin N., Ferretti M.B., St Clair L.A., Worden-Sapper E.R., Salgado F., Sawyer S.L., Perera R., Lynch K.W, & Parker R. (2022). RNase L activation in the cytoplasm induces aberrant processing of mRNAs in the nucleus. PLOS Pathogens, 18(11), e1010930.

Publication 2022

Histone H3 antibody Mouse anti-alpha tubulin Rabbit anti-GAPDH Anti-rabbit immunoglobulin G (IgG), horseradish peroxidase (HRP)–linked antibody

Alpha tubulin Anti immunoglobulin Antibody Biologicals Cytoplasmic Fractionation Gapdh Histone h3 Horseradish peroxidase Immunoblot analysis Immunoglobulin g Mouse Novus Rabbit Rnase l

Corresponding Organization : Colorado State University

Top 5 similar protocols

Variable analysis

independent variables

Crude nuclear and cytoplasmic fractionation

dependent variables

RNase L protein levels
Histone H3 protein levels
Alpha tubulin protein levels
GAPDH protein levels

control variables

Immunoblot analysis protocol described in [13]
Antibody concentrations for RNase L (1:1500), Histone H3 (1:1000), Alpha tubulin (1:1000), GAPDH (1:2000)
Anti-rabbit IgG (1:3000) and anti-mouse IgG (1:10,000) HRP-linked antibodies

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!