Analysis of DNA Damage Response Proteins

Nuclear and total cell lysates were obtained as described before^{11 (link)} and analyzed by SDS-PAGE using primary antibodies against: ATM (Santa Cruz Biotechnology #sc-135663), CtIP (Abcam #ab-70163), 53BP1 (Abcam #ab-175933), SLFN11 (Santa Cruz Biotechnology #sc-515071), BRCA1 (ThermoFisher Scientific #MA1–23164), BRCA2 (Santa Cruz Biotechnology #sc-28235), PALB2 (Proteintech #14340–1-AP), RAD51 (Abcam #ab-88572), RAD52 (Santa Cruz Biotechnology #sc-365341), RAD54 (Santa Cruz Biotechnology #sc-374598), DNA-PKcs (Bethyl #A300–518A), Ku70 (Santa Cruz Biotechnology #sc-17789), Ku80 (ThermoFisher Scientific #MA5–15873), DNA ligase 4 (ThermoFisher Scientific #PA5–40826), PARP1 (Santa Cruz Biotechnology #sc-74470), PARP2 (Santa Cruz Biotechnology #sc-393310), PARP3 (Santa Cruz Biotechnology #sc-390771), DNA ligase 3 (Santa Cruz Biotechnology #sc-135883), Polθ (MyBioSource #MBS9612322), lamin B (Abcam #ab-16048–100), and β-actin (Santa Cruz Biotechnology #sc-47778) and the following secondary antibodies conjugated to HRP (horseradish peroxidase): goat anti-rabbit (EMD Millipore #12–348) and goat anti-mouse (EMD Millipore #AP181P). ZNF251 western analysis was performed with ZNF251 antibody (Proteintech cat# 25601–1-AP) and GAPDH antibody (Cell signaling technology cat#2118). For quantification of western analysis, ImageJ software was used to measure the density of the protein bands.

Free full text: Click here

Li H., Chatla S., Liu X., Vekariya U., Kim D., Walt M., Lian Z., Morton G., Feng Z., Yang D., Liu H., Reed K., Childers W., Yu X., Madzo J., Chitrala K.N., Skorski T, & Huang J. (2023). Haploinsufficiency of ZNF251 causes DNA-PKcs-dependent resistance to PARP inhibitors in BRCA1-mutated cancer cells. Research Square.

Publication Preprint 2023

SLFN11 BRCA1 BRCA2 RAD51 RAD52 RAD54 DNA-PKcs PARP1 DNA ligase 3 β-actin AP181P GAPDH antibody

53bp1 Actin Antibodies Antibody Brca1 Brca2 Cat 1 Cell Ctip Dna ligase 3 Dna ligase 4 Dna pkcs Gapdh Goat Horseradish peroxidase Ku70 Lamin b Mouse Palb2 Parp1 Parp2 Parp3 Protein Rabbit Rad52 Sds page

Corresponding Organization :

Other organizations : Coriell Institute For Medical Research, Temple College, Temple University, Inovio Pharmaceuticals (United States), Shanghai Jiao Tong University, University of Houston

Top 5 similar protocols

Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of: ATM, CtIP, 53BP1, SLFN11, BRCA1, BRCA2, PALB2, RAD51, RAD52, RAD54, DNA-PKcs, Ku70, Ku80, DNA ligase 4, PARP1, PARP2, PARP3, DNA ligase 3, Polθ, lamin B, β-actin, ZNF251, GAPDH

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None 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!