DNA Damage Response Protein Analysis

Western blot was performed as described previously (19 (link), 20 (link)), using pATM (Ser1981) (D6H9), MRE11, RAD50, NBS1, PARP-1, pAKT pCHK2 (Thr68) (C13C1), γH2AX (Cell Signaling, Danvers, MA), and β-Actin (8H10D10) antibodies (Cell Signaling). Appropriate horseradish peroxide-conjugated secondary antibodies (Cell Signaling) were used, and proteins were detected with the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare Bio-Sciences, Pittsburgh, PA). The blotted membranes were stripped and re-probed with ATM (D2E2), AKT, and CHK2 (D9C6) antibodies (Cell Signaling). Protein bands were captured and quantified by Chemi Doc^TM MP Imaging System (Bio-Rad).

Free full text: Click here

Zhao J., Nguyen L.N., Nguyen L.N., Dang X., Cao D., Khanal S., Schank M., Thakuri B.K., Ogbu S.C., Morrison Z.D., Wu X.Y., Li Z., Zou Y., El Gazzar M., Ning S., Wang L., Moorman J.P, & Yao Z.Q. (2019). ATM Deficiency Accelerates DNA Damage, Telomere Erosion, and Premature T Cell Aging in HIV-Infected Individuals on Antiretroviral Therapy. Frontiers in Immunology, 10, 2531.

Publication 2019

RAD50 PARP-1 β-Actin (8H10D10) Appropriate horseradish peroxide-conjugated secondary antibodies Amersham ECL Prime Western Blotting Detection Reagent CHK2 (D9C6) Chemi DocTM MP Imaging System

Actin Antibodies Horseradish Membranes Parp 1 Peroxide Protein Rad50

Corresponding Organization : James H. Quillen VA Medical Center

Top 5 similar protocols

Variable analysis

independent variables

Western blot protocol (as described previously in references 19 and 20)

dependent variables

PATM (Ser1981)
MRE11
RAD50
PARP-1
PCHK2 (Thr68)
γH2AX
β-Actin

control variables

Horseradish peroxide-conjugated secondary antibodies
Amersham ECL Prime Western Blotting Detection Reagent
Chemi Doc^TM MP Imaging System

controls

Positive controls: ATM (D2E2), AKT, and CHK2 (D9C6) antibodies

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!