Immunoblotting was performed as described19 (link) with the following antibodies: 53BP1 (175933, Abcam; 1:1000; 100-304, Novus Biological; 1:1000); BRCA1 (MAB22101, R+D systems; 1:500); BRCA2 (OP95, Millipore; 1:500); CHK1 (8408, Santa Cruz; 1:1000); pCHK1 (2341, Cell Signaling Technology; 1:1000); CHK2 (611570, BD; 1:1000); Flag-tag (M2, Sigma; 1:1000); γ-Tubulin (GTU488, Sigma; 1:20,000); GFP (11814460001, Sigma; 1:1000); HA (3724, Cell Signaling Technology; 1:20,000); HSP70 (610608, BD; 1:1000); MAD2L2/REV7 (180579, Abcam; 612266, BD; 1:1000); Myc-tag (9B11, Cell Signaling Technology; 1:1000); OBFC1/STN1 (89250, Abcam; 1:1000; SC-376450, Santa Cruz; 1:1000); PRIM1 (10773-1-AP, Proteintech; 1:1000); RIF1 (#1240, de Lange Lab; 1:1000); SHLD1 (PA5-59280, Thermo-Fisher; 1:1000); SNAP tag (9310, NEB; 1:1000); TRF2 (#1254, de Lange Lab; 1:5,000). Affinity-purified peptide antibodies against mouse SHLD1 and SHLD2 proteins (Chapman Lab, unpublished; 1:1000) were generated by Eurogentec.
Protocol detail
Immunoblotting Antibodies for DNA Repair
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
53bp1
Antibodies
Antibodies affinity
Biological
Brca1
Brca2
Hsp70
Mouse
Novus
Peptide
Proteins
Tubulin
Corresponding Organization :
Other organizations : Rockefeller University, MRC Weatherall Institute of Molecular Medicine, Medical Research Council, University of Oxford
Top 5 similar protocols
Variable analysis
independent variables
- Antibodies used for immunoblotting: 53BP1, BRCA1, BRCA2, CHK1, pCHK1, CHK2, Flag-tag, γ-Tubulin, GFP, HA, HSP70, MAD2L2/REV7, Myc-tag, OBFC1/STN1, PRIM1, RIF1, SHLD1, SNAP tag, TRF2, mouse SHLD1 and SHLD2
- Protein expression levels as measured by immunoblotting
- Immunoblotting protocol described in the referenced publication (#19)
- No specific positive or negative controls were mentioned in the input information.
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.
Sign up for free or login to display all annotations
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!