Antibodies used for immunoblotting were: beta-actin (C4, Chemicon), ATM (MAT3; gift from Y. Shiloh, Tel-Aviv, Israel), pKAP1 S824 (Bethyl laboratories), pCHK2 T68 (Cell Signalling), pATM S1981 (Epitomics), Brca1 (OP92 and OP93, Calbiochem, La Jolla, CA), Nbs1 (NB100-143, Novus, Littleton, CO), Rap80 (70822, Abcam, Cambridge, MA), pBrca1 S1387 (A300-007A, Bethyl, Montgomery, TX), pBrca1 S1423 (2838, Abcam, Abcam, Cambridge, MA).
Pkap1 s824
Manufactured by Fortis Life Sciences
Sourced in Israel
The PKAP1 S824 is a laboratory equipment designed for protein kinase assays. It measures the activity of protein kinases, which are enzymes that play a crucial role in cellular signaling pathways. The PKAP1 S824 provides a platform for researchers to study the regulation and function of protein kinases in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Patm s1981, by Abcam (2 mentions)
Pchk2 t68, by Cell Signaling Technology (1 mentions)
Rap80, by Abcam (1 mentions)
Nbs1 nb100 143, by Novus Biologicals (1 mentions)
β actin, by Merck Group (1 mentions)
Dna pkcs, by Cell Signaling Technology (1 mentions)
Rad50, by Cell Signaling Technology (1 mentions)
Cenpf, by Abcam (1 mentions)
Pdna pkcs s2056, by Cell Signaling Technology (1 mentions)
Vinculin, by Cell Signaling Technology (1 mentions)
Mre11, by Cell Signaling Technology (1 mentions)
β catenin, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
4 protocols using pkap1 s824
1
Immunoblotting with DNA Damage Antibodies
2
Protein Extraction and Western Blotting
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For preparation of protein extracts, cells were lysed in cell extraction buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 2 mM EGTA, 2 mM EDTA, 25 mM NaF, 25 mN beta-glycerophosphate, 0.1 mM Na3VO4, 0.1 mM PMSF, 5 µg/ml leupeptin, 1 µg/ml aprotinin), 0.2% Triton X-100, 0.3% NonidetP-40 for 45 min on ice. Protein extracts were cleared through centrifugation at 19500xg (13200 rpm) for 15 min, and supernatants were separated through SDS-PAGE and transferred to nitrocellulose membranes by wet blot technique. Antibodies to TPT1 (Abcam, #ab133568, 1:50000), p-KAP1 S824 (Bethyl, #A300-767A-1, 1:2000–1:5000), p-CHEK2 S19 (Cell Signalling, #2666, 1:500–1:1000), p21 (Cell Signalling, #2846, 1:1000), ATM (Epitomics, #1549–1, 1:1000), DNA-PK (Calbiochem, #NA57T, 1:600) and β-actin (Sigma, #A5441, 1:3000) were applied over night at 4 °C after blocking with 5% skim milk in PBST for 1 h. Anti-mouse and anti-rabbit horseradish peroxidase-labelled secondary antibodies were purchased from GE Healthcare. Visualization of immunoreactive bands was performed with ECL and X-ray films.
3
Antibody-based Analysis of DNA Damage Repair
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Antibodies against the following proteins were used for immunofluorescence (IF) and Western blotting (WB): γH2AX (05–636 JBW301; Upstate Biotechnology), CENPF (ab5; Abcam), RPA (NA-18, Ab-2; Calbiochem for IF, LS-C38952; LifeSpan BioSciences for WB), pRPA S4/8 (A300-245A; Bethyl Laboratories), pDNA-PKcs S2056 (ab18192), DNA-PKcs (12311; Cell Signaling), Ku80 (2180 (C48E7); Cell Signaling), pKAP-1 S824 (A300-767A; Bethyl Laboratories), KAP-1 (ab3831; Abcam), MRE11 (12D7, GTX70212; GeneTex), NBS1 (PC269, Ab-1; Oncogene), RAD50 (3427; Cell Signaling), pATM S1981 (2152–1; Epitomics), ATM (GTX70103 2C1; GeneTex), CtIP (A300-488A; Bethyl Laboratories), and β-tubulin (ab21058; Abcam). Ku70, XLF, XRCC4, and Ligase IV antibodies were kind gifts from Dr. Penny Jeggo. Immunoblotting was performed as described previously [17 (link)].
4
Protein Extraction and Western Blotting
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Whole cell and tissue extracts were prepared in RIPA buffer (150 mM NaCl; 1% NP-40; 0.5% C24H39O4Na; 0.1% SDS; 50 mM Tris-HCl, pH 8.0), protein concentration was determined by BCA assay (Pierce), extracts were resolved by SDS-PAGE and transferred using standard procedures. Primary antibodies were: MRE11 (Cell Signaling, #4895); γH2AX and H2AX (EMD Millipore, #05–636 and #07–627); pKAP1S824 (Bethyl, #A300–767A); Vinculin (Cell Signaling, #4650); TOPOI (BD Biosciences, #556597); Cyclin D1 (Santa Cruz, #sc-8396); β-catenin (Santa Cruz, #sc-7199); and GAPDH (Cell Signaling, #2118). Secondary antibodies for western blots were IRDye-conjugated goat anti-rabbit or anti-mouse (Li-Cor Biosciences).
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