The following antibodies were used in this study: PRP19 (ab27692, 1:400 IF, 1:1000 WB), ATM pS1981 (ab81292, 1:1000 WB), DNAPKcs pS2056 (ab18192, 1:1000 WB) and RPA32 pT21 (ab109394, 1:5000 WB) antibodies were from Abcam, CHK1 (SC-8404, 1:1000 WB), Myc (SC-40, 1:1000 WB) and HA (SC-7392, 1:1000 WB) antibodies were from Santa Cruz; RPA32 antibodies (MA1–26418, 1:500 IF, 1:1000 WB) were from Thermo, ATM (A300–299A, 1:1000 WB), RPA32 pS4/8 (A300–245A, 1:1000 WB), RPA32 pS33 (A300–246A, 1:5000 WB), RFWD3 (A301–397A, 1:250 IF, 1:1000 WB) antibodies were from Bethyl; ubiquitin antibodies were from Covance (MMS-257P, 1:1000 WB); CDC5L antibodies were from BD-Bioscience (612362, 1:1000 WB); anti-FLAG antibodies were from Sigma (F1804, 1:800 IF, 1:1000 WB); CHK1 pS345 (2348, 1:1000 WB), GAPDH (5174, 1:1000 WB), DNAPKcs (12311, 1:1000 WB) and tubulin (2144, 1:1000 WB) antibodies were from Cell Signaling.
Rpa32 ps4 8
Manufactured by Fortis Life Sciences
The RPA32 pS4/8 is a lab equipment product designed for scientific research. It serves as a tool for the analysis and study of proteins. The core function of this product is to facilitate the detection and quantification of specific phosphorylation states of the RPA32 protein subunit. This information can contribute to a better understanding of cellular processes and signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Chk1 ps345, by Cell Signaling Technology (2 mentions)
Rpa32 pt21, by Abcam (2 mentions)
Chk2 pt68, by Cell Signaling Technology (2 mentions)
Anti flag antibody, by Merck Group (1 mentions)
Dnapkcs ps2056, by Abcam (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Dna pkcs, by Cell Signaling Technology (1 mentions)
Myc sc 40, by Santa Cruz Biotechnology (1 mentions)
Rpa32 ps33, by Fortis Life Sciences (1 mentions)
Rfwd3, by Fortis Life Sciences (1 mentions)
α tubulin, by Merck Group (1 mentions)
Thymidine, by Merck Group (1 mentions)
Nocodazole, by Merck Group (1 mentions)
Mk 1775, by Selleck Chemicals (1 mentions)
Sb218078, by Bio-Techne (1 mentions)
Rpa70, by Abcam (1 mentions)
Roscovitine, by Selleck Chemicals (1 mentions)
Rpa32, by Fortis Life Sciences (1 mentions)
Ve 821, by Selleck Chemicals (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
4 protocols using rpa32 ps4 8
1
DNA Damage Response Antibodies
2
Investigating Cell Cycle Regulation
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The following chemical inhibitors were used at the indicated concentrations unless stated otherwise: WEE1 inhibitor MK-1775 (500 nM; Selleck Chemicals), CHK1 inhibitor SB 218078 (2 μM; Tocris), ATR inhibitor VE-821 (20 μM; Selleck Chemicals), CDK1 inhibitor RO-3306 (10 μM; Calbiochem), CDK inhibitor Roscovitine (20 μM; Selleck Chemicals), thymidine (2 mmol/L; Sigma-Aldrich), and nocodazole (100 ng/mL; Sigma-Aldrich), IdU (10 μg/ml, Sigma-Aldrich), CIdU (10 μg/ml, Sigma-Aldrich). The following antibodies were used: CHK1 (Santa Cruz Biotechnology), CHK1-pS345 (Cell Signaling), CHK2 (Santa Cruz Biotechnology), CHK2-pT68 (Cell Signaling), phospho-Histone-H2AX-Ser139 (γH2AX; Millpore), CDK1 (Santa Cruz Biotechnology), CDK1-pTyr15 (Cell Signaling), RPA70 (Abcam), phospho-(Ser) CDKs Substrate (pCDK Substrate; Cell Signaling), CDK2 (Santa Cruz Biotechnology), CDK2-pTyr15 (Abcam), RPA32-pT21 (Abcam), RPA32-pS33 (Abcam), RPA32-pS4/8 (Bethyl Laboratories), RPA32 (Bethyl Laboratories), WEE1 (Santa Cruz Biotechnology), α-Tubulin (Sigma-Aldrich), CIdU (rat anti-BrdU; Accurate Chemical), IdU (mouse anti-BrdU; Becton Dickinson).
3
Antibody Profiling for Cell Cycle Regulation
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Commercially available antibodies obtained from Cell Signaling against the indicated proteins were: Cdc2/CDK1 (Cat # 9112S), CDK1-P-Y15 (Cat # 4539S), Histone H3-P-S28 (Cat # 9713S), p53-P-S15 (Cat # 9284), SMC1-P-957 (Cat # 4805S), SMC1 (Cat # 4802S), CDC25A (Cat # 3652S) and Chk2-P-T68 (Cat # 2661S). Antibodies obtained from Santa Cruz Biotechnology were CDC25A F-6 (Cat # sc -7389), CDC25C H-150 (Cat # sc-5620), Cdc2 (Cat # sc-54). Others were against actin (Cat. 109 # MA515739, Pierce), RPA32-P-S4/8 (Cat # A300-245A, Bethyl), cyclin A (Cat # 06-138, Upstate) γH2AX (Cat # 05-636, Millipore), Chk2 Clone 7 (Cat # 05-649, Millipore), cyclin B1 (Cat. # 05-373, Millipore), and CDK1 (Cat. #Ab18, Abcam). Secondary antibodies have previously been described. Additional antibodies used for immunofluorescence include: cyclin B1 (Cat # 4138S, Cell signaling), Alexa 488 (anti-mouse Cat # A11029, anti-rabbit Cat # A11034, Invitrogen) and Alexa 568 (anti-mouse Cat # A11031, anti-rabbit Cat # A11036, Invitrogen). NS1 CE10 and NS1 91W were described previously [7] (link).
4
Isolation and Maintenance of Mouse Embryonic Fibroblasts
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MEF cells were isolated from embryonic day 13.5 (E13.5) embryos as described previously (Castillo et al. 2014 (link)). Primary MEFs were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin antibiotics and maintained in 3% O2/5% CO2 in an incubator at 37°C. Immortalized MEFs were generated by spontaneous immortalization following the 3T3 protocol as described previously (Castillo et al. 2014 (link)). PD20 and PD20/FANCD2 cells were maintained in RPMI1640 medium with 15% FBS. U2OS cells were grown in McCoy's 5A medium with 10% FBS. CAPAN-1 cells were cultured in Iscove's modified Dulbecco's medium (IMDM) (Hyclone, Thermo Scientific) with 20% FBS. Antibodies used in this study included Abro1 (Hu et al. 2011 (link)), BRCC36 (Abcam), BRE (Hu et al. 2011 (link)), NBA1 (Hu et al. 2011 (link)), 53BP1 (Novus Biologicals), BLM (Bethyl Laboratories), BrdU (BD Bioscience), RAD51 (Calbiochem), RPA32pS4/8 (Bethyl Laboratories), p-CHK1 (Cell Signaling), CHK1 (Santa Cruz Biotechnolgoy), γH2AX (Ser139; Upstate Biotechnology), Flag (Sigma), BRCA2 (Abcam, ab27976), γ-tubulin (Sigma), DNA2 (Proteintech, 18727-1-AP), FANCD2 (Santa Cruz Biotechnology, sc-20022), and H3 (Abcam). Chemicals used in this study included BrdU (Sigma), APH (Sigma), HU (Sigma), MMC (Sigma), and camptothecin (CPT) (Sigma).
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