All chemicals and solvents used in the present study were of highest grade available. InuA with a purity higher than 95% was prepared as reported previously [34 (link)]. The antibodies against human MDM2 (Ab-2), p21 (Ab-1), and p-H2AX (Ser139) were bought from EMD Chemicals (Gibbstown, NJ, USA). The antibodies against human p53 (DO-1), Cdk2 (M2), Cdk4 (H-22), Cdk6 (C-21), cyclin D1 (DCS-6), cyclin E (HE12), c-Myc (0.N.222), Bax (N-20), Bcl-2 (100), PARP (H-250), Chk1 (G-4), Chk2 (B-4), and ATR (N-19) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The antibodies against human p-Chk1 (Ser317), p-Chk2 (Thr68), and p-p53 (Ser15) were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-human NFAT1 (1/NFAT-1) antibody was sourced from BD Biosciences (San Jose, CA, USA) and the anti-human β-actin (AC-15) antibody was purchased from Sigma (St. Louis, MO, USA). The goat anti-mouse IgG (H+L) and goat anti-rabbit IgG (H+L) antibodies were bought from Bio-Rad (Hercules, CA, USA). The NFAT1 and MDM2 siRNA pool and control siRNA pool were obtained from Thermo Scientific (Rockford, IL, USA).
Atr n 19
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The ATR (N-19) is a laboratory equipment product offered by Santa Cruz Biotechnology. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach. Further information about the intended use or interpretation of the product's capabilities is not available.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg h l, by Bio-Rad (2 mentions)
Goat anti rabbit igg h l, by Bio-Rad (2 mentions)
Anti human nfat1 1 nfat 1 antibody, by BD (2 mentions)
Anti human β actin ac 15 antibody, by Merck Group (2 mentions)
Biomax mr film, by Kodak (2 mentions)
Xar film, by Kodak (2 mentions)
γ tubulin gtu 88, by Merck Group (2 mentions)
P p53 ser15, by Cell Signaling Technology (1 mentions)
Mops buffer, by Thermo Fisher Scientific (1 mentions)
Jbw301, by Merck Group (1 mentions)
Btf 608a, by Fortis Life Sciences (1 mentions)
Cyclin b1 gns1, by Santa Cruz Biotechnology (1 mentions)
Atm 2c1, by Santa Cruz Biotechnology (1 mentions)
Patr s428, by Cell Signaling Technology (1 mentions)
Brca2 h 300, by Santa Cruz Biotechnology (1 mentions)
Ab8580, by Abcam (1 mentions)
Ab1791, by Abcam (1 mentions)
Ab9045, by Abcam (1 mentions)
Ab8898, by Abcam (1 mentions)
Ab1220, by Abcam (1 mentions)
8 protocols using atr n 19
1
Molecular Mechanisms of Cellular Regulation
2
Whole Cell Extract Immunoblotting
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Whole cell extracts were prepared by lysing cells in two volumes of Lysis Buffer (250 mM NaCl, 5 mM ethylenediaminetetraacetic acid (EDTA), 50 mM HEPES, 0.1% v/v NP40). A total of 60 μg of whole cell extract (WCE) was resolved on 4–12% NOVEX gels using MOPS buffer (Invitrogen). Protein was transferred to PVDF membrane (Invitrogen) and blotted for BCLAF1 (BTF 608A, Bethyl Labs); THRAP3 (A300–956A, Bethyl Labs), GAPDH (ab9485, Abcam), pATM (S1981) (ab79891 Abcam), ATM (2C1, Santa Cruz), pATR (S428) (2853, Cell Signaling), ATR (N-19, Santa Cruz), 53BP1 (NB100–34, Novus Biologicals), γH2AX (S139) (JBW301, Millipore), cyclin B1 (GNS1) (sc-245, Santa Cruz), pP53 (S15) (9284S, Cell Signaling), pKAP1 (S824) (A300–767A, Bethyl Labs), FANCD2 (FI17) (sc200–22, Santa Cruz), BRCA2 (H-300) (sc-8326, Santa Cruz), FANCL (B-11) (sc-137067, Santa Cruz), PALB2 (H-45) (sc-382436, Santa Cruz) and Rad51 (3C10) (sc-53428, Santa Cruz).
3
Histone Modifications and DNA Damage Signaling
4
Immunoblotting for Telomere Regulatory Factors
5
Reagents and Antibodies for Protein Analysis
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LinA was prepared as described in our previous study[48 ]. All chemicals and solvents used in the present study were of highest grade available. Fetal bovine serum (FBS) was obtained from Atlanta Biologicals (Lawrenceville, GA, USA). The penicillin/streptomycin preparation was bought from Corning (Manassas, VA, USA). The anti-human NFAT1 (1/NFAT-1) antibody was purchased from BD Biosciences (San Jose, CA, USA). The antibodies against human MDM2 (Ab-2), p21 (Ab-1), and p-H2AX (Ser139) were sourced from EMD Chemicals (Gibbstown, NJ, USA). The antibodies against human p53 (DO-1), Cdk2 (M2), Cdk4 (H-22), Cdk6 (C-21), cyclin D1 (DCS-6), cyclin E (HE12), c-Myc (0.N.222), Bax (N-20), Bcl-2 (100), PARP (H-250), Chk1 (G-4), Chk2 (B-4), and ATR (N-19) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The antibodies against human p-Chk1 (Ser317), p-Chk2 (Thr68), and p-p53 (Ser15 (link)) were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-human β-actin (AC-15) antibody was bought from Sigma (St. Louis, MO, USA) and the goat anti-mouse IgG (H+L) and goat anti-rabbit IgG (H+L) antibodies were obtained from Bio-Rad (Hercules, CA, USA).
6
Quantitative Protein Analysis by SDS-PAGE and Western Blot
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Total protein was extracted from exponentially growing cells at passage 8–10 and 40 g/mL were resolved by SDS-PAGE using a 4–15% gradient gel (Bio-Rad Laboratories). After transfer and blocking overnight at 4 °C in Odyssey Blocking Buffer (Li-Cor, Lincoln, NE, USA) proteins were detected by primary antibodies against BRCA1 [2A-9] (1:500, kindly provided by Stephen Smith, Leibnitz Institute, Jena, Germany), ATR [N-19] (Santa Cruz, St. Cruz, CA, USA, 1:1000), CHK1 [2G1D5] (Cell Signaling, Danvers, MA, USA, 1:750), RAD51 [14B4] (1:2.000, GeneTex, Irvine, CA, USA), MRE11A [12D7] (Abcam, Cambridge, UK, 1:500), pCHK1 [Ser296] (Cell Signaling, 1:1000), -actin [AC-74] (1:50.000, Sigma, St. Louis, MO, USA). Primary antibodies were detected with IRDYE 680 conjugated anti-mouse IgG, IRDYE 800 conjugated anti-rabbit IgG (Li-Cor, 1:7500), IRDYE 680 conjugated anti-rabbit IgG (Licor, 1:7.500 or 15.000) or IRDYE 800 conjugated anti-mouse IgG (Li-Cor 1:7.500 or 15.000). Quantitative and qualitative analysis was done by using Li-Cor Odyssey (Li-Cor, Lincoln, NE, USA).
7
DNA Damage Response Protein Assay
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ATM (Cell Signaling 2873s), ATM S1981 (Cell Signaling 4526L), ATR (N19) (Santa Cruz sc-1807), BRCA2 [15 (link)], DNA-PKcs S2056 (Abcam ab18192), gamma-H2AX [16 (link)], 53BP1 [17 (link)], RAD51 [15 (link)], Actin (Sigma A5441), GAPDH (Chemicon MAB374), H3 (Cell Signaling 9715s), CHK1 (Santa Cruz sc-8048), and mouse anti-BrdU (BD Biosciences 347-580 B44) were used at 1:500 dilution. All secondary antibodies were purchased from Jackson Immuno Research Laboratories and were used at 1:3000 dilution.
8
Immunoblotting Protein Detection Protocol
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