Anti gh2ax

Manufactured by Cell Signaling Technology

Anti-gH2AX is a primary antibody product offered by Cell Signaling Technology. It is used to detect the phosphorylation of histone H2AX, which is an important marker of DNA double-strand breaks.

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Lab products found in correlation

Anti chk1, by Cell Signaling Technology (2 mentions) Anti cdk1, by Santa Cruz Biotechnology (2 mentions) Anti mcm2, by Santa Cruz Biotechnology (2 mentions) P chk1, by Cell Signaling Technology (2 mentions) Anti pcdk1, by Cell Signaling Technology (2 mentions) Anti parp1, by Cell Signaling Technology (2 mentions) Anti pol 2, by Merck Group (1 mentions) Mab374, by Merck Group (1 mentions) Ab109315, by Abcam (1 mentions) Ab124707, by Abcam (1 mentions) Anti lamin b1, by Abcam (1 mentions) Anti fancd2, by Santa Cruz Biotechnology (1 mentions) Anti cyclin b1, by Santa Cruz Biotechnology (1 mentions) Ab16048, by Abcam (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti phospho erk, by Santa Cruz Biotechnology (1 mentions) Anti tubulin, by Cell Signaling Technology (1 mentions) Anti lamin a c, by Abcam (1 mentions) Anti actin, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

GH2AX (5 citations)

6 protocols using anti gh2ax

Comprehensive Immunoblotting Methodology

Cited 1 time

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Immunoblotting was performed as described previously (35 (link)). The following antibodies were used at a concentration of 0.1 to 0.5 μg/ml: anti-pMCM2 (3378-1; Epitomics Inc.), anti-MCM2 (sc-9839; Santa Cruz Biotechnology), anti–pPOL II (04-1571; Millipore), anti–POL II (05-952; Millipore), anti-Lamin B1 (ab16048; Abcam), anti-FANCD2 (sc-20022; Santa Cruz Biotechnology), anti-pCDC6 (ab76422; Abcam), anti-CDC6 (ab109315; Abcam), pChk1 (#2348; Cell Signaling Technology), anti-Chk1 (#2345; Cell Signaling Technology), anti-pCDK1 (#9111; Cell Signaling Technology), anti-CDK1 (sc-954; Santa Cruz Biotechnology), anti-gH2AX (#2577; Cell Signaling Technology), anti-PARP1 (#9542; Cell Signaling Technology), anti–cyclin B1 (sc-752; Santa Cruz Biotechnology), anti-CDC7 (sc-56274; Santa Cruz Biotechnology), anti-DBF4 (ab124707; Abcam), and anti-GAPDH (MAB374; Chemicon). Immunoblotted proteins were visualized by chemiluminescence.

Iwai K., Nambu T., Dairiki R., Ohori M., Yu J., Burke K., Gotou M., Yamamoto Y., Ebara S., Shibata S., Hibino R., Nishizawa S., Miyazaki T., Homma M., Oguro Y., Imada T., Cho N., Uchiyama N., Kogame A., Takeuchi T., Kurasawa O., Yamanaka K., Niu H, & Ohashi A. (2019). Molecular mechanism and potential target indication of TAK-931, a novel CDC7-selective inhibitor. Science Advances, 5(5), eaav3660.

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Western Blotting of Cell Extracts

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For Western blotting, cells were washed in PBS, scraped in SDS gel-loading buffer, and boiled for 5 min. Proteins of total extracts were separated by SDS-PAGE and transferred to nitrocellulose membranes. Monoclonal antibody against HA was purchased from BioLegend. Anti-HA goat was from Bethyl Laboratories. Anti-emerin and anti-lamin A/C antibodies were from Abcam and Cell Signaling. Anti-tubulin, anti-gH2AX, and anti-ERK were from Cell Signaling. Anti-GAPDH, anti-actin, anti-lamin B, anti-RanGAP, anti-RanBP, and anti-phospho-ERK antibodies were from Santa Cruz Biotechnology. Anti-VP24 was from Biorbyt. Anti-VP35 and anti-NP were from Genetex. Anti-BAF antibody was from Abcam.

Vidal S., Sánchez-Aparicio M., Seoane R., El Motiam A., Nelson E.V., Bouzaher Y.H., Baz-Martínez M., García-Dorival I., Gonzalo S., Vázquez E., Vidal A., Muñoz-Fontela C., García-Sastre A, & Rivas C. (2021). Expression of the Ebola Virus VP24 Protein Compromises the Integrity of the Nuclear Envelope and Induces a Laminopathy-Like Cellular Phenotype. mBio, 12(4), e00972-21.

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Immunoblotting of Cell Signaling Proteins

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Immunoblotting was performed as described previously (44 (link)). The following antibodies were used at 0.1 to 0.5 μg/ml: anti-pMCM2 (3378-1; Epitomics Inc.), anti-MCM2 (sc-9839; Santa Cruz Biotechnology), pChk1 (#2348; Cell Signaling Technology), anti-Chk1 (#2345; Cell Signaling Technology), anti-pCDK1 (#9111; Cell Signaling Technology), anti-CDK1 (sc-954; Santa Cruz Biotechnology), anti-pBRCA1 (#9009; Cell Signaling Technology), anti-pp53 (#9286; Cell Signaling Technology), anti-pATM (2152-1; Epitomics Inc.), anti-p21 (#2947; Cell Signaling Technology), anti-gH2AX (#2577; Cell Signaling Technology), anti-p53 (sc-126; Santa Cruz Biotechnology), anti-PARP1 (#9542; Cell Signaling Technology), anti-BRCA1 (sc-6954; Santa Cruz Biotechnology), anti-CDC7 (sc-56274; Santa Cruz Biotechnology), anti-DBF4 (ab124707; Abcam), and anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (MAB374; Chemicon). Immunoblotted proteins were visualized using chemiluminescence.

Iwai K., Nambu T., Kashima Y., Yu J., Eng K., Miyamoto K., Kakoi K., Gotou M., Takeuchi T., Kogame A., Sappal J., Murai S., Haeno H., Kageyama S.I., Kurasawa O., Niu H., Kannan K, & Ohashi A. (2021). A CDC7 inhibitor sensitizes DNA-damaging chemotherapies by suppressing homologous recombination repair to delay DNA damage recovery. Science Advances, 7(21), eabf0197.

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Comprehensive Western Blot and Immunofluorescence Analysis of DNA Damage Response Proteins

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For western blot of endogenous proteins, anti-gH2AX (Cell Signaling Technology (CST), MA #9718, 1:1000), anti-Actin B (CST, MA #4970, 1:1000), anti-p-ATM S1981 (CST, MA #13050, 1:1000), anti-total ATM (CST, MA #2873, 1:1000), anti-total H2AX (CST, MA #2595, 1:1000), anti-RING1A (CST, MA #13069, 1:1000), anti-RING1B (Santa Cruz (SC), CA sc-101109, 1:1000), anti-H2AK119ub1 (CST, MA #8240,1:1000), anti-XPC ( SC, CA sc-74410, 1:1000), anti-lamin B (SC, CA sc-6216, 1:1000 ; SC, CA sc-374015, 1:1000), anti-XPA (SC, CA sc-28353, 1:1000), anti-pRPA32 S33 (Bethyl Laboratories, TX A300-246A-M, 1:1000), anti-RPA32 (CST, MA #2208,1:1000), antiphospho-Chk1 S345 (CST, MA #2348, 1:1000) and anti-total Chk1 (CST, MA #2360, 1:1000) antibodies were used. For immunofluorescence, anti-gH2AX (CST,MA #9718, 1:100), anti-RING1A (Abcam, CA ab175149, 1:100), anti-RPA32 (CST, MA #2208, 1:100), anti-Rad51 (Novus biological, CO NB100-148, 1:100), anti-H2AK119ub1 (Millipore Sigma, MA 05-678,1:100) and secondary Alexa Conjugate (CST, MA rat #4416, 1:1000, mouse #8890, 1:500, rabbit #4412, 1:1000 and, rabbit #8889,1:500) antibodies were used.

Sriramkumar S., Matthews T.D., Ghobashi A.H., Miller S.A., VanderVere-Carozza P.S., Pawelczak K.S., Nephew K.P., Turchi J.J., & O’Hagan H.M. (2020). Platinum-Induced Ubiquitination of Phosphorylated H2AX by RING1A is Mediated by Replication Protein A in Ovarian Cancer.

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DNA Damage Assessment by γ-H2AX Foci

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The cells were seeded on coverslips in 24-well plates and treated with cisplatin. After incubation with a drug-free medium for the indicated time, cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. Then, the cells were blocked with 3% bovine serum albumin and immunostained overnight with antig-H2AX (Cell Signaling Technology, 9718). Subsequently, the cells were incubated with a Cy3-conjugated secondary antibody and counterstained with DAPI (Roche). Images were captured using an A1-Si Nikon confocal laser scanning microscope. Only those cells with five or more foci were considered g-H2AX foci positive.

Xiao M., Guo J., Xie L., Yang C., Gong L., Wang Z, & Cai J. (2020). Let-7e Suppresses DNA Damage Repair and Sensitizes Ovarian Cancer to Cisplatin through Targeting PARP1. Molecular cancer research : MCR, 18(3).

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Antibody detection of apoptosis markers

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Antibodies against IGFBP3 (sc-9028) and b-actin (sc-47778) were purchased from Santa Cruz Biotechnology. Anti-caspase-3, anti-poly (ADP-ribose) polymerase (anti-PARP), and anti-gH2AX antibodies were purchased from Cell Signaling Technology.

Chen C.H., Chen P.Y., Lin Y.Y., Feng L.Y., Chen S.H., Chen C.Y., Huang Y.C., Huang C.Y., Jung S.M., Chen L.Y, & Wei K.C. (2020). Suppression of tumor growth via IGFBP3 depletion as a potential treatment in glioma. Journal of neurosurgery, 132(1).

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