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α γh2ax

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The α-γH2AX is a laboratory equipment product used in the detection and quantification of DNA double-strand breaks. It is a key marker for the cellular response to DNA damage. The product allows for the visualization and analysis of the phosphorylation of the histone variant H2AX, which is one of the earliest cellular responses to the presence of DNA double-strand breaks.

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Lab products found in correlation

α h2a, by Abcam (3 mentions) α macroh2a1, by Merck Group (2 mentions) α h2ax, by Abcam (2 mentions) α h2b, by Abcam (2 mentions) α brca1, by Santa Cruz Biotechnology (2 mentions) α gapdh, by Santa Cruz Biotechnology (2 mentions) α phospho s139 h2ax, by Merck Group (2 mentions) α phospho s139 h2ax s139, by Merck Group (2 mentions) α atrx, by Cell Signaling Technology (2 mentions) α brdu, by BD (2 mentions) Img 124a, by Merck Group (2 mentions) Alexa fluor 488 or 647, by Thermo Fisher Scientific (2 mentions) Goat anti rabbit igg h l hrp, by Thermo Fisher Scientific (2 mentions) Goat anti mouse igg h l hrp, by Thermo Fisher Scientific (2 mentions) Normal mouse igg, by Merck Group (2 mentions) Img 124a, by Novus Biologicals (2 mentions) α pan akt, by Cell Signaling Technology (1 mentions) α phospho akt ser473, by Cell Signaling Technology (1 mentions) Goat α mouse irdye 680rd, by LI COR (1 mentions) α tubulin, by Merck Group (1 mentions)

Spelling variants of this product

γH2AX (349 citations) Mouse α-γH2AX (2 citations)

6 protocols using α γh2ax

1

Antibody Profiling for DNA Damage Response

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Primary antibodies used were: α-ubiquityl-PCNA (D5C7P; Cat# 13439), α-PCNA (PC-10; Cat# 2586), α-pan-Akt (Cat# 4691), α-phospho-Akt (Ser473; Cat# 9271), α-phospho-GSK3B (Ser9; Cat# 9336), α-phospho-PRAS40 (Thr246; Cat# 2997), α-RAD18 (Cat# 9040) and α-SMC-1 (Cat# 4802) from Cell Signaling Technology; α-BRCA1 (Ab-1) from Oncogene Research; α-PCNA (PC-10, Cat# sc-56) from SCBT; α-γH2AX (Cat# 05-636-1) from Millipore; α-CPD (Cat# NMDND001) from Cosmo Bio; α-Tubulin (Cat# T9026) from Sigma-Aldrich. Secondary antibodies used were: α-mouse Alexa Fluor 594 from Jackson ImmunoResearch; goat α-mouse IRDye 680RD (Cat# P/N 925-68070) and goat α-rabbit IRDye 800CW (Cat# P/N 925-32211) from LI-COR Biosciences. Nuclei were stained with DAPI (Cat# D9542) from Sigma-Aldrich.
Villafañez F., García I.A., Carbajosa S., Pansa M.F., Mansilla S., Llorens M.C., Angiolini V., Guantay L., Jacobs H., Madauss K.P., Gloger I., Gottifredi V., Bocco J.L, & Soria G. (2019). AKT inhibition impairs PCNA ubiquitylation and triggers synthetic lethality in homologous recombination-deficient cells submitted to replication stress. Oncogene, 38(22), 4310-4324.
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2

Chromatin Fractionation and Western Blot Analysis

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Cells were lysed and harvested with Laemmli buffer, followed by 8 min of incubation at 99°C. The chromatin fraction was obtained after a 5-min extraction with ice-cold CSK buffer [10 mM Pipes (pH 7.5), 100 mM NaCl, 300 mM sucrose, 1 mM EGTA, 3 mM MgCl2, and 2% Triton X-100]. The following antibodies were used: α-Chk1 at 1:1000 (Santa Cruz Biotechnology, sc-8408), α-γH2AX at 1:4000 (Millipore, 05-636), α-phospho-KAP1 Ser824 at 1:4000 (Bethyl Laboratories, A300-767A), α-Mus81 at 1:1000 (Santa Cruz Biotechnology, sc-53382), α-CDC45 at 1:1000 (Santa Cruz Biotechnology, sc-20685), α-KAP1 at 1:4000 (Bethyl Laboratories, A300-274A), α-phospho-RPA Ser4/8 at 1:8000 (Bethyl Laboratories, A300-245A), α-H2B (histone 2B) at 1:2000 (Santa Cruz Biotechnology, sc-515808), and α-actin at 1:20,000 (Sigma-Aldrich, A2066). Incubations with secondary antibodies (Jackson ImmunoResearch) and enhanced chemiluminescence (ECL) detection (GE Healthcare) were performed according to the manufacturers’ instructions. Western blot images were acquired with ImageQuant LAS4000 (GE Healthcare), which allows the capture and the quantification of images within a linear range.
Calzetta N.L., Besteiro M.A, & Gottifredi V. (2020). Mus81-Eme1–dependent aberrant processing of DNA replication intermediates in mitosis impairs genome integrity. Science Advances, 6(50), eabc8257.
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3

ChIP-Seq and Western Blot Antibodies

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The following antibodies were used for ChIP: α-macroH2A1.2 (Millipore MABE61), α-phospho-S139-H2AX (Millipore 05–636), α-H2B (Abcam ab52484), α-H2A (Abcam ab18255), TRF2 (Novus Biologicals IMG-124A) and normal mouse IgG (Millipore 12–371). Primary antibodies for western blotting were: α-macroH2A1.2 (Millipore MABE61), α-phospho-S139-H2AX (S139) (Millipore 05–636), α-BRCA1 (Santa Cruz sc-6954), α-ATRX (Cell signaling 14820s), α-GAPDH (Santa Cruz sc-32233), α-H2AX (Abcam ab20669). Secondary antibodies were goat anti-mouse IgG (H+L)-HRP (Invitrogen 31430) and goat anti-rabbit IgG (H+L)-HRP (Invitrogen 31460). Primary antibodies for IF were α-BRCA1 (Santa Cruz sc-6954), α-γ-H2AX (Millipore 05–636, Abcam ab11174), α-BrdU (BD Biosciences 555627), and TRF2 (Novus Biologicals IMG-124A), secondary antibodies were goat-anti-mouse or goat-anti-rabbit IgG (H+L) coupled to Alexa Fluor 488 or 647 (Life Technologies).
Kim J., Sun C., Tran A.D., Chin P.J., Ruiz P.D., Wang K., Gibbons R.J., Gamble M.J., Liu Y, & Oberdoerffer P. (2019). The macroH2A1.2 histone variant links ATRX loss to alternative telomere lengthening.. Nature structural & molecular biology, 26(3), 213-219.
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4

Chromatin Profiling using CUT&RUN and Immunoblotting

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Antibodies used for CUT&RUN are αH2A (Abcam, 15653), αFLAG (Sigma, F1804), αmouse rb IgG (Jackson ImmunoResearch, 315-005-003). Primary antibodies used for immunoblotting include: αγH2Ax (Milipore, ab26350), αFLAG (Sigma, F7425), αH2B (CST, D2H6), αH4 (CST, D2X4V), αH3 (Immunoway, YM3038), αNAP1L1 (Abcam, ab33076), αNAP1L2 (Abnova, H00004674-D01), αSPT16 (Santa Cruz, sc-377028), αSSRP1 (Santa Cruz, sc-74536), αHSC70 (Santa Cruz, sc-7298), αC23 (Santa Cruz, sc-55486), αH2BK120ub (CST, D11), αH2AK119ub (CST, D11), αH3K4me3 (Active Motif, 61379), αH3K36me3 (Immunoway), αH3K36me2 (CST, 29015), αH3K9me3 (Abcam, ab8898), αH3K27me3 (CST, 9733), αH4K16ac (Immunoway, YK0014), αH4K20me3 (Active Motif, 39671), αH3K27ac (Active Motif, 39685), αH3K79me3 (Immunoway, YM3091), αH3K9ac (Immunoway, YK0006), αHA (Santa Cruz, SC7392), αβ-Actin (Immunoway, YM3028). αFLAG M2 Affinity Gel (Sigma), αH2A (Abcam, 15653), and αRpb1 (CST, 14958) were used for ChIP.
Kang T.Z., Zhu L., Yang D., Ding D., Zhu X., Wan Y.C., Liu J., Ramakrishnan S., Chan L.L., Chan S.Y., Wang X., Gan H., Han J., Ishibashi T., Li Q, & Chan K.M. (2021). The elevated transcription of ADAM19 by the oncohistone H2BE76K contributes to oncogenic properties in breast cancer. The Journal of Biological Chemistry, 296, 100374.
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5

Antibodies Used in Cellular Assays

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The following antibodies were used: α-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Novus Biologicals; no. NB300-285IR), α-PARP1 (Santa Cruz Biotechnology; no. sc-25780), α-APE1 (Novus Biologicals; no. NB100-116, Cell Signaling; no. 4128), α-PNKP (78 (link)), α-DNA Polβ (Proteintech; no. 18003–1-AP, Santa Cruz Biotechnology; no. sc-376581), α-XRCC1 (Neomarkers; no. MS 434-P, Abcam; no. ab134056, Invitrogen; no. MA5-13412), α-LSD1 (Abcam; no. ab17721, Millipore; no. 05–939), α-TET1 (Millipore; no. 09–872, Thermo Fisher Scientific; no. MA5-16312), α-MLH1 (Cell Signaling; no. 3515), α-PMS2 (BD Biosciences; no. 556415), α-XRCC4 (Santa Cruz Biotechnology; no. sc-271087), α-γH2AX (Millipore; no. 05–636-I), and α-8-oxoG (Trevigen; no. 4354-MC-050).
Sengupta S., Wang H., Yang C., Szczesny B., Hegde M.L, & Mitra S. (2020). Ligand-induced gene activation is associated with oxidative genome damage whose repair is required for transcription. Proceedings of the National Academy of Sciences of the United States of America, 117(36), 22183-22192.
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6

ChIP-Seq and Western Blot Antibodies

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The following antibodies were used for ChIP: α-macroH2A1.2 (Millipore MABE61), α-phospho-S139-H2AX (Millipore 05–636), α-H2B (Abcam ab52484), α-H2A (Abcam ab18255), TRF2 (Novus Biologicals IMG-124A) and normal mouse IgG (Millipore 12–371). Primary antibodies for western blotting were: α-macroH2A1.2 (Millipore MABE61), α-phospho-S139-H2AX (S139) (Millipore 05–636), α-BRCA1 (Santa Cruz sc-6954), α-ATRX (Cell signaling 14820s), α-GAPDH (Santa Cruz sc-32233), α-H2AX (Abcam ab20669). Secondary antibodies were goat anti-mouse IgG (H+L)-HRP (Invitrogen 31430) and goat anti-rabbit IgG (H+L)-HRP (Invitrogen 31460). Primary antibodies for IF were α-BRCA1 (Santa Cruz sc-6954), α-γ-H2AX (Millipore 05–636, Abcam ab11174), α-BrdU (BD Biosciences 555627), and TRF2 (Novus Biologicals IMG-124A), secondary antibodies were goat-anti-mouse or goat-anti-rabbit IgG (H+L) coupled to Alexa Fluor 488 or 647 (Life Technologies).
Kim J., Sun C., Tran A.D., Chin P.J., Ruiz P.D., Wang K., Gibbons R.J., Gamble M.J., Liu Y, & Oberdoerffer P. (2019). The macroH2A1.2 histone variant links ATRX loss to alternative telomere lengthening.. Nature structural & molecular biology, 26(3), 213-219.
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