Anti γh2ax

Manufactured by Novus Biologicals

Sourced in United States

Anti-γH2AX is a primary antibody used for the detection of phosphorylated histone H2AX (γH2AX), a marker of DNA double-strand breaks. It is a useful tool for the study of DNA damage response pathways.

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Lab products found in correlation

Hoechst 33342, by Merck Group (2 mentions) Anti rrna, by Novus Biologicals (2 mentions) Anti rad51, by Abcam (2 mentions) Anti flag, by Merck Group (2 mentions) Anti puromycin, by Merck Group (2 mentions) Anti chk2, by Cell Signaling Technology (1 mentions) Slc7a11, by Cell Signaling Technology (1 mentions) Trieasy total rna extraction reagent, by Yeasen (1 mentions) Antibodies against cox 2, by Cell Signaling Technology (1 mentions) Fer 1, by Merck Group (1 mentions) Anti gpx4, by Abcam (1 mentions) Mitotracker red cmxros, by Yeasen (1 mentions) 2 7 dichlorodihydrofluorescein diacetate h2dcfda, by Thermo Fisher Scientific (1 mentions) Bodipy 581 591 c11 c11 bodipy, by Thermo Fisher Scientific (1 mentions) Anti acsl4, by Abcam (1 mentions) Anti acrolein, by Abcam (1 mentions) Anti γh2ax, by Merck Group (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

Spelling variants of this product

γH2AX (15 citations) Mouse anti-γH2AX (2 citations)

6 protocols using anti γh2ax

Mitotic Spindle Protein Localization

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Tubulin-specific DM1α (Sigma-Aldrich), Anti-centromere (CREST,
Dartmouth), hec1/NDC80-specific (Novus Biologicals), anti-γ-H2AX- (Novus
Biologicals), GFP-specific (William Wickner), anti- pChk2 S19 and S33/35 (Cell
Signaling Technology), anti-Chk2 (Cell Signaling Technology), anti-pPlk1-Thr210
(Cell Signaling Technology), anti-P-Aurora-A antibodies were used at dilutions
of 1:1000. 1:10000 for anti-GFP-antibody).

Bakhoum S.F., Kabeche L., Murnane J.P., Zaki B.I, & Compton D.A. (2014). DNA damage response during mitosis induces whole chromosome mis-segregation. Cancer discovery, 4(11), 1281-1289.

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Ferroptosis Induction and Detection

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atRAL, Fer-1 (catalog no. SML0583), DFO (catalog no. D9533), 4',6-diamidino-2-phenylindole (DAPI) and Hoechst 33342 were purchased from Sigma-Aldrich (Saint Louis, MO, USA). 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) and BODIPY 581/591 C11 (C11-BODIPY) were obtained from ThermoFisher Scientific (Eugene, OR, USA). Mitotracker Red CMXRos and TRIeasy total RNA extraction reagent were obtained from Yeasen (Shanghai, China). Antibodies against COX2 (catalog no. 12282S), SLC7A11 (catalog no. 98051S), cleaved caspase-3 (catalog no. 9664S and 9661S), and GAPDH (catalog no. 5174S) were provided by Cell Signaling Technology (Danvers, MA, USA). Anti-GPX4 (catalog no. ab125066), anti-ACSL4 (catalog no. ab155282), and anti-acrolein (catalog no. ab48501) were purchased from Abcam (South Cambs, England, UK). Anti-γH2AX (catalog no. 05-636) was purchased from Millipore (Billerica, MA, USA). Anti-γH2AX (catalog no. NB100-384) was provided by Novus Biologicals (Littleton, CO, USA).

Chen C., Chen J., Wang Y., Liu Z, & Wu Y. (2020). Ferroptosis drives photoreceptor degeneration in mice with defects in all-trans-retinal clearance. The Journal of Biological Chemistry, 296, 100187.

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Immunofluorescence Staining for γH2AX and TDP-43

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After deparaffinisation, antigen retrieval in 10 mM citrate buffer with 0.1% Tween for 30 min at 80 °C was performed. Sections were blocked with normal donkey serum for 30 min and then stained with anti-γH2AX (1:200, Novus Biologicals) or anti-human TDP-43 antibodies (1:1000, Proteintech) for 24 h at 4 °C, and after three washes with PBS, the tissues were incubated with AlexaFluor 488/564 (1:200, Molecular Probes) conjugated secondary antibodies for 2 h at 4 °C. The nuclei were counterstained with Hoechst 33342 (Sigma-Aldrich).

Konopka A., Whelan D.R., Jamali M.S., Perri E., Shahheydari H., Toth R.P., Parakh S., Robinson T., Cheong A., Mehta P., Vidal M., Ragagnin A.M., Khizhnyak I., Jagaraj C.J., Galper J., Grima N., Deva A., Shadfar S., Nicholson G.A., Yang S., Cutts S.M., Horejsi Z., Bell T.D., Walker A.K., Blair I.P, & Atkin J.D. (2020). Impaired NHEJ repair in amyotrophic lateral sclerosis is associated with TDP-43 mutations. Molecular Neurodegeneration, 15, 51.

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Dissecting TGF-β Signaling Pathways

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The adenoviral expression vectors for IKKβ, SMAD7, β-GAL, GFP and GFP-Cre were from Drs. Yi Zheng at the Cincinnati Children’s Hospital, Yinling Hu at the National Cancer Institute, and Chia-yang Liu at Indiana University. The reporter plasmids, NF-κB-luc, SBE-Luc, AP-1-Luc, and the TGFβ1, Tgfβ2 and Tgfβ3 promoter-luc were obtained from Drs. Edward B. Leof at Mayo Clinic and Alvaro Puga at the University of Cincinnati (Tojima et al., 2000 (link)). Expression vector for SOD2 was from Dr. Shanglin Shi at the University of Kentucky and Bdm-c-Jun was described before (Geh et al., 2011 (link)). TGFβ1 was from PeproTech, NAC, BSO and DPI were from Sigma-Aldrich, and SB505124 was from EMD Millipore. The following antibodies were used in the study: anti- IKKα, -IKKβ, -IkBα, and -p-SMAD2 (Ser-465, 467) from Cell Signaling, anti-pan TGFβ from R&D Systems, anti-α-SMA from Abcam, anti-β-actin from Sigma-Aldrich, anti-γH2AX from Novus Biologicals, anti-PolII, -H3, -H3K27Me3, H3K9Me2, H3K9Ac and H3K4Me3 from EMD Millipore, and anti-p65, -c-Jun, and IgG from Santa Cruz Biotechnologies.

Chen L., Peng Z., Meng Q., Mongan M., Wang J., Sartor M., Chen J., Niu L., Medvedovic M., Kao W, & Xia Y. (2016). Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop. Protein & Cell, 7(5), 338-350.

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Antibody Immunoblotting Protocol

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Antibodies were purchased from respective companies: anti-FLAG (Sigma; rabbit, catalog no.: F7425; mouse, catalog no.: F1804), anti-γH2AX (Novus; rabbit, catalog no.: NB100-384; homemade, mouse), anti-RAD51AP1 (Proteintech; rabbit, catalog no.: 11255-1-AP), anti-rRNA (Novus; rabbit, catalog no.: NB100-662), IgG (CST; rabbit, catalog no.: 2729), antipuromycin (Sigma–Aldrich; mouse, catalog no.: 12D10), anti-GAPDH (CST; rabbit, catalog no.: 5174), anti-H3 (CST; rabbit, catalog no.: 4499S), anti-RAD51 (Abcam; rabbit, catalog no.: ab133534), and anti-RPA (Novus; rabbit, catalog no.: NBP1-23017).

Chen L., Gai X, & Yu X. (2024). Pre-rRNA facilitates the recruitment of RAD51AP1 to DNA double-strand breaks. The Journal of Biological Chemistry, 300(3), 107115.

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Comprehensive Antibody Characterization for Research

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Detailed information of antibodies used in this study: anti‐Flag (Sigma, Rabbit, F7425), anti‐TopBP1 (Santa Cruz, Mouse, sc‐271043; BETHYL, Rabbit, A300‐111A), anti‐γH2AX (Novus, Rabbit, NB100‐384; CST, Rabbit, 9718), anti‐SCP3 (Santa Cruz, Mouse, sc‐74569; Novus, Rabbit, NB300‐232), anti‐rRNA (Novus, Mouse, NB100‐662), anti‐RPL7A (ABclonal, Rabbit, A13713), anti‐RPS3 (ABclonal, Rabbit, A2533), anti‐FBL (CST, Rabbit, 2639S; Santa cruz, Mouse, sc‐166001), IgG (Rabbit, CST, 2729), IgG (Mouse, CST, 3420), anti‐ATR (BETHYL, Rabbit, A300‐137A), anti‐Chk1 (Novus, Rabbit, NB100‐464), anti‐pATR (T1989) (GeneTex, Rabbit, GTX128145), anti‐pChk1 (Ser345) (CST, Rabbit, 2348), anti‐GAPDH (CST, Mouse, 97 166), anti‐pH3 (Ser10) (CST, Rabbit, 9701), anti‐Puromycin (Millipore, Mouse, MABE343), anti‐GAPDH (HUABIO, Rabbit, ET1601‐4), anti‐Histone H3 (CST, Rabbit, 4499S), anti‐ATM (GeneTex, Mouse, GTX70107), anti‐pATM (S1981) (ROCKLAND, Mouse, 200‐301‐400), anti‐pRPA (Novus, Rabbit, NBP1‐23017), anti‐Rad9 (Santa Cruz, Mouse, sc‐74464), anti‐RAD51 (abcam, Rabbit, ab63801).

Xin D., Gai X., Ma Y., Li Z., Li Q, & Yu X. (2023). Pre‐rRNA Facilitates TopBP1‐Mediated DNA Double‐Strand Break Response. Advanced Science, 10(28), 2206931.

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