Rabbit anti ph2ax

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-pH2AX is a laboratory equipment product that detects phosphorylation of the histone H2AX at serine 139, which is a marker of DNA double-strand breaks.

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Lab products found in correlation

Aperio system, by Leica (1 mentions) Rabbit anti nitrotyrosine, by Merck Group (1 mentions) Rabbit anti psmad3, by Abcam (1 mentions) Cellsens dimension software, by Olympus (1 mentions) Goat anti il 6, by R&D Systems (1 mentions) Rabbit anti hemagglutinin ha, by Cell Signaling Technology (1 mentions) Rabbit anti tnf α, by Novus Biologicals (1 mentions) Rabbit anti ki67, by Cell Signaling Technology (1 mentions) Zen lite, by Zeiss (1 mentions) Mouse anti parp1, by Merck Group (1 mentions)

Spelling variants of this product

Anti-pH2AX (21 citations) Anti-pH2AX Ser139 rabbit mAb (2 citations)

3 protocols using rabbit anti ph2ax

Immunohistochemical Analysis of Tumor Samples

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Formalin-fixed, paraffin-embedded tumor samples were sectioned at a thickness of 3 μm, dewaxed, hydrated and stained with hematoxylin and eosin (H&E) or processed for immunohistochemistry with rabbit anti-human c-Myc (Abcam), rabbit anti-pH2AX (Cell Signaling Technology, MA, USA) or rabbit anti-nitrotyrosine (Millipore) antibodies.
Positive signal was revealed by 3,3’-diaminibenzidine (Roche) stainings. Sections were finally counterstained with Carazzi’s hematoxylin before analysis by light microscopy. Images were acquired with the automatic high-resolution scanner Aperio System (Leica Biosystems, Wetzlar, Germany, EU) and image analysis was carried out using the open-source ImageJ software.

Giacomini A., Taranto S., Rezzola S., Matarazzo S., Grillo E., Bugatti M., Scotuzzi A., Guerra J., Di Trani M., Presta M, & Ronca R. (2020). Inhibition of the FGF/FGFR System Induces Apoptosis in Lung Cancer Cells via c-Myc Downregulation and Oxidative Stress. International Journal of Molecular Sciences, 21(24), 9376.

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Comprehensive Immunohistochemistry Staining Protocol

Cited 1 time

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We performed immunohistochemistry (IHC) staining as previously described.^{15 (link)} Primary antibodies used were rabbit anti-Ki67 (1:500) (Cell Signaling Technology, 12202), rabbit anti-NFκB-p105/p50 [p Ser337] (1:300) (Novus Biologicals, NB100–82074), rabbit anti-pSmad3 (1:400, Abcam, ab52903), rabbit anti-pH2AX (1:100) (Cell Signaling Technology, 9718), rabbit anti-myeloperoxidase (Ready-to-Use, Dako, GA51161–2), rabbit anti-hemagglutinin (HA) (1:700) (Cell Signaling Technology, 3724), rabbit anti-IL-1β (1:500) (Novus Biologicals, NB600–633), rabbit anti-TNF-α (1:200) (Novus Biologicals, NBP1–19532), mouse anti-CCL2 (1:50) (Novus Biologicals, MAB28171), and goat anti-IL-6 (1:40) (R&D, AF1609) antibodies. Nuclear pSmad3-, NFκB p50-, or pH2AX-positive cells were quantified as the percent of positive cells per total epithelial cell count (excluding sloughed epithelial cells induced by irradiation). Nuclear Ki67 was quantified as positive cells per length of basement membrane. For pH2AX staining, cells with more than 3 nuclear foci were defined as pH2AX-positive cells. Myeloperoxidase-positive cells were quantified per square millimeter epithelial and stroma area above the muscle layer, and sequential 10 × images along the basement membrane were quantified and averaged per sample using Olympus cellSens Dimension software.

Boss M.K., Ke Y., Bian L., Harrison L.G., Lee B.I., Prebble A., Martin T., Trageser E., Hall S., Wang D.D., Wang S., Chow L., Holwerda B., Raben D., Regan D., Karam S.D., Dow S., Young C.D, & Wang X.J. (2021). Therapeutic Intervention Using a Smad7-Based Tat Protein to Treat Radiation-Induced Oral Mucositis. International journal of radiation oncology, biology, physics, 112(3), 759-770.

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Immunofluorescence Microscopy of DNA Damage

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1 × 10⁵ cells were seeded on glass cover slips prior to treatment in a 24-well plate. After treatment, cells were fixed with 4% paraformaldehyde for 20 min. They were subsequently washed with PBS and blocked with 5% FBS/PBS with 0.3% Triton-X. The cells were incubated overnight at 4 °C with primary antibody (Mouse Anti-PARP1, 1:1000 (Sigma); Rabbit Anti-pH2AX, 1:500 (Cell signalling)) dissolved in blocking solution. After washing with PBS, the cells were incubated for 1 h with secondary antibodies (Goat Anti-Mouse DyLight 488, 1:2000 and Goat Anti-Rabbit Alexa Fluor 568, 1:1000) at room temperature. After the PBS wash steps, the cells were mounted with Roti Mount^® Vectashield containing DAPI and analysed using fluorescence microscopy (Zen Lite, Carl Zeiss, Jena, Germany). Quantification was performed using CellProfiler 3.0 software.

Sankaranarayanan R.A., Peil J., Vogg A.T., Bolm C., Terhorst S., Classen A., Bauwens M., Maurer J., Mottaghy F, & Morgenroth A. (2022). Auger Emitter Conjugated PARP Inhibitor for Therapy in Triple Negative Breast Cancers: A Comparative In-Vitro Study. Cancers, 14(1), 230.

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