Rabbit anti histone h2ax

Manufactured by Cell Signaling Technology

Sourced in United States

The Rabbit anti-Histone H2AX is a lab equipment product. It is an antibody that recognizes the Histone H2AX protein, which is a variant of the H2A histone family and plays a role in the DNA damage response pathway.

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2 protocols using rabbit anti histone h2ax

Western Blot Analysis of Cell Signaling

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Whole cell lysates for Western Blot were prepared with the SDS-sample buffer. Proteins were separated by SDS-PAGE and transferred onto the PVDF membrane (Millipore/Fisher Scientific, Pittsburg, PA), Darmstadt, Germany). Antibodies for immunoblotting from Cell Signaling Technology (Dancers, MA) would include rabbit anti-Bmal1 (#14020), rabbit anti-CLOCK (#5157), rabbit anti-phospho-p53 (Ser15) [#9284], rabbit anti-p21 Waf1/Cip1 (#2947), rabbit anti-Histone H2AX (#2595), rabbit anti-cleaved PARP (#9541), and rabbit anti-phospho-CHK1 (Ser 345) (#2348). Other antibodies include mouse anti-p53 (sc-126; Santa Cruz Biotechnology Inc., Santa Cruz, CA), rabbit anti-Keratin 10 (Poly19054; Biolegend, San Diego, CA), mouse anti-phospho-Histone H2AX (Ser139) (05–636; Millipore/Fisher Scientific), and mouse anti-α-tubulin (T9026; Sigma-Aldrich, St. Louis, MO). HRP-conjugated secondary antibodies were from Santa Cruz Biotechnology Inc. Chemiluminescence images were acquired using an Amersham Imager 600 from GE Healthcare Life Sciences (Pittsburgh, PA) or iBright FL1000 (Invitrogen-Life Technology/Fisher Scientific, Pittsburgh, PA). The level of target proteins was quantified by densitometry scanning with the ImageJ software and normalized to the amount of α-tubulin.

Sun Y., Wang P., Li H, & Dai J. (2018). BMAL1 and CLOCK proteins in regulating UVB-induced apoptosis and DNA damage responses in human keratinocytes. Journal of cellular physiology, 233(12), 9563-9574.

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Western Blotting Analysis of GPR30, Iba1, and TLR4

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Expression of the GPR30, Iba1, and TLR4 proteins in penumbra and primary microglia was measured by Western blotting as previously described [2 (link)]. The extracted proteins were separated by 10% SDS-PAGE and electrically transferred to polyvinylidene difluoride membranes. Then, the membranes were blocked with TBST containing 5% nonfat dry milk for 1 h at room temperature. The following primary antibodies were used: rabbit anti-GPR30 (1:1000; Abcam, England), rabbit anti-Iba1 (1:500; Wako, Japan), rabbit anti-TLR4 (1:200; Santa Cruz, USA), rabbit anti-NF-κB (Cell Signaling Technology; 1:500), mouse anti-β-actin (1:1000; Cell Signaling Technology, USA), rabbit anti-GAPDH (1:1000; Cell Signaling Technology, USA), and rabbit anti-Histone H2A.X (Cell Signaling Technology; 1:1000). The membranes were shaken at 60 rpm/min at 4 °C overnight and incubated with an IRDye secondary anti-rabbit or mouse antibody (Thermo Scientific, USA) for 1 h. Protein bands were visualized using the LI-COR Odyssey System (LI-COR Biotechnology, USA).

Zhang Z., Qin P., Deng Y., Ma Z., Guo H., Guo H., Hou Y., Wang S., Zou W., Sun Y., Ma Y, & Hou W. (2018). The novel estrogenic receptor GPR30 alleviates ischemic injury by inhibiting TLR4-mediated microglial inflammation. Journal of Neuroinflammation, 15, 206.

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