αpstat1

Manufactured by Cell Signaling Technology

The αpSTAT1 is a laboratory equipment product that detects the phosphorylated form of STAT1 protein. It is used in research applications to study cellular signaling pathways.

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Lab products found in correlation

α stat1, by Cell Signaling Technology (2 mentions) α actin, by Merck Group (1 mentions) α gapdh, by Santa Cruz Biotechnology (1 mentions) α h3k27me3, by Cell Signaling Technology (1 mentions) Eed226, by Selleck Chemicals (1 mentions) Gsk503, by Selleck Chemicals (1 mentions) α p38 mapk, by Cell Signaling Technology (1 mentions)

2 protocols using αpstat1

Immunoblotting analysis of PRC2 inhibitors

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Antibodies used for immunoblotting analysis in this study include: αEZH2 (BD Biosciences, #612667), αActin (Sigma-Aldrich, #A5441), αH3K27me3 (Cell Signaling, #9733), αH3 (Cell Signaling, #9715), αpSTAT2 (Cell Signaling, #88410), αSTAT2 (Santa Cruz, #sc-1668 and Cell Signaling, #72604), αpSTAT1 (Cell Signaling, #9167), αSTAT1 (Cell Signaling, #9172) and αGAPDH (Santa Cruz, #sc-365062). Antibodies that were applied in other assays, such as ChIP, fluorescent immunohistochemistry and flow cytometry, are specified in the corresponding sections.
In this study, the following inhibitors of the PRC2 activity were included: GSK503 (Selleckchem, #S7804), GSK126 (Xcessbio, #60071), EPZ-6438 (Xcessbio, #60122) and EED226 (Selleckchem, #S8496). The interferon (IFN) ligands used in this study are: IFNβ, IFNγ and IL-29, and their details are provided in the section “Neutralization of IFN ligands” below.

Hong J., Lee J.H., Zhang Z., Wu Y., Yang M., Liao Y., de la Rosa R., Scheirer J., Pechacek D., Zhang N., Xu Z., Curiel T., Tan X., Huang T.H, & Xu K. (2022). PRC2-mediated epigenetic suppression of type I IFN-STAT2 signaling impairs antitumor immunity in luminal breast cancer. Cancer research, 82(24), 4624-4640.

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Characterizing Immune Signaling Pathways

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Gp96, calreticulin, and HSP90 were isolated from murine livers as previously described^{10 (link),22 (link)}. The quality and purity of the gp96 preparations was vigilantly verified by SDS-PAGE gel and immunoblotting and is consistent with our previous studies.
For immunoblots, cells were lysed with NP-40 and lysates were run on SDS-PAGE gels and transferred to nitrocellulose membranes. Membranes were probed with the following antibodies

α

-STAT1(14944),

α

-pSTAT1 S727(9177),

α

-pSTAT1 Y701(9167),

α

-pp38 MAPK(9167),

α

-p38 MAPK(9216),

α

β

-actin(8457), (Cell Signaling) and

α

-SHP2(sc-7384) (Santa Cruz). Immunoblots comparing different cell types and timepoints were run with all on a singular membrane when possible. Membranes were developed using a Proteinsimple imager and quantitated using Alpha View software.
Luminex (eBioscience) consisted of the single analytes: IL-12, IL-18, IL-15/IL-15R, IFN-

α

, IP-10, and RANTES. Samples were analyzed using the manufacturer’s protocol and a Bioplex II Luminex machine.
CXCL10 and CCL5 ELISAs were purchased from eBioscience and samples were analyzed following the manufacturer’s protocol.

Sedlacek A.L., Kinner-Bibeau L.B., Wang Y., Mizes A.P, & Binder R.J. (2021). Tunable heat shock protein-mediated NK cell responses are orchestrated by STAT1 in Antigen Presenting Cells. Scientific Reports, 11, 16106.

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