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Phospho ire1α

Manufactured by Abcam
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Phospho-IRE1α is a lab equipment product that detects the phosphorylated form of the IRE1α protein. IRE1α is a key component of the unfolded protein response pathway. This product can be used to measure the activation of the IRE1α signaling pathway.

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Lab products found in correlation

γ h2ax, by Abcam (1 mentions) Ire1α, by Abcam (1 mentions) Xbp1s, by Abcam (1 mentions) Anti mouse or anti rabbit igg hrp linked secondary antibodies, by Cell Signaling Technology (1 mentions) Target lumiglo, by Cell Signaling Technology (1 mentions) Mre11, by Cell Signaling Technology (1 mentions) Bip grp78, by Santa Cruz Biotechnology (1 mentions) Tunicamycin, by Merck Group (1 mentions) Thapsigargin, by Merck Group (1 mentions) Caspase 3 p17, by Santa Cruz Biotechnology (1 mentions) Parp1 bml sa248, by Enzo Life Sciences (1 mentions)

Close spelling variants of this product

IRE1α (22 citations) Anti-phospho-IRE1α (5 citations) Phospho-IRE1α (ab48187) (3 citations) Phospho-IRE1α antibody (2 citations) Anti-phospho-IRE1α (Ser724) (2 citations) Anti–phospho-S724 IRE1α (2 citations)

3 protocols using phospho ire1α

1

Quantification of Cellular Stress Markers

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Total cellular protein was prepared with Pierce Lysis Buffer (Rockford, IL, USA) and analyzed using the Bradford method. The method applied is described in detail in our previous study.19 Primary antibodies, including antibodies against phospho‐ATM, cleaved caspase3, Rad50, Nbs1, Mre11, cleaved poly(ADP‐ribose) polymerase (PARP), IL‐6 (all 1:1000; Cell Signaling Technology, Danvers, MA, USA), IRE1α, phospho‐IRE1α, XBP‐1s and γH2AX (all 1:1000; Abcam), were incubated at 4°C overnight. Anti–mouse or anti–rabbit IgG HRP‐linked secondary antibodies (1:1000 dilution; Cell Signaling Technology), were added and incubated. Target proteins were developed with Target LumiGLO (Cell Signaling Technology) and photographed using a DNR BioImaging System (DNR, Israel).
Lyu X., Zhang M., Li G., Cai Y., Li G, & Qiao Q. (2019). Interleukin‐6 production mediated by the IRE1‐XBP1 pathway confers radioresistance in human papillomavirus‐negative oropharyngeal carcinoma. Cancer Science, 110(8), 2471-2484.
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2

Cellular ER Stress Pathway Inhibitors

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Cisplatin (cis-diammineplatinum(II) dichloride (Cat. No. P4394), 6-diazo-5-oxo-L-norleucine (DON, D2141), azaserine (A4142), D-(+)-glucosamine hydrochloride (G1515) and 2-deoxy-D-glucose (D8375) were purchased from Sigma (St. Louis, MO) and dissolved in distilled water. Tunicamycin (T7765) and thapsigargin (T9033) were also Sigma products and prepared with dimethyl sulfoxide (DMSO). IRE1 RNase inhibitor A106 (531399) was from Millipore (Burlington, MA) and dissolved in DMSO. The commercial antibodies for the following proteins were used : GFAT1 (ab125069), GFAT2 (ab176206), RL2 (ab2739, recognizing O-GlcNAc), phospho-IRE1α (serine 724, ab48187) from Abcam (Cambridge, MA); caspase 3 p17 (sc-271028, recognizing both uncleaved and cleaved forms), BiP/GRP78(glucose-regulated protein 78; sc-1050) from Santa Cruz Biotechnology (Santa Cruz, CA); CHOP (2895), Phospho-eIF2α (serine 51, 9721), BiP/GRP78 (3177) from Cell Signaling; β-actin (A2103) and β-tubulin (T6074) from Sigma; PARP1 (BML-SA248) from Enzo Life Sciences (Farmingdale, NY) and XBP1s (647502) from Biolegend (San Diego, CA).
Chen W., Chinh Do K., Saxton B., Leng S., Filipczak P., Tessema M., Belinsky S.A, & Lin Y. (2019). Inhibition of the hexosamine biosynthesis pathway potentiates cisplatin cytotoxicity by decreasing BiP expression in non-small cell lung cancer cells. Molecular carcinogenesis, 58(6), 1046-1055.
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3

Western Blot Analysis of Apoptosis Markers

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Cellular proteins were extracted from tumour tissues and lung cancer cells using RIPA lysis buffer. The proteins was separated on a 5-15% SDS-PAGE gel,then transferred to PVDF membranes, blocked with 5% non-fat milk and incubated with the indicated primary antibodies, including antibodies against cleaved caspase-3, PERK, ATF6, eIF2α, phospho-eIF2α, CHOP, phospho-JNK and β-actin (CST, USA) and antibodies against phospho-PERK and ATF4 (Affinity Biosciences) and phospho-IRE1α (Abcam, USA), overnight at 4 °C. Subsequently, the membranes were incubated with secondary antibodies at room temperature for 1.5 h. The immune signals were visualized with an ECL kit.
Tang Z., Du W., Xu F., Sun X., Chen W., Cui J., Tang W., Yang F., Teng F., Lin J., Liu B, & Dong J. (2022). Icariside II enhances cisplatin-induced apoptosis by promoting endoplasmic reticulum stress signalling in non-small cell lung cancer cells. International Journal of Biological Sciences, 18(5), 2060-2074.
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