Muscle tissues and myotubes were homogenized in lysis buffer containing 20 mM HEPES (pH 7.2), 150 mM NaCl, 0.5% Triton X-100, 0.1 mM Na3VO4, 1 mM NaF, 1 mM 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF), and 5 mg/ml aprotinin (Sigma-Aldrich). The lysates were centrifuged at 15,000×g for 20 min at 4 °C, and the supernatants were subjected to SDS-PAGE followed by immunoblot analysis. Antibodies used are as follows: 4EBP1 (#9452), ATG5 (#12994), ATG7 (#8558), ATG12 (#4180), ATG16L (#8089), beclin1 (#3495), eIF2α (#9722), IRE1α (#3294), mTOR (#2983), p65 (#6956), PERK (#3192), S6K (#9202), SEK (#9152), SQSTM1 (#8025) phospho-4EBP1 (#9459), phospho-AKT S473 (#9271), phospho-GSK-3β (#9327), phosphor-mTOR (#5536), phospho-eIF2α (#9721), and phospho-PERK (#3179), phospho-JNK (#9251), phospho-SEK (#9151), phospho-S6K (#9206) from Cell Signaling Technologies; phospho-IRE1α (ab48187) from Abcam; and HA (sc-805), AKT (sc-1618), ATF6 (sc-22799), MYH (B-5, sc-376157), GSK-3β (sc-7291), and FABP3 (sc-58274) from Santa Cruz Biotechnology; and JNK (51-1570) from BD Bioscience; Laminin (L9393) from Sigma-Aldrich. The anti-GAPDH antibody was developed in our laboratory. Data were collected using Automatic X-ray Film Processor (JP-33, JPI America) or iBright FL1500 (Invitrogen).
Phospho ire1α ab48187
Manufactured by Abcam
Phospho-IRE1α (ab48187) is an antibody that detects the phosphorylated form of the IRE1α protein. IRE1α is a key component of the unfolded protein response, playing a crucial role in the cellular response to endoplasmic reticulum stress. This antibody can be used to measure the activation of the IRE1α pathway.
Automatically generated - may contain errors
Lab products found in correlation
P erk, by Cell Signaling Technology (1 mentions)
Laminin, by Merck Group (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Phospho eif2α, by Cell Signaling Technology (1 mentions)
Phospho gsk3β, by Cell Signaling Technology (1 mentions)
Phospho perk, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Phospho akt s473, by Cell Signaling Technology (1 mentions)
Phosphor mtor, by Cell Signaling Technology (1 mentions)
Phospho s6k, by Cell Signaling Technology (1 mentions)
Phospho sek, by Cell Signaling Technology (1 mentions)
Phospho 4ebp1, by Cell Signaling Technology (1 mentions)
Caspase 9 9502, by Cell Signaling Technology (1 mentions)
Chop 15204 1 ap, by Proteintech (1 mentions)
P jnk 4668, by Cell Signaling Technology (1 mentions)
Mitochondrial membrane potential assay kit with jc 1 c2006, by Beyotime (1 mentions)
3 protocols using phospho ire1α ab48187
1
Protein Expression Analysis in Muscle Tissues
2
Investigating Molecular Mechanisms of Endoplasmic Reticulum Stress
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Methylseleninic acid (MSeA,
purity >95%) and palmitic acid (PA) (P0500) were purchased from
Sigma-Aldrich
(St. Louis, MO). IRE1α inhibitor 4μ8C was purchased from
MCE (Shanghai, China). Primary antibodies specific for phospho-eIF2α
(3398), c-PARP (9546), Bip (3183), Bim (2933), and caspase-9 (9502)
were purchased from Cell Signaling Technology (Beverly, MA). Phospho-IRE1α
(ab48187) was purchased from Abcam (Cambridge, MA). CHOP (15204-1-AP)
was purchased from Protein Tech (Rosemont, IL). Secondary antibodies
specific for rabbit and mouse immunoglobulins were purchased from
MBL International Corporation (Woburn, MA).
purity >95%) and palmitic acid (PA) (P0500) were purchased from
Sigma-Aldrich
(St. Louis, MO). IRE1α inhibitor 4μ8C was purchased from
MCE (Shanghai, China). Primary antibodies specific for phospho-eIF2α
(3398), c-PARP (9546), Bip (3183), Bim (2933), and caspase-9 (9502)
were purchased from Cell Signaling Technology (Beverly, MA). Phospho-IRE1α
(ab48187) was purchased from Abcam (Cambridge, MA). CHOP (15204-1-AP)
was purchased from Protein Tech (Rosemont, IL). Secondary antibodies
specific for rabbit and mouse immunoglobulins were purchased from
MBL International Corporation (Woburn, MA).
3
Glycycoumarin-Induced ER Stress Signaling
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Glycycoumarin (GCM purity >99%) was purchased from BioBioPha. Sodium palmitate (P9767), cycloheximide (CHX) (C1988) and 3-MA (M9281) were purchased from Sigma-Aldrich. Primary-antibodies specific for Bip (3183), phospho-EIF2α (3597), phospho-PERK (3192), CHOP (2895), Bax (2772), Bak (12105), BCL2 (3869), PUMA (14570), Bim (2933), c-PARP (9548), caspase 9 (9502), c-caspase 3 (9929), c-caspase 9 (9929), phospho-GS (3891) and p-JNK (4668) were purchased from Cell Signaling Technology. Phospho-IRE1α (ab48187) was purchased from Abcam. Primary-antibody specific for β-actin (AT0001) was purchased from Action Biotech. Primary-antibodies specific for p62 (PM045), LC3 (PM036) and horseradish peroxidase-conjugated secondary antibodies were purchased from MBL International Corporation. Mitochondrial membrane potential assay kit with JC-1 (C2006) was purchased from Beyotime.
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