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Bioss p ire1α
P Ire1α, supplied by Bioss, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ire1a+(ser726)+polyclonal+antibody/IRE1a+(Ser726)+Polyclonal+Antibody/pm35696049-55-9-11
Average 92 stars, based on 8 article reviews

p ire1α - by Bioz Stars, 2026-09

92/100 stars

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Western Blot:

Article Title: The IRE1/JNK signaling pathway regulates inflammation cytokines and production of glomerular extracellular matrix in the acute kidney injury to chronic kidney disease transition.
Article Snippet: Background The transition from acute kidney injury (AKI) to chronic kidney disease (CKD) is extremely complex.. Incomplete renal tubule repair, inflammation, and endoplasmic reticulum (ER) stress all play major roles.. AKI activates ER stress, and the sensor protein inositol-requiring kinase-1 (IRE1) mediates inflammation by promoting the phosphorylation of C-jun NH2-terminal kinase (JNK).

Incubation:

Article Title: Mesencephalic astrocyte-derived neurotrophic factor (MANF) protects against Aβ toxicity via attenuating Aβ-induced endoplasmic reticulum stress
Article Snippet: .. The proteins were transferred to PVDF membranes and blocked in 5% nonfat milk at room temperature for 1 h, then incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MANF antibody (1: 1000, Abcam, ab67271), rabbit anti-BiP antibody (1:1000, proteintech, 11587-1-ap), rabbit anti-CHOP antibody (1:1000, proteintech, 15204-1-AP), rabbit anti-phospho-eIF2α antibody (1:1000, CST, 3398 s), rabbit anti-phospho-IRE1 antibody (1:1000, Bioss, bs-4308R), rabbit anti-XBP1s antibody (1:1000, Biolegend, 619502), mouse anti-ATF4 antibody (1:1000, CST, 11815 s), rabbit anti-ATF6 antibody (1:1000, Proteintech, 24169-1-AP), rabbit anti-cleaved-caspase 3 antibody (1: 1000, CST, 9664S), mouse anti-α-tubulin (1: 1000, sigma, t6199), and rabbit-anti-GAPDH (1: 1000, Elabscience, E-AB-20059). .. After washing several times in TBST, membranes were incubated with horseradish peroxidase-conjugated secondary IgG for 1 h. Blots were developed using the enhanced chemiluminescence kit (Amersham Biosciences, NJ, USA).

Article Title: Xanthatin induces glioma cell apoptosis and inhibits tumor growth via activating endoplasmic reticulum stress-dependent CHOP pathway
Article Snippet: .. After electrophoresis and transfer, the PVDF membranes were blocked with 5% nonfat milk powder in TBST buffer and then incubated with primary antibodies against the following proteins at 4 °C overnight: glucose-regulated protein 78 (GRP78) (1:1000, Proteintech, 11587-1-AP), CHOP (1:1000, Santa Cruz Biotechnology, sc-793), phospho-eIF2α (1:1000, CST, 3398 s), phospho-IRE1α (1:1000, Bioss, bs-4308R), X-box binding protein-1 (XBP1s) (1:1000, BioLegend, 619502), activating factor 4 (ATF4) (1:1000, CST, 11815 s), activating transcription factor 6 (ATF6) (1:1000, Proteintech, 24169-1-AP), cleaved caspase-3 (1:1000, CST, 9654 s), Bcl-2 (1:1000, Bioss, bs-0032R), Bax (1:500, Immunoway, YT0456), tubulin (1:1000, Sigma, t6199), and GAPDH (1:1000, BioLegend, 619502). .. The membranes were incubated with the corresponding horseradish peroxidase-conjugated secondary IgG for 1 h. Blots were developed using an enhanced chemiluminescence kit (Amersham Biosciences, NJ, USA).

Electrophoresis:

Binding Assay:

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Recombination human MANF protein (rhMANF) protects against Aβ 1–42 -induced cell toxicity. a The dose-dependent effect of rhMANF on cell viability in the presence of Aβ 1–42 exposure. SH-SY5Y cells were pretreated with different concentrations of rhMANF protein (0.5, 1, and 2.0 mg/ml) for 4 h prior to the Aβ 1–42 (10 μM) treatment for additional 24 h and processed for the MTT assay. * P < 0.05, compared with the cells only treated with Aβ 1–42 . b The time-course of rhMANF on cell viability in the presence of Aβ 1–42 . SH-SY5Y were pretreated with rhMANF (2.0 mg/ml) for 4 h following the Aβ 1–42 (10 μM) treatment for indicated times and processed for the MTT assay. * P < 0.05, compared with the only Aβ 1–42 -treated cells at indicated time points. c The protein levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 were determined in SH-SY5Y cells treated with Aβ 1–42 (10 μM) for 24 h with or without rhMANF. GAPDH was used as a loading control. d The densitometric quantitation of indicated proteins normalized to GAPDH levels in c. * P < 0.05, ** P < 0.01, compared with the cells only treated with Aβ 1–42 . # P < 0.05, ## P < 0.01, ### P < 0.001, compared with control group. All the quantitative data were presented as mean + SD of at least three independent experiments. C - caspase - 3 cleaved caspase-3

Journal: Journal of Neuroinflammation

Article Title: Mesencephalic astrocyte-derived neurotrophic factor (MANF) protects against Aβ toxicity via attenuating Aβ-induced endoplasmic reticulum stress

doi: 10.1186/s12974-019-1429-0

Figure Lengend Snippet: Recombination human MANF protein (rhMANF) protects against Aβ 1–42 -induced cell toxicity. a The dose-dependent effect of rhMANF on cell viability in the presence of Aβ 1–42 exposure. SH-SY5Y cells were pretreated with different concentrations of rhMANF protein (0.5, 1, and 2.0 mg/ml) for 4 h prior to the Aβ 1–42 (10 μM) treatment for additional 24 h and processed for the MTT assay. * P < 0.05, compared with the cells only treated with Aβ 1–42 . b The time-course of rhMANF on cell viability in the presence of Aβ 1–42 . SH-SY5Y were pretreated with rhMANF (2.0 mg/ml) for 4 h following the Aβ 1–42 (10 μM) treatment for indicated times and processed for the MTT assay. * P < 0.05, compared with the only Aβ 1–42 -treated cells at indicated time points. c The protein levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 were determined in SH-SY5Y cells treated with Aβ 1–42 (10 μM) for 24 h with or without rhMANF. GAPDH was used as a loading control. d The densitometric quantitation of indicated proteins normalized to GAPDH levels in c. * P < 0.05, ** P < 0.01, compared with the cells only treated with Aβ 1–42 . # P < 0.05, ## P < 0.01, ### P < 0.001, compared with control group. All the quantitative data were presented as mean + SD of at least three independent experiments. C - caspase - 3 cleaved caspase-3

Article Snippet: The proteins were transferred to PVDF membranes and blocked in 5% nonfat milk at room temperature for 1 h, then incubated at 4 °C overnight with the following primary antibodies: rabbit anti-MANF antibody (1: 1000, Abcam, ab67271), rabbit anti-BiP antibody (1:1000, proteintech, 11587-1-ap), rabbit anti-CHOP antibody (1:1000, proteintech, 15204-1-AP), rabbit anti-phospho-eIF2α antibody (1:1000, CST, 3398 s), rabbit anti-phospho-IRE1 antibody (1:1000, Bioss, bs-4308R), rabbit anti-XBP1s antibody (1:1000, Biolegend, 619502), mouse anti-ATF4 antibody (1:1000, CST, 11815 s), rabbit anti-ATF6 antibody (1:1000, Proteintech, 24169-1-AP), rabbit anti-cleaved-caspase 3 antibody (1: 1000, CST, 9664S), mouse anti-α-tubulin (1: 1000, sigma, t6199), and rabbit-anti-GAPDH (1: 1000, Elabscience, E-AB-20059).

Techniques: MTT Assay, Quantitation Assay

MANF protects against Aβ 1–42 toxicity via inhibiting ER stress. a Effect of MANF overexpression on the levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 in Aβ 1–42 -treated cells. N2a cells were transfected with pcDNA3.1-MANF-Flag plasmid and its empty vector before Aβ 1–42 (10 μM) treatment for 24 h or TM (2.5 μg/ml) treatment for 12 h and processed for WB. b Effect of MANF knockdown on the levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 in Aβ 1–42 -treated cells. N2a cells were transfected with NC-siRNA or MANF-siRNA before Aβ 1–42 (10 μM) treatment for 24 h or TM (2.5 μg/ml) treatment for 12 h, respectively, then processed for WB. c Quantitation of proteins normalized to α-tubulin levels in a. d Quantitation of proteins normalized to α-tubulin levels in b . All the quantitative data were presented as mean ± SD of at least three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.01 . TM tunicamycin, C - caspase - 3 cleaved caspase-3

Journal: Journal of Neuroinflammation

Article Title: Mesencephalic astrocyte-derived neurotrophic factor (MANF) protects against Aβ toxicity via attenuating Aβ-induced endoplasmic reticulum stress

doi: 10.1186/s12974-019-1429-0

Figure Lengend Snippet: MANF protects against Aβ 1–42 toxicity via inhibiting ER stress. a Effect of MANF overexpression on the levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 in Aβ 1–42 -treated cells. N2a cells were transfected with pcDNA3.1-MANF-Flag plasmid and its empty vector before Aβ 1–42 (10 μM) treatment for 24 h or TM (2.5 μg/ml) treatment for 12 h and processed for WB. b Effect of MANF knockdown on the levels of BiP, ATF6, phospho-IRE1, XBP1s, phospho-eIF2α, ATF4, CHOP, and cleaved caspase-3 in Aβ 1–42 -treated cells. N2a cells were transfected with NC-siRNA or MANF-siRNA before Aβ 1–42 (10 μM) treatment for 24 h or TM (2.5 μg/ml) treatment for 12 h, respectively, then processed for WB. c Quantitation of proteins normalized to α-tubulin levels in a. d Quantitation of proteins normalized to α-tubulin levels in b . All the quantitative data were presented as mean ± SD of at least three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.01 . TM tunicamycin, C - caspase - 3 cleaved caspase-3

Techniques: Over Expression, Transfection, Plasmid Preparation, Quantitation Assay

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