Anti sxbp1

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-sXBP1 is a laboratory reagent used for the detection and analysis of spliced X-box binding protein 1 (sXBP1). sXBP1 is a transcription factor that plays a key role in the unfolded protein response and endoplasmic reticulum stress pathways.

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Lab products found in correlation

Anti atf4, by Santa Cruz Biotechnology (2 mentions) Anti sxbp1, by BioLegend (2 mentions) Stat3, by ABclonal (2 mentions) Anti ire1, by Cell Signaling Technology (2 mentions) Supersignal west femto max, by Thermo Fisher Scientific (2 mentions) Supersignal west pico max chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Phospho y705 stat3, by ABclonal (2 mentions) Anti ugcg, by Abcam (2 mentions) Anti p erk, by Cell Signaling Technology (2 mentions) Complete protease inhibitor cocktail, by Roche (2 mentions) 3 amino 9 ethylcarbazole, by BioGenex (1 mentions) Anti chop, by Santa Cruz Biotechnology (1 mentions) Anti sxbp1, by Santa Cruz Biotechnology (1 mentions) Anti lc3b, by Merck Group (1 mentions) Hk330 5k, by BioGenex (1 mentions) Hk092 5k, by BioGenex (1 mentions) Hk085 5k, by BioGenex (1 mentions) Anti ire1α, by Cell Signaling Technology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti phospho p65, by Cell Signaling Technology (1 mentions)

4 protocols using anti sxbp1

Protein Extraction and Immunoblotting Assay

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Protein extraction was performed using RIPA lysis buffer (50 mM Tris HCl pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 2mM EDTA) supplemented with Complete Protease Inhibitor Cocktail (Roche) and sodium orthovanadate. After incubation on ice for 30 minutes, lysates were centrifugated 15 minutes at 4°C to remove cellular debris. Protein concentration was determined and then subjected for immunoblotting assay. The signal was visualized with Supersignal West Femto max or Supersignal West Pico max chemiluminescent substrate (Thermo Scientific) with a digital imager. The following primary antibodies were used: anti-sXBP1(D2C1F-Cell Signaling), anti-sXBP1 (6195-BioLegend), phospho-Y705 STAT3 (AP0070-Abclonal), STAT3 (A1192-Abclonal), monoclonal anti-bactin (A2228-Sigma), anti-Tubulin (AG-27B-005 Adipogen), anti-UGCG (ab124296-Abcam), anti-PERK (C33E10-Cell Signaling), anti-IRE1 (14C10-Cell Signaling), anti-ATF4 (sc-200, Santa Cruz).

Di Conza G., Tsai C.H., Gallart-Ayala H., Yu Y.R., Franco F., Zaffalon L., Xie X., Li X., Xiao Z., Raines L.N., Falquet M., Jalil A., Locasale J.W., Percipalle P., Masson D., Huang S.C., Martinon F., Ivanisevic J, & Ho P.C. (2021). Tumor-induced reshuffling of lipid composition on the ER membrane sustains macrophage survival and pro-tumorigenic activity. Nature immunology, 22(11), 1403-1415.

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Immunohistochemical Analysis of ER Stress Markers

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Lung biopsies from HP patients and control subjects and mouse lungs were FFPE. Tissues from all cases were sectioned in 5-μm-thick sections. The tissue sections were deparaffinized and rehydrated using graded ethanol (100%, 80%, and 50%) followed by water for 5 min. Endogenous peroxidase activity was eliminated using 3% H₂O₂ in methanol for 10 min and then washed in phosphate-buffered saline (PBS) 1×. Heat-induced antigen retrieval was performed in 10-mM citrate buffer (pH 6.0) for 6 min using a microwave. Lung tissues were blocked with a universal blocking solution (HK085–5K; BioGenex, Fremont, CA) for 10 min and then incubated overnight at 4C with the following primary antibodies: anti-sXBP1 (SC-7160; Santa Cruz Biotechnology, California, USA), anti-sXBP1 (83418; Cell Signaling Technology, Danvers, MA), anti-CHOP (SC-7351; Santa Cruz Biotechnology, California, USA), anti-SQSTM1/p62 (P0068; Merck, Darmstadt, Germany), and anti-LC3B (L7543; Merck). A secondary antibody cocktail followed by horseradish peroxidase–conjugated streptavidin (HK330-5K; BioGenex) was used according to the manufacturer’s instructions. The chromogenic substrate 3-Amino-9-ethyl-carbazole (HK092-5K; BioGenex) in acetate buffer containing 0.05% H₂O₂ was used, and sections were counterstained with hematoxylin.

Cabrera S., García-Vicente Á., Gutiérrez P., Sánchez A., Gaxiola M., Rodríguez-Bobadilla C., Selman M, & Pardo A. (2024). Increased ER Stress and Unfolded Protein Response Activation in Epithelial and Inflammatory Cells in Hypersensitivity Pneumonitis. Journal of Histochemistry and Cytochemistry, 72(5), 289-307.

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Protein Extraction and Immunoblotting Assay

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Western Blot Analysis of Muscle Protein Levels

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Estimation of levels of various proteins was done by performing Western blot analysis as described (Hindi and Kumar, 2016 (link); Ogura et al., 2015 (link)). TA muscle of mice were washed with sterile PBS and homogenized in lysis buffer: 50 mM Tris-Cl (pH 8.0), 200 mM NaCl, 50 mM NaF, 1 mM dithiothreitol, 1 mM sodium orthovanadate, 0.3 % IGEPAL, and protease inhibitors. Approximately 100 μg protein was resolved in each lane on 10 % SDS-polyacrylamide gels, electrotransferred onto nitrocellulose membranes, and probed with the following antibodies: anti-phospho-IRE1α (1:500; Novus, NB 100–2323), anti-IRE1α (1:500; Cell Signaling Technology, #3294), anti-sXBP-1 (1:1000; Cell Signaling Technology, #12782), anti-Pax7 (1:100; DSHB Cat# pax7, RRID:AB_528428), anti-eMyHC (1:200, DSHB Cat# F1.652 RRID:AB_528358), anti-MyoD (Santa Cruz Biotechnology sc-377460), anti-Myogenin (1:100; DSHB Cat# F5D), anti-phospho-p65 (1:500; Cell Signaling Technology, #3033), anti-p65 (1:500; Cell Signaling Technology, # 8242), anti-p100/p52 (1:500; Cell Signaling Technology, #4882), anti-Notch1 (Santa Cruz Biotechnology, #SC-6015), anti-Hes6 (Santa Cruz Biotechnology, #SC-25396), and anti-GAPDH (1:2000; Cell Signaling Technology, #2118). Antibodies were detected by chemi-luminescence. Quantitative estimation of the bands’ intensity was performed with ImageJ software (NIH).

Roy A., Tomaz da Silva M., Bhat R., Bohnert K.R., Iwawaki T, & Kumar A. (2021). The IRE1/XBP1 signaling axis promotes skeletal muscle regeneration through a cell non-autonomous mechanism. eLife, 10, e73215.

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