Western blot analyses with whole-cell RIPA buffer protein lysates were separated by SDS-PAGE and blotted onto Hybond-C-Extra nitrocellulose membranes (Amersham Bioscience, Little Chalfont, UK) or onto Trans-Blot Turbo Mini PVDF membranes (BioRad). Following primary antibodies were used after blocking: anti-HIF-1α (1:500, rabbit polyclonal, Millipore, CA) anti-HIF-2α (1:500, rabbit polyclonal Abcam), anti-c-Myc (1:100, mouse monoclonal Santa Cruz), anti-GLS (1:500, rabbit polyclonal, Proteintech), anti-SDHA (1:2,000, mouse monoclonal Abcam) and anti-β-Actin (1:500, mouse monoclonal Santa Cruz). The proteins were detected by using HRP-conjugated secondary antibodies; anti-mouse IgG (1:5,000, GE healthcare, Buckinghamshire, UK) and anti-rabbit IgG (1:5,000, GE healthcare) and EZ-ECL chemiluminescence detection kit (Biological Industries, Israel).
Anti gls
Manufactured by Proteintech
Sourced in United States
Anti-GLS is a laboratory reagent used to detect and quantify the presence of glutaminase (GLS) in biological samples. It functions by specifically binding to and capturing GLS proteins, enabling their identification and measurement.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit cy5, by Thermo Fisher Scientific (2 mentions)
Goat serum, by Thermo Fisher Scientific (2 mentions)
Anti neun, by Abcam (2 mentions)
Hybond c extra nitrocellulose membrane, by Cytiva (1 mentions)
Trans blot turbo mini pvdf membrane, by Bio-Rad (1 mentions)
Anti hif 1α, by Merck Group (1 mentions)
Anti sdha, by Abcam (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Anti hif 2α, by Abcam (1 mentions)
Anti c myc, by Santa Cruz Biotechnology (1 mentions)
Ez ecl chemiluminescence detection kit, by Sartorius (1 mentions)
Hybond, by Cytiva (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti vglut1, by Thermo Fisher Scientific (1 mentions)
4 protocols using anti gls
1
Western Blot Protocol for Hypoxia Regulators
2
Perfusion and Immunostaining Protocol for Brain Tissue
3
Perfusion and Immunostaining Protocol for Brain Tissue
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After fMRI imaging, the animals were perfused with saline followed by 4% PFA solution. The brain was taken out carefully and stored in solution with 4% paraformaldehyde and 20% sucrose. After fixation, the brain was sliced into 60 μm slices. Fluorescent expression in the injection site was evaluated under microscopy. Immunofluorescent staining was performed to further confirm the AAV expression. After being washed with phosphate buffered saline (PBS), sections were treated with 1% Triton X-100 for 15 minutes followed by 1-hour blocking with 10% goat serum (ThermoFisher Scientific) at room temperature, then incubated with primary antibodies (1:50, anti-NeuN, Abcam; 1:20, anti-GLS, Proteintech) at 4 degree overnight, and followed by secondary antibody (1:200, goat-anti-rabbit-Cy5 and goat-anti-mouse-488, ThermoFisher Scientific) incubation for 1 hour at room temperature. Lastly, immunostained sections were stained with DAPI and mounted for fluorescent confocal microscopy.
4
Metabolic Pathway Profiling in Methamphetamine and Amphetamine Exposure
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Methamphetamine hydrochloride (METH), Amphetamine hydrochloride (AMPH), all-trans-retinoic acid (ATRA) were obtained from Sigma-Aldrich Chemicals (USA). The primary antibodies used were as follows: anti-P5CS (cat# NBP1-83324) was purchased from Novus Biologicals (USA), anti-PYCR1 (cat# 13108-1-AP, anti-PYCR2 human (cat# 55060-1-AP, anti-PYCR2 mouse (cat #17146-1-AP, anti-P5CDH (cat# 11604-1-AP), anti-GLS (cat# 12855-1-AP) were purchased from Proteintech (USA), anti-POX IgG was a gift from Dr. James Phang (NCI-Frederick), anti-vGLUT1 (cat# 48-2400) was purchased from Thermo Fisher (USA), and anti-β-actin was obtained from Sigma-Aldrich Chemicals (USA). The secondary antibodies used were goat anti-rabbit or goat anti-mouse purchased from BioRad laboratories (USA).
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