Mouse monoclonal antibodies for anti-6X his-tag (Abcam - ab18184) (1:1,000), HA (Biolegend 901501) (1:1,000), α-tubulin (Clone 12G10 from Developmental Studies Hybridoma Bank, University of Iowa) (1:5,000) and viral envelope (Merck-Millipore MAB10216) (1:1,000) were used. Rabbit polyclonal antibodies for GAPDH (1:5,000) were from Cell Signaling Technology (CST 2118). Rabbit polyclonal dengue-capsid antibody was used for immunoblotting (1:1,000) (Agrawal et al., 2013 (link)). Rabbit polyclonal DDX3X (R648) antibody (1:2,000 for western blotting and 1:1,000 for immunofluorescence) has been described before (Angus et al., 2010 (link)). Anti-rabbit (Merck-Millipore) or anti-mouse (Sigma) IgG secondary antibodies raised in goat and conjugated with horseradish peroxidase (HRP) (1:2,000) were used. HRP-conjugated secondary antibodies specific to IgG-light chain (Jackson's Immunoresearch Lab) were used (1:10,000) for probing blots of immunoprecipitation.
Cst 2118
Manufactured by Cell Signaling Technology
Sourced in United States
The CST 2118 is a laboratory equipment product offered by Cell Signaling Technology. It is a core component for conducting biological research and experiments. The primary function of this equipment is to facilitate the analysis and study of cell signaling pathways and mechanisms within a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Mab10216, by Merck Group (1 mentions)
Horseradish peroxidase linked anti rabbit igg, by Cell Signaling Technology (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Sc 47778, by Santa Cruz Biotechnology (1 mentions)
Chemidox xrs, by Bio-Rad (1 mentions)
Ab125066, by Abcam (1 mentions)
Ab175186, by Abcam (1 mentions)
Cst7074, by Cell Signaling Technology (1 mentions)
2 protocols using cst 2118
1
Antibody Validation for Protein Detection
2
Ferroptosis Marker Protein Analysis
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The neurons were lysed with lysis buffer (Beyotime, Nantong, China), and the proteins were separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred onto polyvinylidene difluoride membranes. The membranes were blocked in 5% nonfat milk at room temperature for 1 hour, and thereafter incubated with primary antibodies [anti-GPX4 (1:5000; ab125066, rabbit, Abcam), anti-xCT (1:1000; ab175186, rabbit, Abcam), anti-β-actin (1:5000; sc47778, mouse, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-GAPDH (1:2000; CST2118, rabbit, Cell Signaling Technology, Boston, MA, USA)] overnight. GPX4 and xCT are both key ferroptosis markers. The blots were then incubated with secondary antibodies (horseradish peroxidase-linked anti-mouse IgG (CST7076, 1:3000; Cell Signaling Technology) and horseradish peroxidase-linked anti-rabbit IgG (CST7074, 1:3000; Cell Signaling Technology) at room temperature for 1 hour. Finally, the blots were visualized using an enhanced chemiluminescence system (ChemiDox XRS; Bio-Rad, Hercules, CA, USA). All grayscale values were normalized to that of β-actin or GAPDH using ImageJ software (National Institutes of Health).
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