Protein was extracted from whole cells by lysis in RIPA buffer (Pierce/Thermo Scientific) supplemented with protease and phosphate inhibitor cocktail (Pierce/Thermo Scientific). Lysates were cleared using centrifugation, resolved on Bolt 4-12% bis-tris pre-cast gels (Invitrogen, Life Technologies) and transferred to nitrocellulose membranes. Primary antibodies: MTAP (Proteintech, 11475-1-AP), AMD1 (Proteintech, 11052-1-AP), ODC1 (Proteintech, 17003-1-AP), GPX4 (Abcam, Ab125066), AHCY (Abcam, ab134966, EPR9261), CBS (Abcam, ab140600, EPR8579), CSE (Abcam, ab189916, EPR15468) and actin (EMD Millipore, MAB1501, clone C4) as loading control on the same blots. Secondary antibodies: IRDye800CW anti-mouse (925-32212), IRDye680RD anti-mouse (925-68072), IRDye800CW anti-rabbit (925-32213), IRDye680RD anti-rabbit (925-68073). Protein bands were detected and quantified using a Li-Cor Odyssey Fc infrared scanner and Image Studio (v5.2) software (Li-Cor Biosciences).
Bolt 4 12 bis tris pre cast gels
Manufactured by Thermo Fisher Scientific
Bolt 4-12% bis-tris pre-cast gels are a type of polyacrylamide gel used for electrophoresis. The gels have a bis-tris buffer system and a 4-12% gradient, which allows for the separation and analysis of a wide range of protein sizes.
Automatically generated - may contain errors
Lab products found in correlation
Ab134966, by Abcam (2 mentions)
Ab189916, by Merck Group (2 mentions)
Ab140600, by Abcam (2 mentions)
Ab125066, by Abcam (2 mentions)
Image studio v5, by LI COR (2 mentions)
Odyssey fc infrared scanner, by LI COR (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Protease and phosphate inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
2 protocols using bolt 4 12 bis tris pre cast gels
1
Western Blot Analysis of Metabolic Enzymes
2
Western Blot Analysis of Metabolic Enzymes
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Protein was extracted from whole cells by lysis in RIPA buffer (Pierce/Thermo Scientific) supplemented with protease and phosphate inhibitor cocktail (Pierce/Thermo Scientific). Lysates were cleared using centrifugation, resolved on Bolt 4-12% bis-tris pre-cast gels (Invitrogen, Life Technologies) and transferred to nitrocellulose membranes. Primary antibodies: MTAP (Proteintech, 11475-1-AP), AMD1 (Proteintech, 11052-1-AP), ODC1 (Proteintech, 17003-1-AP), GPX4 (Abcam, Ab125066), AHCY (Abcam, ab134966, EPR9261), CBS (Abcam, ab140600, EPR8579), CSE (Abcam, ab189916, EPR15468) and actin (EMD Millipore, MAB1501, clone C4) as loading control on the same blots. Secondary antibodies: IRDye800CW anti-mouse (925-32212), IRDye680RD anti-mouse (925-68072), IRDye800CW anti-rabbit (925-32213), IRDye680RD anti-rabbit (925-68073). Protein bands were detected and quantified using a Li-Cor Odyssey Fc infrared scanner and Image Studio (v5.2) software (Li-Cor Biosciences).
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