To determine the metal content in yeast mitochondria, cells transformed with either the empty pUG23 vector or pUG23 plasmid carrying CsMIT1 or CsMIT2 were grown for 48 h in SC/Glu-His medium and then for 12 h in the same medium supplemented with 100 µM BPS. Mitochondrial fractions were prepared as previously described [58 (link)]. The presence of CsMIT1 and CsMIT2 in the mitochondrial fraction was confirmed by Western blot analysis using antibodies against GFP (1:5000, Roche, Basel, Switzerland).
Antibodies against gfp
Manufactured by Roche
Sourced in Switzerland
Antibodies against GFP are laboratory reagents that specifically bind to the green fluorescent protein (GFP) molecule. They are commonly used in various techniques, such as Western blotting, immunoprecipitation, and immunocytochemistry, to detect and study the expression, localization, and interactions of GFP-tagged proteins.
Automatically generated - may contain errors
4 protocols using antibodies against gfp
1
Metal Content in Yeast Mitochondria
2
GFP Antibodies Western Blotting Protocol
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Antibodies against GFP were obtained from Roche Applied Science (ref: 11814460001) and used according to the manufacturer’s instructions. Western blotting was performed as described previously (6 (link)) with 30 µg of total proteins loaded per sample.
3
Detection of Transcription Factor and RBP
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c-Myb was detected with Myb-specific monoclonal antibodies 5E11 (35 (link)). Pdcd4 was detected with a rabbit antiserum against the N-terminus of human Pdcd4 (12 (link)). Antibodies against GFP (Roche Diagnostics, Mannheim, Germany), eIF4A, eIF4G, eIF4E, PABP (Cell Signalling Technology), Y14 (Santa Cruz Biotechnology) and the Flag, Myc and His-tag were obtained from commercial suppliers. Antiserum against Drosophila PABP was kindly provided by E. Izaurralde (Tübingen, Germany).
4
Alr3234::GFP Western Blot Analysis in E. coli and Nostoc
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For Alr3234::GFP Western blot analysis in E. coli , cells from stationary-phase cultures were harvested and resuspended in SDS-PAGE loading buffer. Proteins were fractionated in 10% SDS–PAGE. Antibodies against GFP (Roche) and E. coli GroEL (Sigma-Aldrich) were used. For Alr3234::GFP Western blot analysis in Nostoc, soluble fractions were prepared as previously described (40 (link)). Forty-microgram amounts of protein were fractionated on 10% SDS–PAGE gels, and Antibodies against GFP were used. Ponceau staining was used as a loading control. The ECL plus immunoblotting system (GE Healthcare) was used to detect the different primary antibodies using anti-rabbit (Sigma-Aldrich) or anti-mouse (Bio-Rad) horseradish peroxidase-conjugated secondary antibodies.
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