WB: Assay dependent. ICC/IF: 1 µg/ml (1/500 dilution) -5 µg/ml (1/100 dilution). IP: 1-5µg/sample. ELISA: Assay dependent. IHC: 1 µg/ml (1/500 dilution) -5 µg/ml (1/100 dilution). Target : SOD1 mutants. Target : SOD1 mutants under native condition ; Wild-type SOD1 under the zinc-deficient ER-stress. GTX57211 and GTX57212 could recognize not only mutants but also denatured SOD1. Please keep non-denatured condition and avoid to use high concentration of SDS or the other denaturing compounds.. Target : SOD1 mutants. *Optimal dilutions/concentrations should be determined by the researcher.Not tested in other applications.
ICC/IF analysis of Flag-tagged SOD1 wild type or G93A mutant-expressing HEK293 cells using GTX57211 SOD1 (mutant) antibody [MS785], GTX57212 SOD1 (mutant) antibody [MS27] or MS785/MS27 cocktail. Fixation : 4% PFA for 10 min at RT.
Permeabilization : 0.2% Triton X-100 for 5 min
Dilution : 1μg/ml (12 hours at 4ºC)
IP analysis of HEK293 cells cultured in the presence and absence of 10μM TPEN (a potent zinc-specific chelator, for 8 hours) using GTX57211 SOD1 (mutant) antibody [MS785]. Under the zinc-deficient ER-stress, these antibodies could recognize endogenous SOD1 wild type with mutant like conformation. Antibody amount : 1-5 μg
IP analysis of SOD1 wild type or mutants-expressing HEK293 cells using GTX57211 SOD1 (mutant) antibody [MS785], GTX57212 SOD1 (mutant) antibody [MS27], or MS785/MS27 cocktail. Neither MS785 nor MS27 single detected some specific mutants which have the mutation on the antibody’s epitope. MS785/MS27 cocktail overcame this problem. Validated SOD1 mutants were listed in “Appendix”.
Antibody amount : MS27 (GTX57212): 2μg, MS785 (GTX57211): 5μg, or MS27/MS785 cocktail: 1μg
IP lysis buffer: 1% Triton X100/TBS buffer.
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