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application information | ||
recommended dilution | 1 : 3000 with standard ECL (WB) | |
expected | apparent MW | 29 | 29 kDa (Arabidopsis thaliana, Spinacia oleracea) | |
confirmed reactivity | Arabidopsis thaliana, Spinacia oleracea | |
predicted reactivity | dicots:Glycine max, Ricinus communis, Vitis vinifera, trees: Populus trichocarpa, monocots: Oryza sativa, Zea mays | |
not reactive in | Physcomitrella patens, Chlamydomonas reinhardtii | |
additional information | In Arabidopsis thaliana samples cross-reactivity in the region of 55-60 kDa can be detected. | |
selected references | Lintala et al. (2013). Arabidopsis tic62 trol mutant lacking thylakoid bound ferredoxin-NADP+ oxidoreductase shows distinct metabolic phenotype. Mol Plant Sep 16. Endow and Inoue (2013). Stable complex formation of thylakoidal processing peptidase and PGRL1. FEBS Lett. May 31st. |
application example 5 µg of chlorophyl a/lane from Arabidopsis thaliana wild-type thylakoids (1), Arabidopsis thaliana pgr5 mutant thylakoids (2), Arabidopsis thaliana double mutant thylakoids (3), were separated on 12% SDS-PAGE and blotted 1h to nitrocellulose membrane.Blots were blocked immediately following transfer with 5% dry milk in 1x TBS together with 0.1% Tween-20 (1x TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1:3000 for 2h at room temperature with agitation. The antibody solution was decanted and the blot washed three times, 10 min each, with 1x TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit horseradish peroxidase conjugated, from Pierce) diluted to 1:50.000 for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL detection reagent according to the manufacturer’s instructions. Exposure time was 5 min. Courtesy Paolo Pesaresi,
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