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application information | ||
recommended dilution | 1 : 2 000 with standard ECL (WB) | |
expected | apparent MW | 120 | ~130 for Zea mays | |
confirmed reactivity | Zea mays | |
predicted reactivity | monocots:Saccharum officinarum, Triticum aestivum and Oryza sativa | |
not reactive in | Hordeum vulgare | |
additional information | to be added when available | |
selected references | Sparks et al. (2001) A Appl Biol 138: 33-45.Potential for manipulating carbon metabolism in wheat |
application example 10 µg of total leaf protein from (1) A.thaliana, (3) Zea mays and (4) Hordeum vulgare extracted with PEB (AS08 300) as well as 10 µg cytosolic protein from (2) A.thaliana were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1.5h (30V) to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-SPS (AS06 0185, 1:2000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated, Abcam) in TBS-T containing 2% low fat milk powder. Antibody incubations where followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with standard ECL (GE Healthcare) using a Fuji LAS-3000 CCD (90s, high sensitivity). |