HydA2 | Iron-hydrogenase HydA2
AS09 600 | clonality: polyclonal | host: rabbit | reactivity: Chlamydomonas reinhardtii
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1 : 5000 with standard ECL (WB)
|Expected | apparent MW||
53.7 | 47.3 kDa (after a signal peptide is cleaved)
|Confirmed reactivity||Chlamydomonas reinhardtii, recombinant HydA2 expressed in E.coli
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
HydA2 (505aa) has a calculated MW of 53.7 kDa, but this is including the signal peptide, which gets cleaved off. The protein without TP can only be estimated, since the cleavage site is known only from in silico analysis. It has a calculated MW of 47.3 kDa and should run in the gel also according to its size.
HydA1 (497aa) has a calculated MW of 53.1 kDa, but this is including the signal peptide, which gets cleaved off. The protein without TP has a calculated MW of 47.5 kDa and runs according to its size at about 48 kDa
To be added when available
50 ng of purified protein (HydA1 and HydA2) were separated on 10% SDS-PAGE and blotted 25 min to PVDF membrane. Filters were blocked 1h with 3% low-fat milk powder in PBS-T (0.1% TWEEN 20) and probed with anti-HydA2 (AS09 600, 1:5000, over night at 4°C) and secondary anti-rabbit (1:10 000, 1 h) antibody (HRP conjugated, manufacture Pierce) in PBS-T containing 3% low fat milk powder. Antibody incubations were followed by washings in PBS-T (10, +10min and PBS (+5, +5 min). All washing steps were performed at RT with agitation. Signal was detected with ECL (Millipore) using CCD camer. Exposure time was 20 min.
Courtesy Dr. Thomas Happe
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