GlnA | Glutamine synthetase
AS01 018 | clonality: polyclonal | host: hen | reactivity: [global antibody] for bacteria
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1:5000 with standard ECL (WB)
|Expected | apparent MW||
Synechococcus sp. 7942, Synechocystis sp. 6803
alpha, beta, gamma proteobacteria, enterobacteria, thermotogales, euryarchaeotes, crenarchaeotes, Arthropsira sp. PCC 8005
|Not reactive in||
diatoms, eukaryotic GlnA
Brown et al. (2008). Flux capacities and acclimation costs in Trichodesmium from the Gulf of Mexico. Marine Biol. 154:413-422.
Burns et al. (2006). Inorganic carbon repletion constrains steady-state light acclimation in the cyanobacterium Synechococcus elongatus. J. Phycol. 42:610-621.
3 µg of total protein from Trichodesmium IMS 101 extracted with PEB (AS08 300) (1-8) and GlnA protein standard 0.3, 0.15, 0.07 pmol (9-11) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgY horseradish peroxidase conjugated) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according the manufacturer's instructions. Images of the blots were obtained using a CCD imager nd Quantity One software Exposure time was 10 seconds.
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