product information Background Glutamine synthetase (EC=6.3.1.2) is the key enzyme in the incorporation of mineral nitrogen into glutamine. Activity of this enzyme is controlled by adenylarion under conditions of abundant glutamine. Immunogen KLH-conjugated synthetic peptide derived from available bacterial GlnA sequences with perfect conservation in alpha, beta, gamma Proteobacteria, Enterobacteria, Thermotogales, Low GC Gram+, Cyanobacteria (except weak conservation with Trichodesmium thiebautii) including Synechocystis PCC 6803 Q59981 Host Chicken Clonality Polyclonal Clone Purity Total IgY Format Liquid in PBS pH 8.0, 0.02% sodium azide Quantity 50 µl (16 mg/ml) Reconstitution Storage store at 4°C; make aliquots to avoid working with a stock. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from liquid material adhering to the cap or sides of the tubes. Tested applications western blot (WB) Related products AS01 018S | GlnA protein standard for a quantitative western blot or as a positive controlRecommended secondary antibodycollection of antibodies to nitrogen metabolism Algal protein extraction buffer Secondary antibodies Additional information Peptide target used to elicit this antibody has a weak, sporadic conservation with Glutamine Synthetase to III, antibody not expected to detect this enzyme. Weak conservation with some Glutaminyl-tRNA synthetase (Glutamine--tRNA ligase) (GLNRS), but this antibody is not expected to detect this enzyme.

application information
Recommended dilution		1:5000 with standard ECL (WB)
Expected | apparent MW		53 kDa
Confirmed reactivity		Synechococcus sp. strain PCC 7942, Synechocystis sp. strain PCC 6803, Trichodesmium IMS
Predicted reactivity		alpha, beta, gamma proteobacteria, enterobacteria, thermotogales, euryarchaeotes, crenarchaeotes, Arthropsirasp. PCC 8005
Not reactive in		diatoms, eukaryotic GlnA
Additional information
Selected references		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.

application example

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 manufacturers instructions. Images of the blots were obtained using a CCD imager nd Quantity One software  Exposure time was 10 seconds.

Total protein (1.5 μg) from Synechococcus sp. strain PCC 7942 (1) and Synechocystis sp. strain PCC 6803 (2) and GlnA recombinant protein standard (AS09 018S), 600, 400 and 200 fmol (3-5) were separated on a 4-12% Bolt gel (Thermo-Fisher) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL blocking agent (GE Life Sciences) 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 GlnA primary anYbody (AS01 018) diluted to 1:20 000 in 2% ECL blocking solulution for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly three times, 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-chicken IgY horseradish peroxidase conjugated, AS10 1489) diluted to 1:20 000 in 2% ECL blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Select detection reagent (GE Life Sciences) according the manufacturers instructions. Images of the blots were obtained using a CCD imager and Quantity One software. Exposure time was 15 seconds.

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