GlnA | glutamine synthetase

320 €

AS01 018 | clonality: polyclonal | host: hen | reactivity: [global antibody] for bacteria

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background

Glutamine synthetase is the key enzyme in the incorporation of mineral nitrogen into 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

antibody format

hen

polyclonal

total IgY in PBS pH 8.0+ 0.02% sodium azide, conc.16 µg/µl

liquid

quantity

50 µl

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

GlnA protein standard for quantitation and positive control

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 antibody 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. 7942, Synechocystis sp. 6803
predicted reactivity		alpha, beta, gamma proteobacteria, enterobacteria, thermotogales, euryarchaeotes, crenarchaeotes, Arthropsira sp. PCC 8005
not reactive in		diatoms, eukaryotic GlnA
additional information		n.a.
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

western blot using anti- GlnA antibodies

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, from Abcam) 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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