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GlnA | Glutamine synthetase

265 €
Buy 2 items of this product for 198 €/each
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AS01 018  |  clonality: polyclonal   | host: hen  |  reactivity: [global antibody] for bacteria

PRODUCT INFORMATION IN PDF

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AS01 018

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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 Hen
Clonality Polyclonal
Clone
Purity Total IgY
Format Liquid in PBS pH 8.0, 0.02% sodium azide
Quantity 50 ĩl (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 control

Recommended secondary antibody

collection 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, Arthropsira sp. 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

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) 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.

western blot using anti-GlnA antibodies on cyanobacterial sampls

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 manufacturer's instructions. Images of the blots were obtained using a CCD imager and Quantity One software. Exposure time was 15 seconds.

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