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product information
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| background |
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Glutamine synthetase is the key enzyme in the incorporation of mineral nitrogen into glutamine. |
| immunogen |
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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 |
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hen |
polyclonal |
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total IgY in PBS pH 8.0+ 0.02% sodium azide, conc.16 µg/µl |
liquid |
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| quantity |
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| storage |
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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 |
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western blot (WB) |
| related products |
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GlnA protein standard for quantitation and positive control |
| additional information |
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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
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| recommended dilution |
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1:5000 with standard ECL (WB) |
| expected | apparent MW |
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53 kDa |
| confirmed reactivity |
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Synechococcus sp. 7942, Synechocystis sp. 6803 |
| predicted reactivity |
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alpha, beta, gamma proteobacteria, enterobacteria, thermotogales, euryarchaeotes, crenarchaeotes, Arthropsira sp. PCC 8005 |
| not reactive in |
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eukaryotic GlnA |
| additional information |
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n.a. |
| selected references |
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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, 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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