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Glutamine synthetase (leaf,root)

AS20 4426 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana, Spinacia oleracea, Zea mays
Glutamine synthetase (leaf,root) in the group Antibodies for Plant/Algal  / Nitrogen Metabolism at Agrisera AB (Antibodies for research) (AS20 4426)

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Product Information

Immunogen Purified full length, tag cleaved, recombinant Zea mays GS-1 (Glutamine synthetase, root isozyme 1) UniProt: P38559
Host Rabbit
Clonality Polyclonal
Purity Total IgG, purified on Protein A/G mix
Format Liquid at 2 mg/ml in PBS, 50% glycerol. Filter sterilized. No preservative or carrier added.
Quantity 100 µg
Storage Store at -20°C; once make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tubes.
Tested applications ELISA (ELISA), Western blot (WB)
Recommended dilution assay dependent (ELISA), 1: 1000 - 1: 5000 (WB)
Expected | apparent MW 39 kDa | 43 kDa (chloroplast isoform)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Spinacia oleracea, Zea mays
Predicted reactivity Brachypodium distachyon, Cucurbita moschata, Glycine max, Medicago truncatula, Oryza sativa, Panicum hallii, Pontederia crassipes, Populus tremula x Populus alba, Prunus persica, Raphanus sativus, Saccharum officinarum, Setaria italica, Sorghum bicolor, Triticum aestivum
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Western blot with anti-Glutamine synthetase antibodies

Recombinant root type GS isoproteins from Zea mays expressed in E.coli, negative control without recombinant GS (1), GS1-1 expressing extract (2), GS1-2 expressing extract (3), GS1-3 expressing extract (4), GS1-4  expressing extract (5), Zea mays root extrated treated with ammonia (6) were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2500 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.

The lower band in endogenous extract is ammonia induced GS.

Western blot using anti-plant glutamine synthetase antibodies
2 µg of Arabiopsis thaliana total leaf extract (1), 2 µg of Zea mays total leaf extract (2), were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.

MW of GS1 isoproteins: 38-40 kDa and this antibody reacts with both leat and root isoforms. Maize leaf contains both types. The upper band with asterisk corresponds to leaf isoform (chloroplastic), while the lower two bands correspond to root isoforms GS1-1 or/and GS1-2 (upper band)  or/and GS1-4 (lower band). 

Additional information

Additional information This antibody reactis with all glutamine synthetase isotypes from leaf and root.
No confirmed exceptions from predicted reactivity are currently known.

Related products

Background

Background Glutamine synthetase plays a role in the flow of nitrogen into nitrogenous organic compounds. There are five root types of GS isoproteis (GS1-1~ GS1-5) and one chloroplastic GS isoprotein (GS2/GLN2) are known. The sequence identity between GS1-1 and GS2 is 65%.

Product citations

Selected references Kimata-Ariga and Hase (2014). Multiple complexes of nitrogen assimilatory enzymes in spinach chloroplasts: possible mechanisms for the regulation of enzyme function. PLoS One . 2014 Oct 1;9(10):e108965. doi: 10.1371/journal.pone.0108965.
Sakaibara et al. (1996). Molecular identification and characterization of cytosolic isoforms of glutamine synthetase in maize roots. J Biol Chem. 1996 Nov 22;271(47):29561-8. doi: 10.1074/jbc.271.47.29561.

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