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GLDP | Glycine decarboxylase P protein

AS20 4370  | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Arabidopsis thaliana, Flaveria spp., Neurachne spp.,Oryza sativa, Zea mays

GLDP | Glycine decarboxylase P protein in the group Antibodies for Plant/Algal  / Mitochondria | Respiration at Agrisera AB (Antibodies for research) (AS20 4370)

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Product Information

Immunogen KLH-conjugated peptide, highly conserved in Arabidopsis thaliana GLDP1 UniProt:Q94B78, TAIR: At4g33010 and GLDP2 UniProt: O80988, TAIR: At2g26080
Host

Rabbit

Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 µl, of sterile water.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted 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 lyophilized material adhering to the cap or sides of the tubes.
Tested applications Immunolocalization (IL), Western blot (WB)
Recommended dilution 1:50 (IL), 1 : 5000 (WB)
Expected | apparent MW 113 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Flaveria spp., Neurachne spp.,Oryza sativa, Zea mays
Predicted reactivity grasses, non-succulent dicots
Species of your interest not listed? Contact us
Not reactive in Tecticornia spp., Salsola soda

Application examples

Application examples application example

Western blot using anti-plant GLDP | Glycine decarboxylase P protein antibodies

6 µg of total protein extracted freshly from Z. mays = Zea mays, A. tha= Arabidopsis thaliana, O. sat = Oryza sativa,  was freshly extracted from  leaf tissue in extraction buffer (50 mM HEPES-KOH pH 7.1, 1 mM EDTA pH 8.0, 10 mM DTT, 5 mM MgCl2, 1 mM PMSF, 20 µg/ml chymostatin, 25 µg/ml aprotinin, 1X Protease Inhibitor Cocktail VI, Plant Cell (AG Scientific)). Proteins were denatured with sample buffer (1X = 62.5 mM Tris-HCl pH 6.8, 2% [v/v] SDS, 10% [v/v] glycerol, 0.01% [w/v] bromophenol blue (Sigma), 100 mM DTT), heated to 95°C in a water bath for 3 minutes and collected at 13523 x g in a bench centrifuge. 6µg of protein was loaded per well and separated on a 12% resolving gel (0.375 M Tris-HCl pH 8.8, 12% [v/v] acrylamide, 0.1% [v/v] ammonium persulfate and 0.05% [v/v] TEMED) and 5% stacking gel (0.125 M Tris-HCl pH 6.8, 5% [v/v] acrylamide, 0.1% [v/v] SDS, 0.1% [v/v] ammonium persulfate and 0.025% [v/v] TEMED). Proteins were blotted onto nitrocellulose membrane (pore size 0.45 µm) using Trans-Blot Semi-Dry Transfer Cell (Bio-Rad), and transfer was run at 25 mA per gel stack for 1½ hours. Blots were blocked in blocking solution (5% [w/v] skim milk powder in TBST (12.5 mM Tris, 137 mM NaCl, 2.7 mM KCl, 0.1% [v/v] Tween-20 detergent)) overnight at room temperature with gentle agitation. The blot was rinsed three times quickly in TBST, then incubated in the primary antibody solution at a dilution of 1:5000 in blocking solution for 1 hour at room temperature with gentle agitation. Blots were rinsed three times quickly in TBST, then washed three times for 5 minutes in TBST at room temperature with gentle agitation. Blots were then incubated in secondary antibody solution (goat anti-rabbit IgG horse radish peroxidase (Sigma) diluted 1:3000 in blocking solution) for 1 hour at room temperature with gentle agitation. The blot was washed three times for 5 minutes in TBST at room temperature with gentle agitation, then rinsed twice in TBS (12.5 mM Tris, 137 mM NaCl, 2.7 mM KCl). Blots were developed with chemiluminescent detection reagent, following manufacture's recommendations, incubating at room temperature for 1 minute and imaged with Bio-Rad ChemiDoc system. Exposure time was 40 seconds.

Courtesy of Prof. Martha Ludwig, School of Molecular Sciences, Australia

Additional information

So far only leaf tissue has been used for immunolocalization and western blot experiments.

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Background

Background GLDP (Glycine decarboxylase P protein) catalyzes the degradation of glycine. The P protein binds the alpha-amino group of glycine through its pyridoxal phosphate cofacto. Is involved in response to cadmium ion. 

Alternative names: AtGLDP1, AtGLDP2,  Glycine dehydrogenase (aminomethyl-transferring) 1, Glycine dehydrogenase (aminomethyl-transferring) 2, Glycine cleavage system P protein 1, Glycine cleavage system P protein 2, Glycine decarboxylase 1, Glycine decarboxylase 2, Glycine decarboxylase P-protein 1, Glycine decarboxylase P-protein 2.

Product citations

Selected references Khoshravesh et al. (2020). The Evolutionary Origin of C4 Photosynthesis in the Grass Subtribe Neurachninae.Plant Physiol. 2020 Jan;182(1):566-583. doi: 10.1104/pp.19.00925.

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