EPSP synthase | 3-phosphoshikimate 1-carboxyvinyltransferase (chloroplastic, Arabidopsis thaliana)

AS14 2782 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, di and monocots

Product Information

Immunogen

KLH-conjugated peptide, derived from Arabidopsis thaliana EPSP synthase, UniProt: P05466 TAIR:At2g45300

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 Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

55 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Arachis hypogaea, Cannabis sativa, Coffea arabica, Cucumis sativus, Glycine max, Glycine soja, Malus domestica, Nicotiana tabacum, Zea mays, Vitis vinifera
Species of your interest not listed? Contact us

Application examples

Application examples application example

0.5 µg of purified Arabidopsis thaliana EPSPS was separated on 10 % SDS-PAGE and blotted 2h to nitrocellulose Hibond ECLmembrane (GE Healthcare) using semi-dry or tank transfer. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody in TBS-T with (0.1 % Tween 20) at a dilution of 1: 1 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 2 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1: 20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Protein detection was done by Image Quant LAS4000 (GE-Health Care) digital imager system with exposure time was of 5 minutes.

Recombinant Arabidopsis thaliana EPSPS protein (aka 3-phosphoshikimate 1-carboxyvinyltransferase) used in the assay was identified by LCMS.

Courtesy of Dr. Altanbadralt Sharkhuu, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia

50 µg of total protein from shoots of 18 d old Arabidopsis thaliana were extracted with buffer (125 mM Tris-HCl ph6.8; 12% SDS; 10% glycerol, 22% b-mercaptoethanol; 0.001% bromophenol blue) were separated on % SDS-PAGE and blotted 3h to nitrocellulose membrane using tank transfer. Blots were blocked with TBST with 5% skim milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody (Rabbit-EPSPS) at a dilution of 1: 1 000 for over night at 4 C 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 RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Detection was done by Image Quant LAS 4000 (GE-Health Care). Exposure time was 2 min.

Courtesy of Dr. Altanbadralt Sharkhuu, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia

Additional information

Related products

AS16 3119 | Anti-EPSP synthase | 3-phosphoshikimate 1-carboxyvinyltransferase, chloroplastic, rabbit antibodies

Recommended secondary antibody for ECL detection

Plant protein extraction buffer

Background

Background EPSP synthase (3-phosphoshikimate 1-carboxyvinyltransferase, chloroplastic) is an enzyme (EC:2.5.1.19)which is involved in glyphosate metabolic process. Localized in chloroplast stroma. Catalyzes the transfer of the enolpyruvyl moiety of phosphoenolpyruvate (PEP) to the 5-hydroxyl of shikimate-3-phosphate (S3P) to produce enolpyruvyl shikimate-3-phosphate and inorganic phosphate.

Alternative name: 5-enolpyruvylshikimate-3-phosphate synthase.