PPDK | Pyruvate orthophosphate dikinase

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AS13 2647 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana, Zea mays


9 st
Item No:
AS13 2647

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product information
Background PPDK (Pyruvate, phosphate dikinase 1) is involved in formation of phosphoenolpyruvate and activated by light-induced dephosphorylation. Inhibited by dark-induced phosphorylation. PPDK is a low-abundance enzyme in C3 plants while it is a key enzyme of C4 photosynthesis.

Purified recomibinant enzyme consisting of residues 72-947 of Zea mays, UniProt: P11155

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum
Format Lyophilized
Quantity 20 ĩg
Reconstitution For reconstitution add 200 ĩl of sterile water in 40% glycerol to a final protein concentration of 100 ng / ĩl

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)
Related products

collection of antibodies involved in photosynthesis

Plant protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution

1 : 25 000 with standard ECL (WB)

Expected | apparent MW

102 | 95 kDa

Confirmed reactivity

Arabidopsis thaliana, Zea mays

Predicted reactivity Hordeum vulagre
Not reactive in

no confirmed exceptions from predicted reactivity are currently known

Additional information

PPDK levels inr C3 plants like Arabidopsis thaliana and Hordeum vulgare are very low and PPDK protein is very dilute in most tissues of C3 plants. To perform detection in C3 plants leaf proteins needs to be concentrated before western blot, Chastain et al. (2002). 

Selected references to be added when available

Application example

western blot using anti-PPDK antibodies

5 µg of total protein from samples such as Zea mays leaf, were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved 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: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602, Agrisera) diluted to 1:50 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 5 minutes.

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