product information

background
 

PEPC (phosphoenolpyruvate carboxylase), EC=4.1.1.31, belongs to an enzyme family of carboxy-lyases that is catalyzing adding fo carbon dioxide to phosphoenolpyruvate (PEP) to form oxaloacetate.  Alternative names: PEPCase 1, PEPCase 3, PEPC 1, PEPC 3

immunogen
 

KLH-conjugated synthetic peptide well conserved PEPC1 and sequences from different plant species including Arabidopsis thaliana Q9MAH0  (PEPC 1), Q84VW9 (PEPC 3). The peptide chosen to elicit this antibody is also perfectly conserved in bacterial type of this enzyme NP_177043.2 (PEPC 4)

antibody format
 

rabbit

polyclonal

affinity purified serum in PBS pH 7.4,

lyophilized

quantity
 

200 µg

for reconstitution add 200 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)

related products
 

AS07 241 | anti-PEPCK |  PEP carboxy kinase

additional information
 

to be added when available

application information

recommended dilution
 

1 : 1000 with standard ECL (WB)

expected | apparent MW
 

110 | 105 kDa

confirmed reactivity
 

Arabidopsis thaliana, Hordeum vulgare, Zea mays, Phaeodactylum tricornutum (strain CCAP 1055/1

predicted reactivity
 

dicots including: Cucumis sativus, Lupinus albus, Solanum tuberosum, Spinacia oleracea, monocots including: Triticum aestivum, Oryza sativa, algae, diatoms: Thalassiosira pseudonana

other species: Salmonella sp. Shigella sp. Yersinia sp. Vibrio sp.

not reactive in
 

no confirmed exceptions from predicted reactivity known in the moment

additional information
 

to be added when available

selected references
 

to be added when available, antibody released in August 2009

application example

5 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Protein Extration Buffer, PEB (AS08 300), (2) Spinacia oleracea  total cell, extracted with PEB, (3 )Hordeum vulgare total cell extracted with PEB, (4) Zea mays total cell extracted with PEB, 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: 10 000 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-rabbit IgG horse radish 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 manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).

 

 

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