PEPC | Phosphoenolpyruvate carboxylase positive control/quantitation standard

Phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) serves as an important control element in the regulation of photosynthetic carbon metabolism in C4 and CAM plants. This is the first enzyme of the pathway, and PEPC enzymes are encoded by a small multigenic family. Several isoforms of PEPC have been characterised in maize, sorghum and sugarcane. These isoforms are involved in several functions such as the initial fixation of atmospheric CO2 (= C4 PEPC) and anaplerotic functions associated with nitrogen assimilation or amino acid biosynthesis (Lepiniec et al. 1994) .

Standard curve: 3 loads are recommended (0.5, 2 and 4 μl) .For most applications a sample load of 0.2 μg of chlorophyll/well will give a RbcL signal in this range.Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. Higher standard concentration needs to be used to allow detection by Coomasie stains. Such gels with higher standard concentration can not be used for quantitation using chemiluminescence.This standard is stabilized and ready and does not require heating before loading on the gel. Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.

For reconstitution add 60 µl of steril water, Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized

Concentration:after re-constitution with sterile milliQ water final concentration of the standard is 0.15 pmoles/µLProtein standard buffer composition:Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51 mM, SERVA® Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1 mg/mL PefaBloc protease inhibitor (Roche), 50 mM DTT.This standard is ready-to-load and does not require any additions or heating.This standardis stabilized and readyand does not require heating before loading on the gel.Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.

The PEPC protein standard can be used in a combination with Agrisera global PEPC antibiody to quantitate PEPC from a wide range of species. Global antibodies are raised against highly conserved amino acid sequence. Quantitative western blot: detailed method description, video tutorial

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.