GlnA | glutamine synthetase positive control/quantitation standard

Glutamine synthetase (GlnA) is the key enzyme in the incorporation of mineral nitrogen into glutamine.This product is a recombinant GlnA protein standard, source Synechocystis strain PCC 6803.

Standard curve: 3 loads are recommended (0.5, 2 and 4 μl) .For most applications a sample load of 0.2 μg of chlorophyll will give a GlnA signal in this range.Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. 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.

Concentration:after adding 225  µl of milliQ water final concentration of the standard is 0.20 pmol/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), 50mM DTT.This standard is ready-to-load and does not require any additions or heating. It needs to be fully thawed and thoroughly mixed prior to using. Avoid vigorous vortexing, as buffers contain detergent. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.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 GlnA protein standard can be used in combination with global anti-GlnA antibodies to quantitate GlnA from a wide range of species. Global antibodies are raised against highly conserved amino acid sequences in the GlnA protein.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.