AtpB | Positive control/quantitation standard
Product Specifications
Background
ATP synthase is the universal enzyme that stnthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient.This product is a recombinant protein standard, source: Synechocystis strain PCC 6803.
CAS Number
9007-83-4
HS Code
38221900
Applications
Western blot (WB)
Field of Research
Photosynthesis protein standards-quantitation
Dilution
Standard curve: 3 loads are recommended (0.5, 2 and 4μl) .For most applications a sample load of 0.2μg of chlorophyll will give a AtpB signal in this range.Positive control: load per well: a 2μl load 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.
Format
Lyophilized, in glycerol.
Reconstitution
For reconstitution add 90 µl of milliQ water, Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized
Molecular Weight
In most gel systems AtpB migrates around 50-54 kDa
Precautions
Concentration:after adding 90 µl of dest. water final concentration of the standard is 0.27 pmol/µL.Protein 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, 0.1mg/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.
Additionnal Information
The AtpB protein standard can be used in combination with global anti-AtpB antibodies to quantitate AtpB from a wide range of species. Global antibodies are raised against highly conserved amino acid sequences in the AtpB protein.Quantitative western blot: detailed method description, video tutorial
References & Citations
Fraser et al. (2013) . Photophysiological and Photosynthetic Complex Changes during Iron Starvation in Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. PLOS ONE.
Storage Conditions
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.
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