RbcL | Rubisco positive control/quantitation standard
Biozol Catalog Number:
AGR-AS01-017S
Supplier Catalog Number:
AS01-017S
Alternative Catalog Number:
AGR-AS01-017S
Manufacturer:
Agrisera
Category:
Proteine/Peptide
Application:
WB
Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) catalyzes the rate-limiting step of CO2 fixation in photosynthesis. It is one of the most abundant proteins on Earth and its homology has been demonstrated from purple bacteria to flowering plants
Molecular Weight:
52.7 kDa
Form:
Lyophilized in glycerol.
Antibody Type:
Secondary Antibody
Application Dilute:
Standard curve: three protein standard loads are recommended.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 syst
Application Notes:
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, 0.1mg/ml PefaBloc protease inhibitor (Roche), 50 mM 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 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.Please, use the 55 kDa size of RbcL for calculations. The pmoles in the standard refer to pmoles of rbcL monomers.Why can I not see the standard band using Coomasie stain? The reason that you do not see Rubisco standard on a gel is, that you have probably used it in concentration which is recommended for western blot detection, and it is too low to allow to see this protein using Coomasie stain. In such a case, you should load more Rubisco standard on a gel and stain it with more sensitive Coomasie stain or with silver. You can not use such a gel for western blot, as using higher concentration of this standard will not work for quantitation using western blot technique.
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