RbcL | Rubisco positive control/quantitation standard

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Aplikacje: western blot (WB)
Zalecane rozcieńczenie: Standard curve: 3 loads are recommended (0.5, 2 and 4μl).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.
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Przechowywanie: 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.
Uwagi: The RbcL protein standard can be used in a combination with Agrisera global antibiodies (AS01 017 from chicken or AS03 037 from a rabbit) to quantitate RbcL from a wide range of species. Global antibodies are raised against highly conserved amino acid sequence. This standard is also included in following kits: Educational antibody kit - photosynthesis, Photosynthesis Tool Kit - quantitation, Rubico quantitation kit,Quantitative western blot: detailed method description, video tutorial
Powiązane produkty: Primary antibodies matching Rubisco standard are: AS03 037 | anti-RbcL | Rubisco large subunit, form I and form II (50 µl)AS03 037A | anti-RbcL | Rubisco large subunit, form I and form II (50 µg affinity purified)AS03 037-HRP| anti-RbcL | Rubisco large subunit, form I and form II (40 µg, HRP-conjugated) AS01 017 | anti-RbcL | Rubisco large subunit, form I, chicken antibodyAS15 2994 | Rubisco ELISA quantitation kit matching secondary antibodycollection of other protein standardscollection of other global antibodies
Dodatkowe informacje: 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.Please, use the 55 kDa size of RbcL for calculations. The pmoles in the standard refer to pmoles of rbcL monomers.

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.Source of Rubisco standard: Rubisco protein was purified directly from a plant tissue - spinach.
Literatura: Young et al. (2015). Antarctic phytoplankton down-regulate their carbon-concentrating mechanisms under high CO2 with no change in growth rates. Marine Ecology Progress Series 532:13-28. Vandenhecke et al. (2015). Changes in the Rubisco to photosystem ratio dominates photoacclimation across phytoplankton taxa. Photosynth Res. 2015 Apr 11. Wu et al. (2014). Large centric diatoms allocate more cellular nitrogen to photosynthesis to counter slower RUBISCO turnover rates. Front. Mar. Sci., 09 December 2014 | doi: 10.3389/fmars.2014.00068. Li et al. (2014). The nitrogen costs of photosynthesis in a diatom under current and future pCO2. New Phytol. 2014 Sep 25. doi: 10.1111/nph.13037.