RbcL antibody and protein standard: Please store at -20 °C (6 months) or -80°Cfor long term storage(years). Please avoid freezing and thawing of reconstituted antibodies, make aliquots instead. PEB extraction buffer:Stable at RT for at least 1 month. For short-term storage please stoore (1 month) at 4°Cand for long term storage (1 year) at -20 °C.
Alpha proteobacteria, Algae (brown and red), Beta-proteobacteria, Dicots, Conifers, Cryptomonad, Cyanobacteria, Gamma-proeobacteria, Liverworts, Mosses, Prochlorophytes, PelwitschiaSpecies of your interest not listed? Contact us
Immunogen:
KLH-conjugated synthetic peptide was used to elicit anti-RbcL antibody. No baculovirus was used in production of this product.
1 x 50 µl of AS03 037, RbcL | Rubisco large subunit, form I and form II (amount enough for 50-100 Western blots)1 x 100 µl of AS01 017S, Rubisco protein standard (0.15 pmoles / µl, amount enough for generation of standard curve in 34 assays (standard curve: 0.0625 pmoles, 0.125 pmoles, 0.25 pmoles)2 x 2 ml of AS08 300, Protein extraction buffer (amount enough for 48 isolations of plant material, using 500 µl 1x PEB for 100 mg fresh weight) or 120 isolations of algal material (using 200 µl 1x PEB for cell amounts corresponding to 4-10 µg total chlorophyll)2 x 10 µl of AS09 602, Goat anti-rabbit IgG (H&L), HRP conjugated(amount enough for 50-100 Western blots)1 X 10 ml of AS16 ECL-N-10, AgriseraECL Bright 10 ml (5 ml of component A + 5 ml of component B, trial pack)
Quantitative western blot: detailed method description, video tutorialDiscussion over some critical aspects of quantitative western blot: Heidebrecht et al. (2009). Improved semiquantitative Western blot technique with increased quantification range. J. Immunological Methods. 35:40-48.
Defez et al. (2019). Bacterial IAA-Delivery into Medicago Root Nodules Triggers a Balanced Stimulation of C and N Metabolism Leading to a Biomass Increase. Microorganisms. 2019 Sep 29;7(10). pii: E403. doi: 10.3390/microorganisms7100403.Sorrentino et al. (2018). Performance of three cardoon cultivars in an industrial heavy metal-contaminated soil: Effects on morphology, cytology and photosynthesis. J Hazard Mater. 2018 Jun 5;351:131-137. doi: 10.1016/j.jhazmat.2018.02.044.Mota et al. (2015). Effects of heavy metals on Cyanothece sp. CCY 0110 growth, extracellular polymeric substances (EPS) production, ultrastructure and protein profiles. J Proteomics. 2015 Apr 29;120:75-94. doi: 10.1016/j.jprot.2015.03.004.Thamatrakoln et al. (2013). Death-specific protein in a marine diatom regulates photosynthetic responses to iron and light availability. Proc Natl Acad Sci U S A. 2013 Dec 10;110(50):20123-8. doi: 10.1073/pnas.1304727110. Supplemental material describes western blot quantification method.
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.
Host Animal:
Rabbit, Secondary antibody: Goat,
Species Reactivity:
Algae, Cyanobacteria, Higher plants
Immunogen:
KLH-conjugated syntetic peptides for respective antibodies, see product info sheets
50 µl of respective antibody, 100 µl of each protein standard, 2 x 10 µl of secondary antibody, 10 ml of ECL reagent,
Estimation of PSI to PSII ratio can be done using quantitative western blot technique using anti-PsaC (PSI) and PsbA (PSII) antibodies. References: Brown et al. (2007). Resource dynamics during infection of Micromonas pusilla by virus MpV-SP1. Environmental Microbiology 9(11): 2720-2727. Brown et al. (2008). Flux capacities and acclimation costs in Trichodesmium from the Gulf of Mexico. Marine Biology 154 (3): 413-422.
Selected references:
Abramson (2018). CARBON PARTITIONING IN ENGINEERED CYANOBACTERI UM FOR THE STUDY OF FEEDBACK INHIBITION OF PHOTOSYNTHESIS. Michigan State University, ProQuest Dissertations Publishing, 2018. 10826228.Morash et al. (2007) Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Can J. of Bot. 85: 476-483(8)Bouchard et al. (2006) UVB effects on the photosystem II-D1 protein of phytoplankton and natural phytoplankton communities. Photochem and Photobiol 82: 936-951.MacKenzie et al (2005). Large reallocations of carbon, nitrogen and photosynthetic reductant among phycobilisomes, photosystems and Rubisco during light acclimation in Synechococcus elongatus are constrained in cells under low environmental inorganic carbon. Arch of Microbiol. 183: 190 - 202.
Special application note:
Product information - Primary antibodies:Product number:Product name:Reconstitution: Recommended dilution:AS03 037Rabbit Anti-RbcL Global antibodyFor reconstitution see lable on respective tube.1:5000-10 000 with ECLAS10 939Rabbit Anti-PsaC Global antibodyFor reconstitution see lable on respective tube.1:1000 with ECLAS05 084Rabbit Anti-PsbAGlobal antibodyFor reconstitution see lable on respective tube.1:10 000 with ECL* All primary antibodies in this kit are raised in rabbits.Product information - Protein standards:Product number:Product name:Concentration:Size:Western Blot – Positive Control:AS01 016SPsbA *0.25 pmol/ μl41.5 kDa#To generate a standard curve, 3 loads are suggested (0.5, 2 and 4 ul). For most applications a sample load of 0.2 ug of chlorophyll will give a PsbA signal in this range.A 2 ul load is optimal for most chemiluminescent detection systems to use as a positive control.AS01 017SRbcL *0.15 pmol/ μl52.7 kDaTo generate a standard curve, 3 loads are suggested (0.5, 2 and 4 ul). For most applications a sample load of 0.2 ug of chlorophyll will give a RbcL signal in this range.A 2 ul load is optimal for most chemiluminescent detection systems to use as a positive control.AS04 042SPsaC *0.15 pmol/μl11.5 kDa#To generate a standard curve, 3 loads are suggested (0.5, 2 and 4 l). For most applications a sample load of 0.2 g of chlorophyll will give a PsaC signal in this range.A 2 ul load is optimal for most chemiluminescent detection systems as a positive control.*These proteins are larger than a respective native protein due to the addition of His-tag* For reconstitution of standards see lable on respective tube.Product information - Secondary antibody:AS09 602-trial Goat anti-Rabbit IgG (H&L), HRP conjugated, 20 l (2x10 l)Product information - ECLreagent:AS16 ECL-N-10 AgriseraECL Bright (10 ml trial pack)Educational information about Quantitative western blot can be found here: detailed method description, video tutorial
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.
Product Type:
Antibody
Format:
Lyophilized in glycerol.
Storage Temp:
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.
Concentration: after re-constitution with sterile milliQ water final concentration of the standard is 0.15 pmoles/ lProtein 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.
Application Details:
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 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 does not require heating before loading on the gel or addition of any buffer.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.
Reconstitution:
For reconstitution add 90 l of sterile water, Please notice that this product contains 10% glycerol and might appear as liquid but is provided lyophilized
Molecular Weight:
52.7 kDa
Selected references:
Capo-Bauca et al. (2023). Carbon assimilation in upper subtidal macroalgae is determined by an inverse correlation between Rubisco carboxylation efficiency and CO2 concentrating mechanism effectiveness. New Phytol. 2023;237(6):2027-2038. doi:10.1111/nph.18624Capo-Bauca et al. (2022) Correlative adaptation between Rubisco and CO2-concentrating mechanisms in seagrasses. Nat Plants. 2022 Jun;8(6):706-716. doi: 10.1038/s41477-022-01171-5. Epub 2022 Jun 20. Erratum in: Nat Plants. 2022 Jun 29;: PMID: 35729266.Perera-Castro et al (2022). Limitations to photosynthesis in bryophytes: certainties and uncertainties regarding methodology. J Exp Bot. 2022;73(13):4592-4604. doi:10.1093/jxb/erac189Poor et al. (2018). Comparison of changes in water status and photosynthetic parameters in wild type and abscisic acid-deficient sitiens mutant of tomato (Solanum lycopersicum cv. Rheinlands Ruhm) exposed to sublethal and lethal salt stress. J Plant Physiol. 2018 Dec 8;232:130-140. doi: 10.1016/j.jplph.2018.11.015.Dai et al. (2018). Visualizing Individual RuBisCO and its Assembly into Carboxysomes in Marine Cyanobacteria by Cryo-Electron Tomography. J Mol Biol. 2018 Aug 20. pii: S0022-2836(18)30411-X. doi: 10.1016/j.jmb.2018.08.013.
Special application note:
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
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.
Product Type:
Antibody
Antibody Type:
Polyclonal
Format:
Liquid
Storage Temp:
Store at 4 C; make aliquots to avoid working with a stock. 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.
KLH-conjugated synthetic peptide derived from all known plant,algal and cyanobacterial RbcL (Rubisco large subunit of Rubisco Form I) sequences, including Arabidopsis thaliana UniProt: O03042, TAIR: AtCg00490, Synechococcus sp. Q3ALL1
Applications:
Immunogluorescence (IF), Immunolocalization (IL), ImmunoGold (IG), Western blot (WB)
This antibody detects RbcL protein from 102.6 fmoles and has been used as a control to ensure adequate permeabilization and fixation of toxic cyanobacterial cells in immunolabeling experiments (method based on: Orellana & Perry (1995) J Phycol 31: 785-794).Antibody has been used in immunolabelling of intact cyanobacterial cells fixed with ethanol using a secondary anti-IgY antibody conjugated with a fluorochrome.For Rubisco quantification using quantitative western blot technique, anti-RbcL antibody, (AS03 037) combined with Rubisco ready to use standard (AS01 017) is recommended.
Application Details:
(IL) tested on a grass species, formaldehyde-fixed and paraffin-embedded tissue following the protocol from Gonzalez et al, (1998) Plant Physiol, V, 116, 1 : 10 000-1 : 20 000, 2 g of total cellular protein, (WB)
Purity:
Purified, total IgY (chicken egg yolk immunoglobulin) in PBS pH 8. Contains 0.02 % sodium azide.
No confirmed exceptions from predicted reactivity are currently known
Selected references:
Guljamow et al. (2021) Diel Variations of Extracellular Microcystin Influence the Subcellular Dynamics of RubisCO in Microcystis aeruginosa PCC 7806. Microorganisms. 2021 Jun 10;9(6):1265. doi: 10.3390/microorganisms9061265. PMID: 34200971; PMCID: PMC8230624. (IF)Morin et al. (2019). Morin et al. (2019). Response of the sea-ice diatom Fragilariopsis cylindrus to simulated polar night darkness and return to light. Limnology and Oceanography. 9999, 2019, 1–20. (sea-ice diatom)Lv et al. (2019). Uncoupled Expression of Nuclear and Plastid Photosynthesis-Associated Genes Contributes to Cell Death in a Lesion Mimic Mutant. Plant Cell. 2019 Jan;31(1):210-230. doi: 10.1105/tpc.18.00813.Gell rt et al. (2018). A single point mutation on the cucumber mosaic virus surface induces an unexpected and strong interaction with the F1 complex of the ATP synthase in Nicotiana clevelandii plants. Virus Res. 2018 Jun 2;251:47-55. doi: 10.1016/j.virusres.2018.05.005.Robert et al. (2015). Leaf proteome rebalancing in Nicotiana benthamiana for upstream enrichment of a transiently expressed recombinant protein. Plant Biotechnol J. 2015 Aug 19. doi: 10.1111/pbi.12452.
Special application note:
Peptide target used to elicit this antibody is not conserved in type II Rubisco found in dinoflagellates and some photosynthetic bacteria
UniProt number:
O03042 , Q3ALL1
TAIR number:
ATCG00490
Research area:
Global Antibodies
Code:
Photo20 CM20
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