The Rubisco quantitation kit is suitable for quantitation of Rubisco in plant and algal samples using quantitative immunoblotting. It contains anti-RbcL antibody, Rubisco protein standard and the Protein Extraction Buffer for quantitative protein extraction. This kit is a good alternative to quantification of Rubisco by ELISA as it allows separation of Rubisco from a total complex cell protein mixture, which can be troublesome in case of ELISA method.
Product Type:
Antibody
Antibody Type:
Polyclonal
Storage Temp:
RbcL antibody and protein standard: Please store at -20°C (6 month) or -80°C for 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°C and 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)5 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.
Application Details:
1 : 5000-1 : 10000 (WB)
Related products:
Rubisco/Carbon metabolism antibody collectionAS15 2994 | Rubisco ELISA quantitation kit AS15 2955 | Anti-RbcL II | Rubisco large subunit, form II (50 µl), rabbit antibodiesAS15 2955S | RbcL II | Rubisco form II positive control/quantitation standardAS01 017 | Anti-RbcL | Rubisco large subunit, form I, chicken antibodiesAS01 017S | Rubisco protein standard for quantitative western blot or positive controlAS03 037PRE | Rubisco large subunit, pre-immune serumAS07 218 | Anti-Rubisco | 557 kDa hexadecamer, rabbit antibodies, to a whole protein AS07 259 | Anti-RbcS | Rubisco small subunit (SSU), rabbit antibodiesmatching secondary antibody
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.
Agrisera
AS05 070
10 or 50 µl / tube of each antibody. 100 µl / tube of each protein standard. 50 µl (5 x 10 µl) of secondary antibody. 10 ml trial pack of ECL reagent.
This kit contains antibodies to the following 5 proteins and 4 protein standards for their quantification. It also includes a matching secondary antibody and an ECL detection reagent.Rubisco - universal and absolutely essential carbon fixation enzyme that is conserved across oxygenic photoautotrophs and other groups. The RbcL (Rubisco large subunit) protein is hyperabundant and regulation of RbcL content under different conditions or stress responses sets a limit on the maximum capacity for CO2 uptake in a population or community.Proteins involved in conversion of solar energy into chemical bonds:PsaC protein of multisubunit protein complex - Photosystem I, which harnesses light energy required for photosynthesis to occur.PsbA protein of multisubunit complex - Photosystem II - the ultimate source of almost all biosynthetic reductant in the biosphere. PsbA (D1) protein is rapidly cycled under illumination in all oxygenic photobionts. Tracking PsbA pools using the Global PsbA Antibody shows the functional content of Photosystem II in a wide range of samples.Lhcb1 - harnesses light energy for photosynthesis (Photosystem II)AtpB highly conserved across the beta subunits of known F-type ATP Synthases from chloroplasts, mitochondria and most bacteria. The ATP Synthase complex is essential for the synthesis of ATP from ADP and free phosphate
Product Type:
Antibody
Antibody Type:
Polyclonal
Storage Temp:
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.
Photosynthetic antibody collectionPlant and algal protein extraction buffer Secondary antibodies
Special application note:
This kit can be used during teaching labs and contains following antibodies and protein standards:Product information - Primary antibodies: Product number: Product name: Reconstitution: Recommended dilution: AS01 004Anti-Lhcb1 antibody*Please follow the instructions on the lable on each vial.1:2000 for ECL detection.AS03 037Anti-RbcL global antibody* Please follow the instructions on the lable on each vial.1:5000-10 000 for ECL detection.AS10 939Anti-PsaC global antibody*Please follow the instructions on the lable on each vial.1:1000 for ECL detection.AS05 084Anti-PsbA antibody*Please follow the instructions on the lable on each vial.1:10 000 for ECL detection.AS05 085-10Anti-AtpB antibody*Please follow the instructions on the lable on each vial.1:2000-5000 for ECL detection.* All primary antibodies in this kit are raised in rabbits.Product information - Protein standards/positive controls:Product number:Product name:Reconstitution:Amount to load on the gel:Size:AS01 016SPsbA protein standard Please follow the instructions on the lable on each vial.2 μl (0.25 pmol/ μl)41.5 kDa*AS01 017SRbcL (Rubisco) protein standardPlease follow the instructions on the lable on each vial.2 μl (0.056 pmol/ μl)52.7 kDaAS03 030SAtpB protein standardPlease follow the instructions on the lable on each vial.10 μl (0.27 pmol/μl)53.1 kDa*AS04 042SPsaC protein standard Please follow the instructions on the lable on each vial.2 μl (0.15 pmol/μl)11.5 kDa**These proteins are larger than a respective native protein due to the addition of His-tag Product information - Secondary antibodiy:AS09 602- Goat anti-Rabbit IgG (H&L), HRP conjugated, 50 µl (5x10 µl)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
This kit allows quantitation of major photosynthetic proteins: Rubisco, PsaC (PSI) and PsbA (PSII) as well as determination of PSI/PSII ratios using the western blot technique. The kit contains following global antibodies, protein standards and secondary antibody: Rabbit anti-RbcL (Form I). Product number: AS03 037 Rabbit anti-PsaC. Product number: AS10 939Rabbit anti-PsbA Product number: AS05 084Rubisco protein standard. Product number: AS01 017S PsaC protein standard. Product number: AS04 042S PsbA protein standard. Product number: AS01 016S Goat anti-Rabbit IgG (H&L), HRP conjugated (5 x 10 µl), Product number: AS09 602-trialAgriseraECL Bright (10 ml trial pack), Product number: AS16 ECL-N-10
Product Type:
Antibody
Antibody Type:
Polyclonal
Storage Temp:
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.
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. 5 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.
Related products:
Plant and algal protein extraction bufferSecondary antibodies
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 µl). For most applications a sample load of 0.2 µg 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, 50 µl (2x25 µl)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
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