RbcL | Rubisco large subunit, form I (chicken)
AS01 017 | Clonality: Polyclonal | Host: Hen | Reactivity: [global antibody] for higher plants, algae, cyanobacteria | cellular [compartment marker] of plastid stroma
Benefits of using this antibody
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Product Information
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
52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 kDa (Chlamydomonas reinhardtii)
Reactivity
Application examples
1 µg of total protein from samples such as Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2), Zea mays leaf (3), Chlamydomonas reinhardtii total cell (4), were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, secondary antibody AS10 1489, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
Reactant: Euglena
Application: Western Blotting
Pudmed ID: 27391690
Journal: PLoS One
Figure Number: 2A
Published Date: 2016-07-09
First Author: Záhonová, K., Füssy, Z., et al.
Impact Factor: 2.942
Open PublicationAbundance of the RBCS and RBCL proteins in Euglena gracilis and Euglena longa.Protein immunodetection was performed using anti-RBCS, anti-RBCL, and anti-Tubulin antibodies. Three bands with different molecular weights were observed in anti-RBCS immunoblotting. The ~130 kDa band (marked *1) corresponds to polyprotein synthesized in the nucleus. The ~15 kDa band (marked *3) corresponds to the processed monomer after cleavage of the signal sequence and excision of decapeptides. The ~22 kDa band (marked *2) possibly corresponds to a monomer still attached to the transit peptide. The identity of the RBCL protein (arrowhead in the anti-RBCL panel) was confirmed by mass-spectrometry. Tubulin served as a loading control. Molecular weights in kDa are indicated on the left. EG-, E. gracilis cultivated photosynthetically (without ethanol); EG+, E. gracilis cultivated mixotrophically (with ethanol); EL, E. longa.
Reactant: Euglena
Application: Western Blotting
Pudmed ID: 27391690
Journal: PLoS One
Figure Number: 4A,B,C
Published Date: 2016-07-09
First Author: Záhonová, K., Füssy, Z., et al.
Impact Factor: 2.942
Open PublicationStability of RBCS and RBCL proteins in Euglena gracilis and Euglena longa.Cell cultures were treated with 20 ?g/ml of cycloheximide, aliquots were taken at 0, 1, 4, 8, and 24 h post treatment, and analyzed by western blotting using anti-RBCS, anti-RBCL and anti-Tubulin antibodies. Molecular weights (in kDa) are indicated on the left of each panel. The identity of the RBCL protein (arrowhead in the anti-RBCL panel) was confirmed by mass-spectrometry. Tubulin served as a loading control. Cultivation conditions and species are denoted as in Fig 2.
Additional information
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.
Background
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 citations
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.
Gellért 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.
Morash et al. (2007). Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Canadian J. of Botany, 2007, 85(5): 476-483, 10.1139/B07-043.
MacKenzie et al. (2005). Inorganic carbon acclimation in Synechococcus elongatus alters the dynamics of macromolecular pooks and photosynthetic fluxes in response to increased light. Photosynt Research 85: 341-357.
Schofield et al. (2003). Changes in macromolecular allocation in nondividins algal symbionts allow for photosynthetic acclimation in the lichen Lobaria pulmonaria. New Phytol 159: 709-718.
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