RbcL | Rubisco large subunit, form I

PRODUCT INFORMATION IN PDF

product information 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. Immunogen 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 Host Chicken Clonality Polyclonal Clone Purity Total IgY in PBS pH 8.0+ 0.02% sodium azide Format Liquid Quantity 50 µl (16 mg/ml) Reconstitution Storage Store at 4°C; make aliquots to avoid working with a stock. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from liquid material adhering to the cap or sides of the tubes. Tested applications Immunolocalization (IL), Western blot (WB) Related products 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) AS15 2955 | anti-RbcL II | Rubisco large subunit, form II (50 µl), rabbit antibody AS15 2955S | RbcL II | Rubisco form II positive control/quantitation standard AS01 017S | Rubisco protein standard for quantitative western blot or positive control AS03 037PRE | Rubisco large subunit, pre-immune serum AS09 409 | Rubisco quantitation kit AS15 2994 | Rubisco ELISA quantitation kit  AS07 218  | anti-Rubisco | 557 kDa hexadecamer, rabbit antibody to a whole protein AS07 259 | anti-RbcS | Rubisco small subunit (SSU), rabbit antibody AS07 222 | anti-RbcS | Rubisco small subunit (SSU) from pea, rabbit antibody Recommended secondary antibodies: Goat anti-chicken IgY (H&L), HRPconjugated or Goat anti-chicken IgY (H&L), ALP conjugated Plant and algal protein extraction buffer Secondary antibodies Additional information Peptide target used to elicit this antibody is not conserved in type II Rubisco found in dinoflagellates and some photosynthetic bacteria. For those species using product AS03 037 is recommended.

application information
Recommended dilution		(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 µg of total cellular protein, (WB)
Expected | apparent MW		52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 kDa (Chlamydomonas reinhardtii)
Confirmed reactivity		Arabidopsis thaliana, Lobaria pulmonaria, Medicago sativa, mixed phytoplankton, Pinus strobus, Pisum sativum, Solanum tuberosum, Spartina alterniflora, Spinacia oleracea, Synechococcus sp. PCC7842, Thiobacillus sp. Ulmus sp.
Predicted reactivity		Algae, Dicots, Conifers, Liverworts, Mosses, Prochlorophytes, Welwitschia
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
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.
Selected references		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.

application example



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, recommended 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 TMA-6 (Lumigen) 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.

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