RbcL II | Rubisco large subunit, form II

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AS15 2955 |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity:  photosynthetic bacteria, archea, dinoflagellates


14 st
Item No:
AS15 2955

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product information

Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) catalyzes the rate-limiting step of CO2 fixation in photosynthetic organisms.

Form II Rubisco is present in many photosynthetic bacteria and archaea and in some photosynthetic dinoflagellates.

The large subunit (LSU) of form I Rubisco is encoded by the RbcL/cbbL gene, while form II LSU is encoded by cbbM.


KLH-conjugated synthetic peptide conserved in known RbcL form II protein sequences

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water.
Storage 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.
Tested applications Western blot (WB)
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AS07 259
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AS07 222
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matching secondary antibody

Plant and algal protein extraction buffer

Secondary antibodies

Additional information The antibody does not detect Form I RbcL/cbbL.
application information
Recommended dilution

1:10 000 (WB) 

Expected | apparent MW

51.7 kDa

Confirmed reactivity Amphidinium carterae, Rhodobacter capsulatus
Predicted reactivity Acidithiobacillus ferrooxidans, Dechloromonas aromatica, Gonyaulax polyedra, Leptothrix cholodnii, Magnetovibrio blakemorei, Rhodobacter sphaeroides, Rhodospirillum rubrum, Thiobacillus denitrificans, Rhodopseudomonas palustris

and with a signle mismatch in one amino acid:
Gallionella capsiferriformans, Mariprofundus ferrooxydans, Thioalkalicoccus limnaeus, Methanomethylovorans hollandica, Methanococcoides burtonii, Methanosaeta concili, Methanolobus tindarius, Methanohalophilus mahii, and the dinoflagellate Symbiodinium sp. (ex Stylophora pistillata)
Not reactive in Burkholderi
Additional information Protein extraction from diatoms protocol can be found here.
Selected references To be added when available, antibody released in September 2015

application example

western blot using anti-RbcL form II antibodies

1.5 µg of total protein extract from Rhodobacter capsulatus (1); extracted with Protein Extraction Buffer PEB (AS08 300); 0.5 pmol of recombinant RbcL I (2), 0.5 pmol of recombinant RbcL II (3) 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.

Western blot using RbcL II antibody on dinoflagelates
Quantitative Immunoblot of Form II RbcL from Amphidinium carterae ( Dinoflagellata ). Algae were extracted from PC filters in protein extrac%on buffer (AS08 300) with 3 breakage cycles (60 seconds, 6.5 m/s) in a FastPrep-24 with D-matrix ceramic beads (MP Biomedicals). Total algal protein extract (2 μg, lanes 1-8) and a range of loads of recombinant form II RbcL standard (AS15 2955S, lanes 9-13, 37.5, 75, 150, 300, 450 fmol) was separated on a Bolt polyacrylamide gel (ThermoFisher) and bloSed onto PVDF. Following antibody incubations (primary antibody AS15 2955, 1:20000; secondary antibody AS09 602, 1:20 000), protein signal was developed using ECL Select (Life Technologies) and detected using a VersaDoc Imager (BioRad).

Courtesy of Environmental Proteomics N.B

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