RbcL | Rubisco large subunit, form I and form II (affinity purified)
AS03 037A | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, lichens, algae, cyanobacteria, dinoflagellates, diatoms | cellular [compartment marker] of plastid stroma in higher plants and cytoplasm in cyanobacteria
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KLH-conjugated synthetic peptide conserved across all known plant, algal and (cyano)bacterial RbcL protein sequences (form I L8S8 and form II L2), including Arabidopsis thaliana AtCg00490, Hordeum vulgare P05698, Oryza sativa P0C510, Chlamydomonas reinhardtii P00877, Synechococcus PCC 7920 A5CKC5
52.7 kDa (Arabidopsis thaliana), 52.5 kDa (cyanobacteria), 52.3 (Chlamydomonas reinhardtii)
Total protein from Populus T89 were extracted with “KEB buffer”, precipitated with ethanol on ice and denatured with “loading buffer” at 100°C for 10 min, separated on 8% SDS-PAGE and blotted O/N to PVDF using (wet blot) tank transfer. Blots were blocked with 5%TBS milk, for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 TBS for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly with TBS-T, then washed for 1h in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG HRP-conjugated, from Agrisera, AS09 602) diluted to 1:5000 in TBS-M (milk 5%) for 1h at RT with agitation. The blot was washed as above and developed with chemiluminescent detection reagent, for 10s increment until exposure time of 30s total.
Courtesy Dr. Mark Ruhl, Umeå Plant Science Centre, Sweden
This antibody is especially suitable for quantifying of Rubisco in plant and algal samples.
Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) catalyzes the rate-limiting step of CO2 fixation in photosynthetic organisms. It is demonstrably homologous from purple bacteria to flowering plants and consists of two protein subunits, each present in 8 copies. In plants and green algae, the large subunit (~55 kDa) is coded by the chloroplast rbcL gene, and the small subunit (15 kDa) is coded by a family of nuclear rbcS genes.
Ye et al. (2017). EMB2738, which encodes a putative plastid-targeted GTP-binding protein, is essential for embryogenesis and chloroplast development in higher plants. Physiol Plant. 2017 Jul 4. doi: 10.1111/ppl.12603.
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