CPN60A1 | Chaperonin 60 subunit alpha 1 (chloroplastic)
AS12 2613 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. arietinum, N. tabacum, P. vulgaris, P. sativum, Z. mays
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KLH-conjugated synthetic peptide derived from known CPN60 sequences, including Arabidopsis thaliana UniProt P21238 . TAIR AT2G28000.The peptide is conserved in chloroplastic CPN60A1 but NOT the close relative CPN60A2.
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Approximately 50-70 µg of total chloroplast or cell protein was extracted from various species by boiling in 4x Sample buffer for 5 min. These proteins were separated on 15 % Tris-Glycine SDS-PAGE run at constant voltage of 100V for 20min and then run at constant current of 15 mA for 1 hrs. Following separation, the proteins were transferred by electroblotting to PVDF (1h 30 min) using 1X Transfer buffer (14.4 gm glycine, 3 gm Tris-base, 200 ml MeOH in 1l ddH2O) pH 8.3. Blots were blocked with TBS with 1% Tween and 3% NFM (TBST w/NFM)) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for 1h at RT with agitation, and then left in 4°C overnight. The antibody solution was decanted and the blot was rinsed briefly three times, and then washed 3 times X 10 min in TBST w/NFM at RT with agitation. The blot was incubated in secondary antibody (Donkey anti-rabbit IgG HRP-conjugated) diluted to 1:15 000 in TBST for 1h at RT with agitation. The blot was washed as above and developed using Thermo SuperSignal West Pico Chemiluminescent Substrate reagent according to the manufacturer’s instructions and imaged on a Bio-Rad ChemiDoc Imager using an exposure time of 80 seconds.
Courtesy of Dr. Barry Bruce lab, University of Tennesee-Knoxville, USA
CPN60 alpha 1 (chaperonin 60 subunit alpha 1, chloroplastic) binds small and large subunits of Rubisco. Assists in protein folding and required for proper chloroplast development. Synonymes: protein SCHLEPPERLESS, RuBisCO large subunit-binding protein subunit alpha 1.
Lande et al. (2019). Dehydration-induced alterations in chloroplast proteome and reprogramming of cellular metabolism in developing chickpea delineate interrelated adaptive responses. Plant Physiology and Biochemistry Volume 146, January 2020, Pages 337-348.
Dogra et al. (2019). Impaired PSII proteostasis triggers an UPR-like response in the var2 mutant of Arabidopsis thaliana. J Exp Bot. 2019 Apr 16. pii: erz151. doi: 10.1093/jxb/erz151.
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