product information Background 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. Immunogen KLH-conjugated synthetic peptide derived from known CPN60 sequences, including Arabidopsis thaliana UniProt P21238 . TAIRAT2G28000.The peptide is conserved in chloroplastic CPN60A1 but NOT the close relative CPN60A2. Host Rabbit Clonality Polyclonal Clone Purity Serum Format Lyophilized Quantity 50 µl Reconstitution For reconstitution add 50 µl 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) Related products collection of antibodies to RUBISCOcollection of antibodies to proteins involved in photosynthesis Plant protein extraction buffer Secondary antibodies Additional information The antibody will work on loads from 5 µg/well.

application information
Recommended dilution		1 : 1000 with standard ECL (WB)
Expected | apparent MW		57.1 kDa (mature protein)
Confirmed reactivity		Arabidopsis thaliana, Arabidopsis thaliana cell culture, Nicotiana tabacum, Phaseolus vulgaris, Pisum sativm, Zea mays
Predicted reactivity		Aegilops squarrosa, Avicena marina, Brassica napus, Canavalia lineata, Narcissus pseudonarcissus, Oryza sativa, Ricinus communis, Trifolium pratense, Triticum aestivum
Not reactive in		cyanobacteria, algae
Additional information
Selected references		to be added when available, antibody released in September 2013.

application exampleApproximately 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

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