TKL1 | transketolase (chloroplastic)
AS15 2903 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana, Pisum sativum
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Approximately 1 ug (in dilution series) of total soluble (stromal) protein of Arabidopsis thaliana extracted from isolated chloroplasts using a buffer containing 10% glycerol, 200 mM NaCl, protease inhibitor and 1mM DTT. Samples were denatured at 96°C for 3 min and were separated on 10 % SDS-PAGE and blotted for 45min. to PVDF using semi-dry transfer. Blots were blocked with TBST with 5 % milk powder (0.02 % Tween 20) for 1h at room temperature (RT). Blot was incubated in the primary antibody at a dilution of 1: 5 000 for overnight at 4°C with agitation in TBST with 5 % milk powder. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 12:0 000 in for 1h at RT with agitation. The blot was washed as above and developed following manufacture recommendations. Exposure time was 10-30 seconds.
Courtesy of Dr. Ute Vothknecht and Edoardo Cutolo, Munich University, Germany
Antibody works on whole leaf extracts and isolated chloroplasts and is also recognizing recombinant TKL protein.
This product contains 10% glycerol and might appear as liquid but is provided lyophilized.