TKL1 | transketolase (chloroplastic)
AS15 2903 | clonality: polyclonal | host: rabbit | reactivity:Arabidopsis thaliana, Pisum sativum
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|Recommended dilution||1 : 2000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana, Pisum sativum
|Predicted reactivity||Synechococcus elongatus sp PCC 7942|
|Not reactive in|
|Additional information||TKL1 has MW of 79.28 kDa with and 73.45 kDa without presequence; about 72 kDa on SDS gel
Antibody works on whole leaf extracts and isolated chloroplasts and is also recognizing recombinant TKL protein.
|Selected references||Rocha et al. (2014). Phosphorylation of Arabidopsis transketolase at Ser428 provides a potential paradigm for the metabolic control of chloroplast carbon metabolism. Biochem J. 2014 Mar 1;458(2):313-22. doi: 10.1042/BJ20130631.|
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 (SERVALight EOS CL HRP WB Substrate Kit). Exposure time was 10-30 seconds.
Courtesy of Dr. Ute Vothknecht and Edoardo Cutolo, Munich University, Germany
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