SHMT | Serine hydroxymethyltransferase
AS05 075 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, Citrus sp., P. sativum, S. oleracea, V. faba | cellular[compartment marker] of mitochondrial matrix
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Purified SHMT protein from Spinacia oleracea
53 kDa (Arabidopsis thaliana)
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30 µg of total protein from cauliflower Brassica oleracea var. botrytis mitochondrial fraction, extracted according to Rurek el al. 2015 and denatured with standard 1 x conc. Sample Buffer with 2-mercaptoethanol at 80°C for 10 min were separated on 12 % SDS-PAGE and blotted 1h to Immobilon-P using semi-dry transfer in the standard transfer buffer (20 % methanol, 48 mM Tris, 39 mM glycine, 0.0375 % SDS). Blots were blocked with 5 % skimmed milk in PBS-T with 0.01 % Tween20 for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody in 2 % skimmed milk in PBS-T in 2 % skimmed milk at a dilution of 1: 10 000 for 1h at RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera ) diluted to 1:10 000 in PBS-T with 2 % milk, for 1h at RT with agitation. The blot was washed as above and developed for min with enhanced chemiluminescence. Exposure time was 5 s.
Courtesy of Michal Rurek, D.Sc., Adam Mickiewicz University, Poznań (Poland)
This antibody can be used on total leaf extracts and isolated mitochondria
Serine hydroxymethyltransferase (SHMT) is part of the mitochondrial enzyme complex.
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