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.
Rurek et al. (2018). Mitochondrial Biogenesis in Diverse Cauliflower Cultivars under Mild and Severe Drought. Impaired Coordination of Selected Transcript and Proteomic Responses, and Regulation of Various Multifunctional Proteins, Int. J. Mol. Sci. 2018, 19(4), 1130; doi:10.3390/ijms19041130 (Special issue: Plant Mitochondria)
Rurek et al. (2018). Cold and Heat Stress Diversely Alter Both Cauliflower Respiration and Distinct Mitochondrial Proteins Including OXPHOS Components and Matrix Enzymes, Int. J. Mol. Sci. 2018, 19(3), 877; doi:10.3390/ijms19030877 (Special issue: Plant Mitochondria)
Yin et al. (2016). Comprehensive Mitochondrial Metabolic Shift during the Critical Node of Seed Ageing in Rice. PLoS One. 2016 Apr 28;11(4):e0148013. doi: 10.1371/journal.pone.0148013. eCollection 2016.
Long et al. (2015). Contributions of photosynthetic and non-photosynthetic cell types to leaf respiration in Vicia faba L. and their responses to growth temperature. Plant Cell Environ. 2015 Apr 1. doi: 10.1111/pce.12544.
Wei et al. (2013). Folate polyglutamylation eliminates dependence of activity on enzyme concentration in mitochondrial serine hydroxymethyltransferases from Arabidopsis thaliana. Arch Biochem Biophys 1;536(1):87-96.
Camejo et al. (2012).Salinity-inducedchanges inS-nitrosylation ofpeamitochondrialproteins. J. Proteomics Dec.11.