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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Product Information
Immunogen
Purified SHMT protein from Spinacia oleracea
Host
Rabbit
Clonality
Polyclonal
Purity
Total IgG
Format
Lyophilized in PBS pH 7.4
Quantity
0.5 mg
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)
Recommended dilution
1: 1000 to 1 : 5 000 (WB)
Expected | apparent MW
53 kDa (Arabidopsis thaliana)
Reactivity
Confirmed reactivity
Arabidopsis thaliana, Brassica oleracea, Citrus sp., Oryza sativa, Pisum sativum, Spinacia oleracea, Vicia faba
Predicted reactivity
Chlamydomonas reinhardii, Citrus unshiu, Glycine max, Hordeum vulgare, Medicago truncatula, Populus balsamifera, Ricinus communis, Solanum tuberosum, Vitis vinifera
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known.
Application examples
Application examples
Application example
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)
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)
Additional information
This antibody can be used on total leaf extracts and isolated mitochondria
Background
Background
Serine hydroxymethyltransferase (SHMT) is part of the mitochondrial enzyme complex.
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