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Anti-SHMT1 | Serine hydroxymethyltransferase 1, mitochondrial
Product no: AS23 4913
AS23 4913 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brassica napus
319
€
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- Product Info
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Immunogen: KLH-conjugated unique peptide derived from Arabidopsis thaliana SHMT1 protein sequence. UniProt: Q9SZJ5 TAIR: AT4G37930 The peptide is not conserved in the sequence of AtSHMT2.
Host: Rabbit Clonality: Polyclonal Purity: Antigen affinity purified serum, in PBS pH 7.4 Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution add 50 µl, of sterile or deionized 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 (WB) Expected | apparent MW: 57.4 | 54 kDa (due to N-terminal or processing) - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Brassica oleracea, Brassica rapa, Camelina sativa, Capsella rubella,Cardamine amara subsp. amara,Tarenaya hassleriana, Raphanus sativus, Vitis vinifera
Species of your interest not listed? Contact usNot reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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20 µg of crude mitochondrial extract protein from Arabidopsis thaliana leaves [1], (2) 10 µg of total soluble proteins from isolated mitochondria from Solanum tuberosum [2], (3) 10 µg of total soluble proteins from isolated mitochondria from Arabidopsis thaliana leaves, and (4) 6 µg of IMTACT isolated mesophyll mitochondrial proteins from Arabidopsis thaliana [3-4]. Proteins were separated on a reducing 12% polyacrylamide gel and blotted 1h to nitrocellulose. Membranes were blocked immediately following transfer in 5 % milk in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with gentle shaking. Membranes were incubated with the primary antibody at a dilution of 1:4000 overnight at 4°C with gentle shaking. The primary antibody solution was discarded, and the membrane was washed 3 times for 5 min in TBS-T at room temperature. The membrane was then incubated in secondary antibody (anti-rabbit IgG goat radish peroxidase conjugated, AS09 602) diluted to 1:10000 in TBS-T for 1h at room temperature with gentle shaking. The membrane was washed 3 times for 5 min in TBS-T at room temperature with gentle shaking and revealed for 5 min with chemiluminescent detection reagent according to the manufacturer’s instructions (Agrisera ECL kit bright; AS16 ECL-N-100). Images of the blots were obtained using an Azure c600 Western Blot Imaging system (Azure biosystems). Exposure time was 45 seconds (standard sensitivity).
Courtesy of Drs Boussardon and Keech, Umeå Plant Science Centre, Sweden
References:
[1] Keech O, Dizengremel P, Gardeström P (2005). Preparation of leaf mitochondria from Arabidopsis thaliana. Physiologia Plantarum 124: 403-409.
[2] Sani MA, Keech O, Gardeström P, Dufourc EJ and Gröbner G (2009). Magic Angle Phosphorus NMR of functional Mitochondria: in situ Monitoring of Lipid Response under Apoptotic-like Stress. FASEB Journal 23: 2872-2878
[3] Boussardon C, Przybyla-Toscano J, Carrie C and Keech O (2020). Tissue-Specific Isolation of Arabidopsis/plant Mitochondria - IMTACT (Isolation of Mitochondria TAgged in specific Cell Types). The Plant Journal 103:459-473
[4] Boussardon C and Keech O (2022). "Cell Type-Specific Isolation of Mitochondria in Arabidopsis." Methods Mol Biol 2363: 13-23 - Background
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Background: SHMT1 is an enzyme of the photorespiratory pathway, which is catalyzing the interconversion of serine and glycine. Involved in controlling cell damage caused by abiotic stress, such as high light and salt and the hypersensitive defense response of plants. - Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts
Agrisera Educational Poster Collection - Reviews:
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