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SUN1,2 | SUN domain-containing protein 1,2
AS18 4224 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
- Product Info
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Immunogen: KLH-conjugated peptide derived from N-terminus of SUN1 of Arabidopsis thaliana UniProt: Q9FF75 , TAIR: AT5G04990and SUN2, UniProt: Q9SG79, TAIR: AT3G10730 Host: Rabbit
Clonality: Polyclonal Purity: Immunogen affinity purified serum in PBS pH 7.4. Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution add 50 µl, of sterile water Storage: Lyophilized antibody can be stored at -20°C for up to 3 years. Re-constituted antibody can be stored at 4°C for several days to weeks. Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube. Tested applications: Immunofluorescence (IF), Western blot (WB) Recommended dilution: 1 : 1000 (WB) Expected | apparent MW: 51 | 56 kDa - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Arabis nemor, Eutrema salsugineum, Microthlaspi erraticum, Noccaea caerulescens Species of your interest not listed? Contact us
Not reactive in: Pisum sativum - Application Examples
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Immunofluorescent localization of SUN1,2 (red) on a male meiocyte from Arabidopsis thaliana. DAPI - grey. Detailed method can be found in Hurel et al. 2018.
Courtesy of Dr. Mathilde Crelon, INRAE, France
Samples:
1 - 15 ug of Arabidopsis thaliana Col-0 callus total protein
2 - 15 ug of Arabidopsis thaliana sun1 mutant callus total protein
3 - 15 ug of Arabidopsis thaliana sun1 sun2 mutant callus total protein
MW markers: PageRuler™ Plus Prestained Protein Ladder, 10 to 250 kDa (ThermoFisherScientific)15 µg/well of total protein extracted freshly from Arabidopsis thaliana callus with 50 mM Tris pH7.5, 150 mM NaCl, 1 mM EGTA, 1 mM DTT, 1% TritonX-100, 0.5% sodium deoxycholate, 0.1% SDS, 1xprotease inhibitor cocktail (ThermoFisherScientific) and denatured with Bio-Rad 1x Laemmli sample buffer (BioRad) at 95°C 5 min. Samples were separated on 12% SDS-PAGE and blotted 15V, 16h to PVDF (pore size of 45 µm), using wet transfer. Blot was blocked with 5% BSA for 1h/RT agitation. Blot was incubated in the primary antibody in 2% BSA in TBS-Tat a dilution of 1: 1 000 for 1h/RT with agitation in 2% BSA with 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 Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:25 000 in TBS-T for 1h/RT with agitation. The blot was washed as above and developed for 2 min with Agrisera ECLBright. Exposure time was 10 seconds.
Courtesy of Wei Wang, Umeå Plant Science Centre, Sweden
- Background
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Background: SUN1,2 (nuclear envelope protein) is a member of the Sad1/UNC-84 (SUN)-domain proteins.SUN domain proteins are part of the cytoskeletal-nucleoskeletal bridging complexes. These proteins are localized to the nuclear envelope and are present as homomers and heteromers in vivo. Involved in maintaining the elongated nuclear shape of epidermal cells.
- Protocols
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Agrisera Western Blot protocol and video tutorials
Preparation of cytosolic and nuclear protein fractions
1. Prepare protoplasts from 50 ml Arabidopsis thaliana cell culture according to the protocol of PEG transfection.
2. Resuspend protoplasts in 10 ml GH buffer and keep the solution on ice for 10 min.
GH buffer: 100mM glycine
0.1% Hexylene glycol
0.37M (4.7% w/v) saccharose
0.3mM Spermine
1.0mM Spermidine
pH 8.3 with Ca(OH)2
3. To release nuclei add Triton X100 to a final concentration of 0.1%. Pipetting gently up and down several
times with a plastic pipette might be necessary to lyse cells.
4. After 5min sediment nuclei by centrifugation at 1000 xg for 15 min at 4°C. Save supernatant as the
cytoplasmic fraction. Wash the pelleted nuclei two times with GHT (GH+0.1% TX100) then finally
resuspended in a suitable volume of extraction buffer + protease inhibitors.Courtesy Dr. Laszlo Bako, Umeå Plant Science Centre, Sweden
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