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SUN1,2 (nuclear envelope protein) (Arabidopsis thaliana)

AS15 2856 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

SUN1,2 (nuclear envelope protein) (Arabidopsis thaliana) in the group Antibodies for Plant/Algal  / Compartment Markers / Nuclear marker at Agrisera AB (Antibodies for research) (AS15 2856)

DATA SHEET IN PDF

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How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from SUN1,2 sequences of Arabidopsis thaliana SUN1 UniProt: Q9FF75, TAIR: AT5G04990, SUN2, UniProt: Q9SG79, TAIR: AT3G10730

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
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), Immunocytochemistry (ICC)
Recommended dilution 1 : 1000 (WB), 1 : 200 (ICC)
Expected | apparent MW

51.5 kDa (SUN1); 49.9 kDa (SUN2)

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arabidopsis thaliana
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Application example

western blot using anti-SUN1,2 antibodies

Chloroplast fraction was obtained using Chloroplast Isolation Kit (CPISO-1KT, Sigma-Aldrich). 1. Supernatant proteins after DNA digestion, 2. Unquantified, 10 µl of pellet after DNA digestion. Signal of SMT1 (Sterol methyltransferase 1, marker antibody of integral ER membrane) was not detected. Nuclei fraction was isolated from 1 g fresh weight of 3-4 old day cell suspension culture of Arabidopsis thaliana Ler according to protocol published by Xu and Copeland (2012). Nuclei fraction was stored in NSB buffer (with added ROCHE and PMSF inhibitors) in 80 °C. Residue of nuclei was centrifuged and pellet was re-suspended in 100 µl of modified RIPA buffer with added protease inhibitors and incubated 1h on ice. Then 20 µl of DNase I was added into each sample and all samples were incubated on ice for 45 min. After that 45 min another 20 µl of DNase I was added and incubated under the same conditions. Samples were twice centrifuged 1) 15 000 × g for 15 min and 2) 30 000 × g for 30 min. Pellets from the first step of centrifugation was stored. Proteins in second supernatant were precipitated with ice-cold acetone over night and twice washed. Protein pellet was dissolved in buffer contained 6 M Urea, 2 M Thiourea and 25 mM Tris, pH = 7,5. Proteins were quantified by using Bradford method. Totally 3 µg of protein was loaded on gel. Pellet after DNase treatment was dissolved in 20 µl of LSB buffer. Samples were boiled on 95°C for 10 min before loading on 4 / 12% polyacrylamide gel. Electrophoretic conditions: 90 V during whole separation (2 h); Transfer conditions: 90 mA, 30 V over night, nitrocellulose membrane; Blocking: 1 h, 5% low fat milk; Primary antibody incubation: 1 h at a dilution of 1: 1000; Washes: 3 × 5 min.; Secondary antibody: 1 h at a dilution of 1: 5000; washes: 3 × 10 min. ELC substrate was loaded on membrane and signals were detected with ChemiDoc system (Bio-Rad).

Courtesy of Dr. Martin Kubeš, Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Chemical Biology and Genetics, Olomouc, Czech Republic and Vladimír Skalický, Laboratory of Growth Regulators, Faculty of Science of Palacký University and Institute of Experimental Botany ASCR, Olomouc, Czech republic

Additional information

For other species use product AS15 2857
This antibody is not suitable for immunolocalization.

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collection of antibodies to DNA/RNA/cell cycle

Plant and algal protein extraction buffer

Background

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.

Product citations

Selected references Wang et al. (2019) OPENER Is a Nuclear Envelope and Mitochondria Localized Protein Required for Cell Cycle Progression in Arabidopsis. Plant Cell. 2019 Jul;31(7):1446-1465. doi: 10.1105/tpc.19.00033

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