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SGS3 | Protein suppressor of gene silencing 3

AS15 3099  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana

SGS3 | Protein suppressor of gene silencing 3 in the group Antibodies for Plant/Algal  / DNA/RNA/Cell Cycle / Transcription regulation at Agrisera AB (Antibodies for research) (AS15 3099)

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Product Information

Immunogen
KLH-conjugated peptide derived from Arabidopsis thaliana SGS3 sequence, Uniprot: Q9LDX1, TAIR: AT5G23570
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)
Recommended dilution 1: 1000 (WB)
Expected | apparent MW 72 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arabidopsis lyrata, Brassica napus, Brassica oleracea, Brassica rapa, Camelina sativa, Raphanus sativus
Species of your interest not listed? Contact us
Not reactive in

Zea mays

Application examples

Application examples

 Application example

Western blot using anti-SGS3 antibody

250 µg of grounded tissue from 2 weeks old Arabidopsis thaliana flowers from wt Col-0 and sgs3 mutant was incubated with 1 mL lysis buffer (50 mM HCl pH=7.5, 150 mM NaCl, 10% glycerol, 5mM MgCl2, 0,1% Nonidet P40, Roche protease inhibitors, 1 mM TCEP) and denatured with Laemmli buffer (2% SDS, 10% glycerol, 0.125 M Tris-HCl pH=6.8, 1% DTT) at 85°C for 7 min. The samples were separated on 12% SDS-PAGE and blotted for 60 min to nitrocelulose membrane using wet transfer. Blots were blocked with 5% milk powder  in PBST for 180 min in a cold room with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for O/N in a cold room with agitation. The antibody solution was decanted and the blot was rinsed briefly once, then washed 5 times in TBST/PBST buffer 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 for 60 min at RT with agitation. The blot was washed as above and developed for 1 min with (ECL DIY mix (100 mM Tris pH8.8, 2 mM 4IBPA (SIGMA), 1.25 mM Lumminol (SIGMA)) using X-ray film. Exposure time was 60 seconds.

Courtesy of Maria Louisa Vigh, Department of Computational and RNA Biology, University of Copenhagen, Denmark

 

Additional information

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Background

Background

Protein supressor of gene silencing 3 (SGS3) is a protein which is necessary for post-transcriptional gene silencing and virus resistance.

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