SUMO3 | Small ubiquitin-like modifier protein 3

AS08 349 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

SUMO3 | Small ubiquitin-like modifier protein 3 in the group Plant/Algal Antibodies / Developmental Biology / Signal transduction at Agrisera AB (Antibodies for research) (AS08 349)


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product information
Background SUMO3 (Small ubiquitin-related modifier 3) is a small polypeptide that becomes covalently attached to various intracellular proteins leading to their post-translational modification. 

KLH-conjugated peptide derived from SUMO3 sequence of Arabidopsis thaliana, UniProt: Q9FLP5, TAIR: AT5G55170

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized in PBS pH 7.4
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl 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)
Related products

AS08 308 |anti-SUMO1 antibodies

Additional information
application information
Recommended dilution 1 : 5000 for detection of recombinant SUMO3 (WB)
Expected | apparent MW
Confirmed reactivity Arabidopsis thaliana
Predicted reactivity
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information The antibody is recognizing recombinant SUMO3 and shows no cross reactivity to SUMO1/2.
Selected references Saleh et al. (2015). Posttranslational Modifications of the Master Transcriptional Regulator NPR1 Enable Dynamic but Tight Control of Plant Immune Responses. Cell Host Microbe. 2015 Aug 12;18(2):169-82. doi: 10.1016/j.chom.2015.07.005.

application example

Western blot using anti-SUMO3 antibodies

Protein, tagged with a 6xhis-tag, was expressed from a strain of E. coli that reconstituted the sumoylation system with SUMO3 (described in Elrouby et al). Cells were harvested and lysated by sonication in buffer containing 6M urea. Proteins were used to load a Nickel column under denaturing conditions with buffers containing 6M urea. A pH gradient was applied to release the specifically bound proteins and 0.5 ml aliquots were collected. The presence of the desired protein in each aliquot was checked in SDS-PAGE by Comassie staining. 3 ul of the aliquot showing the most intense signal were separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with TBS-T (0.1%) and 5% dry-milk powder for 4 h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 overnight at 4ºC with agitation. The antibody solution was decanted and the blot was rinsed 4 times for 15 min in TBS-T (0.25%) at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602 from Agrisera ) diluted to 1:50 000 in TBS-T (0.1%) and 5% dry-milk powder for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Advance according to the manufacturer's instructions. Exposure time was 15 second.

Courtesy of Dr.  Concepción Almoguera, CSIC, Spain

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