SUMO3 | Small ubiquitin-like modifier protein 3 (peptide antibody)

Product no: AS08 349

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

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  • Product Info
  • Immunogen:

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

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 g
    Reconstitution: For reconstitution add 50 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; 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: Western blot (WB)
    Recommended dilution: 1 : 5000 for detection of recombinant SUMO3 (WB)
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • 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 chemiluminescent detection reagent, according to the manufacturer's instructions. Exposure time was 15 second.

    Courtesy of Dr.  Concepción Almoguera, CSIC, Spain
  • Additional Information
  • Additional information (application): The antibody is recognizing recombinant SUMO3 and shows no cross reactivity to SUMO1/2
  • Background
  • 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.
  • Product Citations
  • 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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

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