Overview

  • Product nameAnti-Sumo 2+3 antibody
    See all Sumo 2+3 primary antibodies
  • Description
    Rabbit polyclonal to Sumo 2+3
  • SpecificityRecognises 2 bands representing Sumo 2 and Sumo 3 at 15 and 18kDa in Hela Nuclear extract by Western blotting.
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human, Xenopus laevis, Chinese Hamster
    Predicted to work with: Rat, Cow, Pig, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Sumo 2+3.

    (Peptide available as ab13760.)

  • Positive control
    • WB: HeLa Nuclear cell lysate. IHC-P: Human colon adenocarcinoma tissue.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3742 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 15, 18 kDa (predicted molecular weight: 11.6 , 10.8 kDa).
IHC-P 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

Anti-Sumo 2+3 antibody images

  • Left-hand images show uninfected cells and the co-localization of SUMO-2/3 with PML (red) in control (upper rows of each block of 4 images) and PML depleted (low rows of each block of 4 images) HepaRG cells. Right-hand images show typical examples of recruitment of the indicated proteins to sites associated with HSV-1 genomes (ICP4; red) in cells at the edges of ICP0 null mutant (ΔICP0) plaques in control and PML depleted HepaRG cells. Scale bars indicate 5 µm.

    Cells on glass coverslips were fixed with 1.5% formaldehyde in PBS containing 2% sucrose then treated with 0.5% Nonidet P40 substitute in PBS/10% sucrose. SUMO-2/3 was detected with ab3742. An Alexa-conjugated anti-rabbit IgG was used as the secondary antibody.


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11.6 , 10.8 kDa


    Exposure time : 1 minute

    Lanes 1 & 3: Sumo 2+3 antibody (ab3742) at 1/500 dilution
    Lanes 2 & 4: Sumo 2+3 antibody (ab3742) at 1/1000 dilution
    Lanes 1-4: HeLa nuclear extract at 20 ug

    Lane 1: as above
    Lane 2: as above
    Lane 3: Sumo 2+3 peptide (ab13760) at 1 ug/ml
    Lane 4: Sumo 2+3 peptide (ab13760) at 1 ug/ml


    Secondary
    Goat polyclonal to Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution developed using the ECL technique

    Performed under reducing conditions.
     
    Exposure time: 1 minute

    Predicted band sizes : 11.6 & 10.

  • IHC image of Sumo 2+3 staining in human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3742, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Image courtesy of Human Protein Atlas

    ab3742 staining Sumo 2 + 3 in Human skin. The  paraffin embedded human skin tissue was incubated with ab3742 (1/800 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab3742 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • Immunofluorescent imaging of human cells (U2OS) with ab3742 confirms the specificity of this antibody.  Antibody signal is localised exclusively to the nucleus, with diffuse background staining of the nucleoplasm.  Intense foci of staining are also evident, corresponding to SUMO-2/3 accumulation in nuclear subdomains such as the PML body.  This image is in exact agreement with several published reports (see for example Saitoh H et al.). 

    IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees. Nuclei are stained with Hoechst stain.

  • ab3742 staining Sumo 2+3 in Xenopus laevis oocyte cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with NP-40 0.1% in PBS and blocked with 3% BSA for 60 minutes at 23°C. Samples were incubated with primary antibody (1/250 in PBS + 3% BSA) for 1 hour at 23°C. An undiluted Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

References for Anti-Sumo 2+3 antibody (ab3742)

This product has been referenced in:

See all 28 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Lung)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer PH 6 in a PT link 20 minutes at 96C
Permeabilization No
Specification Lung
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative Formaldehyde
Username

Miss. Virginia Gutiérrez

Verified customer

Submitted Oct 10 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEK 293T)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 50 µg
Specification HEK 293T
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Sep 11 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (12.5% SDS-PAGE gel)
Sample Human Cell lysate - whole cell (HeLa cells)
Specification HeLa cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 07 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - whole cell (Xenopus egg extract)
Loading amount 40 µg
Specification Xenopus egg extract
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Dr. Andrew Lane

Verified customer

Submitted Apr 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (Nuclei generated in oocyte extract)
Specification Nuclei generated in oocyte extract
Fixative Formaldehyde
Permeabilization Yes - PBS-0.1% NP40
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Username

Dr. Andrew Lane

Verified customer

Submitted Mar 26 2013

Thank you for contacting us.
Both ab3742 and ab81371 have immunogen with 100% homology with the mouse sequence.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Human fibroblast)
Loading amount 20 µg
Specification Human fibroblast
Gel Running Conditions Reduced Denaturing (4-12% bis-tris nupage)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 22 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (di-SUMO recombinant protein in cleavage assay)
Loading amount 0.11 µg
Specification di-SUMO recombinant protein in cleavage assay
Treatment 50nM of specific protease or 5ug HEK293T cell lysate as a positive control (0.5-15 h)
Gel Running Conditions Reduced Denaturing (4-13% Bis-Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 28 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (liver)
Loading amount 30 µg
Specification liver
Gel Running Conditions Reduced Denaturing (4-20% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 13 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (P388D1 cells (macrophages))
Loading amount 10 µg
Specification P388D1 cells (macrophages)
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Sep 23 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"