Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Sumo 1 antibody [EP298] (ab133352)

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Overview

  • Product name

    Anti-Sumo 1 antibody [EP298]
    See all Sumo 1 primary antibodies

  • Description

    Rabbit monoclonal [EP298] to Sumo 1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Sumo 1 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB- HeLa whole cell lysate (ab150035), MCF-7 whole cell lysate, A549 whole cell lysate, NIH/3T3 whole cell lysate, C6 whole cell lysate, HL-60 cell lysate and A431 cell lysate. IHC- Human papillary cell thyroid gland carcinoma tissue, Human Breast carcinoma tissue, Human Glioma tissue, Human Urinary bladder transitional carcinoma tissue and Human tonsil tissue.

  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133352 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
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/10000. Predicted molecular weight: 11.5 kDa.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Ubiquitin-like protein that can be covalently attached to proteins as a monomer or a lysine-linked polymer. Covalent attachment via an isopeptide bond to its substrates requires prior activation by the E1 complex SAE1-SAE2 and linkage to the E2 enzyme UBE2I, and can be promoted by E3 ligases such as PIAS1-4, RANBP2 or CBX4. This post-translational modification on lysine residues of proteins plays a crucial role in a number of cellular processes such as nuclear transport, DNA replication and repair, mitosis and signal transduction. Involved for instance in targeting RANGAP1 to the nuclear pore complex protein RANBP2. Polymeric SUMO1 chains are also susceptible to polyubiquitination which functions as a signal for proteasomal degradation of modified proteins. May also regulate a network of genes involved in palate development.

    • Involvement in disease

      Defects in SUMO1 are the cause of non-syndromic orofacial cleft type 10 (OFC10) [MIM:613705]; also called non-syndromic cleft lip with or without cleft palate 10. OFC10 is a birth defect consisting of cleft lips with or without cleft palate. Cleft lips are associated with cleft palate in two-third of cases. A cleft lip can occur on one or both sides and range in severity from a simple notch in the upper lip to a complete opening in the lip extending into the floor of the nostril and involving the upper gum. Note=A chromosomal aberation involving SUMO1 is the cause of OFC10. Translocation t(2;8)(q33.1;q24.3). The breakpoint occurred in the SUMO1 gene and resulted in haploinsufficiency confirmed by protein assays.

    • Sequence similarities

      Belongs to the ubiquitin family. SUMO subfamily.
      Contains 1 ubiquitin-like domain.

    • Post-translational
      modifications

      Cleavage of precursor form by SENP1 or SENP2 is necessary for function.
      Polymeric SUMO1 chains undergo polyubiquitination by RNF4.

    • Cellular localization

      Nucleus membrane. Nucleus speckle. Cytoplasm. Recruited by BCL11A into the nuclear body.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • DAP1 antibody
      • GAP modifying protein 1 antibody
      • GAP-modifying protein 1 antibody
      • GMP 1 antibody
      • GMP1 antibody
      • OFC10 antibody
      • PIC 1 antibody
      • PIC1 antibody
      • SENP2 antibody
      • Sentrin 1 antibody
      • Sentrin antibody
      • Small ubiquitin related modifier 1 antibody
      • Small ubiquitin-like modifier 1 antibody
      • Small ubiquitin-related modifier 1 antibody
      • SMT3 antibody
      • SMT3 homolog 3 antibody
      • SMT3 suppressor of mif two 3 homolog 1 antibody
      • SMT3, yeast, homolog 3 antibody
      • Smt3C antibody
      • SMT3H3 antibody
      • SUMO-1 antibody
      • SUMO1 antibody
      • SUMO1_HUMAN antibody
      • Ubiquitin homology domain protein PIC1 antibody
      • Ubiquitin Like 1 antibody
      • Ubiquitin like protein SMT3C antibody
      • Ubiquitin like protein UBL1 antibody
      • Ubiquitin-homology domain protein PIC1 antibody
      • Ubiquitin-like protein SMT3C antibody
      • Ubiquitin-like protein UBL1 antibody
      • UBL 1 antibody
      • UBL1 antibody
      see all

    Images

    • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
      Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Sumo 1 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: MCF-7 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab133352 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab133352 was shown to recognize Sumo 1 when Sumo 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Sumo 1 knockout samples were subjected to SDS-PAGE. ab133352 and ab8245 (loading control to GAPDH) were diluted 1/000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical staining of paraffin-embedded human thyroid carcinoma sections labelling Sumo 1 with purified ab133352 at dilution of 1:100. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

    • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
      Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
      All lanes : Anti-Sumo 1 antibody [EP298] (ab133352) at 1/10000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
      Lane 2 : MCF-7 (human breast carcinoma) whole cell lysate
      Lane 3 : A549 (human lung carcinoma) whole cell lysate
      Lane 4 : NIH/3T3 (mouse embryo) whole cell lysate
      Lane 5 : C6 (rat glioma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size: 11.5 kDa
      Observed band size: 18-90 kDa
      why is the actual band size different from the predicted?



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
      Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
      All lanes : Anti-Sumo 1 antibody [EP298] (ab133352) at 1/1000 dilution (un-purified)

      Lane 1 : HeLa cell lysate
      Lane 2 : HL60 cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : NIH3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 11.5 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical analysis of paraffin-embedded Human papillary cell thyroid gland carcinoma tissue labelling Sumo 1 with un-purified ab133352 at 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical analysis of paraffin embedded Human Breast carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical analysis of paraffin embedded Human Glioma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical analysis of paraffin embedded Human Urinary bladder transitional carcinoma tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)

      Immunohistochemical analysis of paraffin embedded normal Human tonsil tissue labelling Sumo 1 with un-purified ab133352 at dilution of 1/100-1/250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Carmichael RE  et al. Insulin-dependent GLUT4 trafficking is not regulated by protein SUMOylation in L6 myocytes. Sci Rep 9:6477 (2019). Read more (PubMed: 31019221) »
    • Tan F  et al. Attenuated SUMOylation of sirtuin 1 in premature neonates with bronchopulmonary dysplasia. Mol Med Rep 17:1283-1288 (2018). Read more (PubMed: 29115559) »
    See all 6 Publications for this product

    Publishing research using ab133352? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Sumo 1 antibody [EP298]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HAP1)
    Permeabilization
    Yes - 0.5% Triton 5 minutes
    Specification
    HAP1
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 15 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Sumo 1 antibody [EP298]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer ph6 20 min at 97ºC IN A PT LINK
    Permeabilization
    No
    Specification
    skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 22 2015

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