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Validated using a knockout cell lineRecombinantRabMAb

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

Reviews (2) Submit a question References (10)
Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
  • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
  • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
  • 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)
  • Western blot - Anti-Sumo 1 antibody [EP298] (ab133352)
  • 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [EP298] (ab133352)
  • Anti-Sumo 1 antibody [EP298] (ab133352)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP298] to Sumo 1
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

Applications

The Abpromise guarantee

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)
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.
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 (unpurified)

    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.

  • 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

    Lane 1 : Recombinant Human Sumo 1 protein (ab140417) at 0.01 µg
    Lane 2 : Recombinant Human Sumo 2 protein (ab140420) at 0.01 µg
    Lane 3 : Recombinant Human Sumo 3 protein (ab140414) at 0.01 µg
    Lane 4 : Recombinant Human Sumo 4 protein (ab157025) at 0.1 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 11.5 kDa
    Observed band size: 16 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking buffer: 5% NFDM/TBST

  • 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.

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

References (10)

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

ab133352 has been referenced in 10 publications.

  • Xia L  et al. SUMOylation disassembles the tetrameric pyruvate kinase M2 to block myeloid differentiation of leukemia cells. Cell Death Dis 12:101 (2021). PubMed: 33473116
  • Lin Y  et al. Hypoxia activates SUMO-1-HIF-1a signaling pathway to upregulate pro-inflammatory cytokines and permeability in human tonsil epithelial cells. Life Sci 276:119432 (2021). PubMed: 33794253
  • Clemens Böckelmann L  et al. Concomitant constitutive LNK and NFE2 mutation with loss of sumoylation in a case of hereditary thrombocythemia. Haematologica 106:1158-1162 (2021). PubMed: 32554556
  • Liu Y  et al. Expression of SUMO associated proteins in the mouse endometrium is regulated by ovarian hormones throughout the estrous cycle. Exp Ther Med 19:1855-1863 (2020). PubMed: 32104241
  • Carmichael RE  et al. Insulin-dependent GLUT4 trafficking is not regulated by protein SUMOylation in L6 myocytes. Sci Rep 9:6477 (2019). PubMed: 31019221
  • Tan F  et al. Attenuated SUMOylation of sirtuin 1 in premature neonates with bronchopulmonary dysplasia. Mol Med Rep 17:1283-1288 (2018). PubMed: 29115559
  • Liu F  et al. Overexpression of SENP1 reduces the stemness capacity of osteosarcoma stem cells and increases their sensitivity to HSVtk/GCV. Int J Oncol 53:2010-2020 (2018). PubMed: 30226577
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
  • Liu X  et al. Hypothermia inhibits the proliferation of bone marrow-derived mesenchymal stem cells and increases tolerance to hypoxia by enhancing SUMOylation. Int J Mol Med 40:1631-1638 (2017). PubMed: 29039464
  • Jester JV  et al. PPAR? Regulates Mouse Meibocyte Differentiation and Lipid Synthesis. Ocul Surf 14:484-494 (2016). PubMed: 27531629

Customer reviews and Q&As

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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
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Sep 22 2015

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