Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

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Overview

  • Product name

    Anti-Sumo 1 antibody [Y299] - BSA and Azide free
    See all Sumo 1 primary antibodies

  • Description

    Rabbit monoclonal [Y299] to Sumo 1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, ICC/IF, WB, IHC-P, ChIPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Sumo 1 aa 50-150 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa and NIH-3T3 cell lysates IHC: lung carcinoma ICC/IF: HeLa cells FC: HeLa cells

  • General notes

    Ab219724 is the carrier-free version of ab32058. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab219724 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab219724 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
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.

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

  • Other - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Other - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)Image from Maroui M.A. et al Send to PLoS Pathog. 2016 Sep 12;12(9):e1005834. doi: 10.1371/journal.ppat.1005834. eCollection 2016 Sep.

    HSV-1 MA pattern corresponds to vDCP-NBs and contains SUMO proteins

    Immuno-DNA-FISH showing HSV-1 genomes (red), small ubiquitin modifier (SUMO) proteins (green), and cellular chromatin (DAPI, blue/grey).  SUMO-1 in (i) non-infected neurons, (ii) RC-containing neurons and (iii, iv) MA/vDCP-NBs or S/vDCP-NB-containing neurons.

    (From Figure 2C of Maroui et al)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • ChIP - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    ChIP - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    Chromatin was prepared from SK-OV-3 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32058 (blue), and 20µl of Anti rabbit IgG sepharose beads. 5μg of rabbit normal IgG was added to the beads control (yellow).  The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    ChIP was performed according to the literature (PMID: 23770046).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunoprecipitation - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunoprecipitation - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 immunoprecipitating Sumo 1. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

    Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10ug
    Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32058 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Flow Cytometry - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Flow Cytometry - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a counterstain for primary antibody ab133645 at 1/200. DAPI was used as a nuclear counterstain and PBS as a negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in rat stomach tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in human bladder carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    ab32058 staining Sumo 1 in human endometrium tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Flow Cytometry - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Flow Cytometry - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Sumo 1 with ab32058 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    IHC of paraffin-embedded human lung carcinoma using anti-SUMO 1 (ab32058) diluted 1:250

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)

    Immunofluorescent staining of HeLa cells using anti-SUMO 1 (ab32058) diluted 1/250.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

  • Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)
    Immunocytochemistry/ Immunofluorescence - Anti-Sumo 1 antibody [Y299] - BSA and Azide free (ab219724)Image from Cuchet-Louren?o D et al. PLoS Pathog. 2011 Jul;7(7):e1002123. Epub 2011 Jul 14. Fig 9.; doi:10.1371/journal.ppat.1002123; July 14 2011 PLoS Pathog 7(7): e1002123.

    Immunofluorescence analysis of ICP0-null mutant HSV-1 infected HepaRG cells, staining Sumo1 (green) with ab32058. An AlexaFluor®-conjugated goat anti-rabbit IgG was used as the seconday antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32058).

References

ab219724 has not yet been referenced specifically in any publications.

Publishing research using ab219724? Please let us know so that we can cite the reference in this datasheet.

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