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RabMAb

Anti-Sumo 1 抗体 [Y299] - ChIP Grade (ab32058)

製品の概要

  • 製品名
    Anti-Sumo 1 antibody [Y299] - ChIP Grade
    Sumo 1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [Y299] to Sumo 1 - ChIP Grade
  • 特異性
    ab32058 recognises small ubiquitin-related modifier-1 (SUMO-1), also known as SMT3, Sentrin, GMP1 UBL1 and PIC1.
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IF, Flow Cyt, IP, ChIPmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues near the C-term of human SUMO-1.

  • ポジティブ・コントロール
    • WB: HeLa and NIH-3T3 cell lysates IHC: lung carcinoma ICC/IF: HeLa cells FC: HeLa cells
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

     

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

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32058 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB 1/1000 - 1/5000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).
IHC-P 1/250.
ICC/IF 1/250 - 1/500.
Flow Cyt 1/20 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/20.
ChIP Use 5 µg for 25 µg of chromatin.

ターゲット情報

  • 機能
    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.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the ubiquitin family. SUMO subfamily.
    Contains 1 ubiquitin-like domain.
  • 翻訳後修飾
    Cleavage of precursor form by SENP1 or SENP2 is necessary for function.
    Polymeric SUMO1 chains undergo polyubiquitination by RNF4.
  • 細胞内局在
    Nucleus membrane. Nucleus speckle. Cytoplasm. Recruited by BCL11A into the nuclear body.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-Sumo 1 antibody [Y299] - ChIP Grade 画像

  • 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.

  • 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.

  • 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)

  • ab32058 immunoprecipitating Sumo 1. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP secondary antibody (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

  • 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).

  • 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.

  • 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.

  • 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.

  • All lanes : Anti-Sumo 1 antibody [Y299] - ChIP Grade (ab32058) at 1/5000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 12 kDa

    Blocking and diluting buffer: 5% NFDM/TBST



  • Predicted band size : 12 kDa

    ab32058 was shown to specifically react with Sumo 1 when Sumo 1 knockout samples were used. Wild-type and Sumo 1 knockout samples were subjected to SDS-PAGE. Ab32058 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • 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.

  • All lanes : Anti-Sumo 1 antibody [Y299] - ChIP Grade (ab32058) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : NIH 3T3 cell lysate


    Predicted band size : 12 kDa
    Observed band size : 12,80 kDa (why is the actual band size different from the predicted?)
  • 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.

  • IHC of paraffin-embedded human lung carcinoma using anti-SUMO 1 (ab32058) diluted 1:250
  • Immunofluorescent staining of HeLa cells using anti-SUMO 1 (ab32058) diluted 1/250.

Anti-Sumo 1 antibody [Y299] - ChIP Grade (ab32058) 使用論文

This product has been referenced in:
  • Yang WS  et al. SUMO modification of a heterochromatin histone demethylase JMJD2A enables viral gene transactivation and viral replication. PLoS Pathog 13:e1006216 (2017). WB ; Human . Read more (PubMed: 28212444) »
  • Cloutier JM  et al. Mammalian meiotic silencing exhibits sexually dimorphic features. Chromosoma 125:215-26 (2016). Read more (PubMed: 26712235) »

See all 40 Publications for this product

Product Wall

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Spinal cord)
Permeabilization
Yes - 0.1% Triton-X
Specification
Spinal cord
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 21 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Permeabilization
Yes - 0.5%
Specification
U2OS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jul 01 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (breast cancer cell lines)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
breast cancer cell lines
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jul 01 2016

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

投稿 Aug 20 2012

I have issued a free of charge replacement for one vial of ab32058 with the order number 1145601.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of c...

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Thanks for your reply.

I am sorry for the confusion. In this case I am happy to send you 1 additional vial of ab32058 free of charge to make up for the difference in mass between the vial of ab32058 you purchased 2 years ago and the vial you ...

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Rabbit monoclonal antibodies do get titered based on our WB testing for each lot.
In certain cases, the antibody concentration is adjusted in order to be in the acceptable
range for WB dilution factor (in this case, 1:1000 - 10,000).
We ...

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Thank you for your enquiry.

The concentration of our current lot of ab32058, lot GR44178, is 0.092 mg/ml.

I hope this is helpful. Please contact us again if you have any further questions.

Thank you for contacting us.

We can provide blocking peptide for this product the price will be £113 for 100ug size. Could you let us know if you are happy purchasing this then I will inform datasheet for publishing the datasheet on webs...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (LIVER)
Specification
LIVER
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37%FORMALDEHYDE
Detection step
Semiquantitative PCR
Positive control
POL11
Negative control
IgG
Username

Mr. Natarajan Balasubramaniyan

Verified customer

投稿 May 19 2012

1-10 of 14 Abreviews or Q&A

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

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