Overview

  • Product name

    Anti-UBE2I / UBC9 antibody [EP2938Y]
    See all UBE2I / UBC9 primary antibodies
  • Description

    Rabbit monoclonal [EP2938Y] to UBE2I / UBC9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, ChIP, WB, IP, IHC-P, ICC, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human UBE2I/ UBC9 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • U937, HeLa, Jurkat or HUVEC cell lysates. Human brain 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 ab75854 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-Fr Use a concentration of 1 µg/ml. PubMed: 22016779
ChIP Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
IP Use at an assay dependent concentration.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC 1/100 - 1/250.
Flow Cyt 1/50.

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

Target

  • Function

    Accepts the ubiquitin-like proteins SUMO1, SUMO2, SUMO3 and SUMO4 from the UBLE1A-UBLE1B E1 complex and catalyzes their covalent attachment to other proteins with the help of an E3 ligase such as RANBP2 or CBX4. Necessary for sumoylation of FOXL2 and KAT5. Essential for nuclear architecture and chromosome segregation.
  • Tissue specificity

    Expressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus.
  • Pathway

    Protein modification; protein sumoylation.
  • Sequence similarities

    Belongs to the ubiquitin-conjugating enzyme family.
  • Cellular localization

    Nucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body.
  • Information by UniProt
  • Database links

  • Alternative names

    • C358B7.1 antibody
    • p18 antibody
    • SUMO 1 protein ligase antibody
    • SUMO conjugating enzyme UBC9 antibody
    • SUMO-conjugating enzyme UBC9 antibody
    • SUMO-protein ligase antibody
    • SUMO1 protein ligase antibody
    • UBC9 antibody
    • UBC9_HUMAN antibody
    • UBCE9 antibody
    • Ube2i antibody
    • Ubiquitin carrier protein 9 antibody
    • Ubiquitin carrier protein antibody
    • Ubiquitin carrier protein I antibody
    • Ubiquitin conjugating enzyme 9 antibody
    • Ubiquitin conjugating enzyme E2I (homologous to yeast UBC9) antibody
    • Ubiquitin conjugating enzyme E2I (UBC9 homolog, yeast) antibody
    • Ubiquitin conjugating enzyme UbcE2A antibody
    • Ubiquitin like protein SUMO 1 conjugating enzyme antibody
    • Ubiquitin protein ligase E2I antibody
    • Ubiquitin-conjugating enzyme E2 I antibody
    • Ubiquitin-protein ligase I antibody
    see all

Images

  • All lanes : Anti-UBE2I / UBC9 antibody [EP2938Y] (ab75854) at 1/10000 dilution

    Lane 1 : U937 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HUVEC cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/1000 dilution

    Predicted band size: 18 kDa
    Observed band size: 18 kDa

  • ab75854, at 1/100 dilution, staining UBE2I / UBC9 in human brain, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • UBE2I / UBC9 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to UBE2I / UBC9 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75854.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 18kDa: UBE2I / UBC9; non specific - 60kDa: We are unsure as to the identity of this extra band.
  • ab75854 staining UBE2I / UBC9 (red) in cerebral cortex of Ubc9 transgenic mice, by Immunohistochemistry (Frozen sections). Left panel shows all layers of cerebral cortex, and right panel shows an enlarged region of the Layer III-External pyramidal area.

    Samples were incubated with primary antibody at 1 µg/ml and an AlexaFluor®700-conjugated goat anti-rabbit IgG (1 µg/ml) was used as the secondary antibody.
  • Overlay histogram showing HepG2 cells stained with ab75854 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75854, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:

  • Gao K  et al. SUMO peptidase ULP-4 regulates mitochondrial UPR-mediated innate immunity and lifespan extension. Elife 8:N/A (2019). Read more (PubMed: 30642431) »
  • 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) »
See all 18 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Jun 14 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
20 µg
Specification
MEF
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 03 2019

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Spinal cord 16 micron cryosection)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
Permeabilization
Yes - 0.5% Triton X-100
Specification
Spinal cord 16 micron cryosection
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 08 2014

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Hela)
Total protein in input
100 µg
Immuno-precipitation step
Protein A/G
Specification
Hela

Abcam user community

Verified customer

Submitted Aug 27 2012

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

Mr. Natarajan Balasubramaniyan

Verified customer

Submitted May 19 2012

Question

Dear Dr Keith Yes,I want use in the beginning ab32219 and ab32058 for the ChiP assay in human liver cell line.Based on the results I want use other antibody as well.I am going to order all the antibody now.Thanks. With regards Bala ________________________________________ From: technical@abcam.com [technical@abcam.com] Sent: Thursday, January 12, 2012 3:27 PM To: Balasubramaniyan, Natarajavelalar Subject: Reply from Abcam to your enquiry regarding ab32058, ab32219, ab75854, ab81371 [CCE3421816] [https://www.abcam.com/ps/cms/emails/CRM-Header.jpg] Dear Sir/Madam We have an answer to your inquiry: Thank you very much for your interest in ab32219, ab32058, ab81371, ab75854. To our knowledge, these products has not been tested in ChIP on human. Therefore, I can offer a discount off a future purchase if you buy now, test in ChIP on human and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test any or each of these products in ChIP on human. I will then send a discount code. This code must be issued before purchasing so please wait for my reply before ordering. 2. Purchase the antibody or antibodies either by phone, fax, or online (www.abcam.com). 3. Test it in ChIP. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for ChIP, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount. Help us improve our service. Rate your experience with us today. Your original inquiry to Abcam: Will these work on ChIP as well? Best regards, Keith Keith Beadle Scientific Support Specialist Abcam Inc. www.abcam.com Abcam Customer Services and Scientific Support Team www.abcam.com/technical [CCE3421816] Discover more at abcam.com

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Answer

Ab32058 DISCOUNT CODE: Expiration date: Ab32058 DISCOUNT CODE: Expiration date: Ab81371 DISCOUNT CODE: Expiration date: 14 May 2012 Ab75854 DISCOUNT CODE: Expiration date: I am very pleased to hear you would like to accept our offer and testthese productsinChIP. This code will give you 1 free PRIMARY ANTIBODYper codebefore the expiration date. To redeem this offer, please submit an Abreview foreach product in ChIPand include the associatedcode in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you very much for your interest in ab32219, ab32058, ab81371, ab75854. To our knowledge, these products has not been tested in ChIP on human. Therefore, I can offer a discount off a future purchase if you buy now, test inChIP on human and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and testany or each of these productsin ChIP on human. I will then send a discount code. This code must be issued before purchasing so please wait for my reply before ordering. 2. Purchasethe antibody or antibodieseither by phone, fax, or online (www.abcam.com). 3. Test it in ChIP. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifthe antibodyturns out to be unsuitable for ChIP, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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