Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-HIP2/LIG 抗体 [EP1145Y] (ab52930)

Reviews (1)Q&A (1)References (3)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
  • Anti-HIP2/LIG antibody [EP1145Y] (ab52930)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1145Y] to HIP2/LIG
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-HIP2/LIG antibody [EP1145Y]
    HIP2/LIG 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EP1145Y] to HIP2/LIG

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-Pmore details
    適用なし: Flow Cyt or ICC/IF

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Rat

  • 免疫原

    Synthetic peptide within Human HIP2/LIG aa 150-250 (internal sequence). The exact sequence is proprietary.

  • ポジティブ･コントロール

    • WB: HCT116, HeLa, Jurkat and Daudi cell lysates. IHC-P: Human liver tissue.

  • 特記事項

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab52930の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/100000. Detects a band of approximately 24 kDa (predicted molecular weight: 22 kDa).
IHC-P (1)
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
特記事項
WB
1/100000. Detects a band of approximately 24 kDa (predicted molecular weight: 22 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
追加情報
Is unsuitable for Flow Cyt or ICC/IF.

ターゲット情報

  • 機能

    Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro, in the presence or in the absence of BRCA1-BARD1 E3 ubiquitin-protein ligase complex, catalyzes the synthesis of 'Lys-48'-linked polyubiquitin chains. Does not transfer ubiquitin directly to but elongates monoubiquitinated substrate protein. Mediates the selective degradation of short-lived and abnormal proteins, such as the endoplasmic reticulum-associated degradation (ERAD) of misfolded lumenal proteins. Ubiquitinates huntingtin. May mediate foam cell formation by the suppression of apoptosis of lipid-bearing macrophages through ubiquitination and subsequence degradation of p53/TP53. Proposed to be involved in ubiquitination and proteolytic processing of NF-kappa-B; in vitro supports ubiquitination of NFKB1. In case of infection by cytomegaloviruses may be involved in the US11-dependent degradation of MHC class I heavy chains following their export from the ER to the cytosol. In case of viral infections may be involved in the HPV E7 protein-dependent degradation of RB1.

  • 組織特異性

    Expressed in all tissues tested, including spleen, thymus, prostate, testis, ovary, small intestine, colon, peripheral blood leukocytes, T-lymphocytes, monocytes, granulocytes and bone marrow mononuclear cells. Highly expressed in brain, with highest levels found in cortex and striatum and at lower levels in cerebellum and brainstem.

  • パスウェイ

    Protein modification; protein ubiquitination.

  • 配列類似性

    Belongs to the ubiquitin-conjugating enzyme family.
    Contains 1 UBA domain.

  • 翻訳後修飾

    Sumoylation at Lys-14 impairs catalytic activity.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • E2 25K antibody
    • E2(25K) antibody
    • HIP-2 antibody
    • Huntingtin interacting protein 2 antibody
    • Huntingtin-interacting protein 2 antibody
    • HYPG antibody
    • LIG antibody
    • UBC1 antibody
    • UBE2K antibody
    • UBE2K_HUMAN antibody
    • Ubiquitin carrier protein antibody
    • Ubiquitin conjugating enzyme E2 25 kDa antibody
    • ubiquitin conjugating enzyme E2K antibody
    • ubiquitin conjugating enzyme E2K (UBC1 homolog, yeast) antibody
    • Ubiquitin protein ligase antibody
    • Ubiquitin-conjugating enzyme E2 K antibody
    • Ubiquitin-conjugating enzyme E2(25K) antibody
    • Ubiquitin-conjugating enzyme E2-25 kDa antibody
    • Ubiquitin-conjugating enzyme E2-25K antibody
    • Ubiquitin-protein ligase antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)

    Ab52930 (1:250) staining human HIP2/LIG in human liver tissue by immunohistochemistry using paraffin embedded tissue.

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

  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    All lanes : Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : UBE2K knockout HCT116 cell lysate
    Lane 3 : Daudi cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 22 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab52930 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab52930 Anti-HIP2/LIG antibody [EP1145Y] was shown to specifically react with HIP2/LIG in wild-type HCT116 cells. Loss of signal was observed when knockout cell line ab266899 (knockout cell lysate ab257779) was used. Wild-type and HIP2/LIG knockout samples were subjected to SDS-PAGE. ab52930 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    All lanes : Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : UBE2K knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 22 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab52930 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab52930 Anti-HIP2/LIG antibody [EP1145Y] was shown to specifically react with HIP2/LIG in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266031 (knockout cell lysate ab257778) was used.  Wild-type and HIP2/LIG knockout samples were subjected to SDS-PAGE.  ab52930 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.  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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Western blot - Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Anti-HIP2/LIG antibody [EP1145Y] (ab52930) at 1/100000 dilution + Daudi cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled at 1/2000 dilution

    Predicted band size: 22 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?

  • Anti-HIP2/LIG antibody [EP1145Y] (ab52930)
    Anti-HIP2/LIG antibody [EP1145Y] (ab52930)

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (3)

ab52930 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab52930 は 3 報の論文で使用されています。

  • McManus FP  et al. Quantitative SUMO proteomics reveals the modulation of several PML nuclear body associated proteins and an anti-senescence function of UBC9. Sci Rep 8:7754 (2018). PubMed: 29773808
  • Su J  et al. Reduction of HIP2 expression causes motor function impairment and increased vulnerability to dopaminergic degeneration in Parkinson's disease models. Cell Death Dis 9:1020 (2018). PubMed: 30282965
  • Benedikt A  et al. The leukemogenic AF4-MLL fusion protein causes P-TEFb kinase activation and altered epigenetic signatures. Leukemia 25:135-44 (2011). WB ; Human . PubMed: 21030982

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HIP2 antibody [EP1145Y]

 Good
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 10% Tween-20
Specification
Brain
Blocking step
Heat-Inactivated Normal Donkey Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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.

Abcam user community

Verified customer

投稿 Jan 22 2016

Question

Are there any known issues with WB analysis using the Lys 14 -SUMOylated E2-25K? We are conducting WB assays to determine the level of SUMOylation of this substrate and the SUMOylated E2-25K appears much fainter than the unmodified E2-25K. Is the epitope located around Lys14 of E2-25K?

Read More
Answer

The antibody ab52930 is raised against a peptide located within the range of amino acids 140 - 170 of human HIP2/E2-25K, http://www.uniprot.org/uniprot/P61086. There are no known issues of detection of SUMOylated samples. I am not sure what is happening that would cause the signal to be fainter, unless there is some modification or degradation at or near the epitope that the antibody recognizes that only exists in the samples that are SUMOylated at Lys 14.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com