製品の概要

  • 製品名Anti-U2AF65 antibody
    U2AF65 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to U2AF65
  • アプリケーション適用あり: IHC-P, ICC/IF, IP, WBmore details
  • 種交差性
    交差種: Mouse, Human, Zebrafish
    交差が予測される動物種: Cow, Xenopus laevis
  • 免疫原

    Synthetic peptide derived from within residues 200 - 300 of Human U2AF65.

    (Peptide available as ab37529.)

  • ポジティブ・コントロール
    • This antibody gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK 293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line) MCF-7 (Human breast adenocarcinoma cell line) SHSY-5Y (Human neuroblastoma cell line) U2OS (Human osteosarcoma cell line)

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファーPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab37530 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
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration. PubMed: 21725067
WB 1/250. Detects a band of approximately 65 kDa (predicted molecular weight: 53 kDa).

ターゲット情報

  • 機能Necessary for the splicing of pre-mRNA. Induces cardiac troponin-T (TNNT2) pre-mRNA exon inclusion in muscle. Regulates the TNNT2 exon 5 inclusion through competition with MBNL1. Binds preferentially to a single-stranded structure within the polypyrimidine tract of TNNT2 intron 4 during spliceosome assembly. Required for the export of mRNA out of the nucleus, even if the mRNA is encoded by an intron-less gene. Represses the splicing of MAPT/Tau exon 10.
  • 配列類似性Belongs to the splicing factor SR family.
    Contains 3 RRM (RNA recognition motif) domains.
  • 翻訳後修飾Lysyl-hydroxylation at Lys-15 and Lys-276 affects the mRNA splicing activity of the protein, leading to regulate some, but not all, alternative splicing events.
  • 細胞内局在Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • hU2AF(65) antibody
    • hU2AF65 antibody
    • Splicing factor U2AF 65 kDa subunit antibody
    • U2 (RNU2) small nuclear RNA auxiliary factor 2 antibody
    • U2 auxiliary factor 65 kDa subunit antibody
    • U2 small nuclear ribonucleoprotein auxiliary factor (65kD) antibody
    • U2 small nuclear RNA auxiliary factor 2 antibody
    • U2 snRNP auxiliary factor large subunit antibody
    • U2af2 antibody
    • U2AF2_HUMAN antibody
    • U2AF65 antibody
    see all

Anti-U2AF65 antibody 画像

  • All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 whole cell lysate (ab7902)
    Lane 5 : HepG2 whole cell lysate (ab7900)
    Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 53 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)

    Although the predicted band size is 53kDa based on Swiss-prot data, a band of 65kDa has been previously observed.  J Biol Chem. 2004 Nov 26;279(48):49773-9 (PMID: 15377657)

  • ICC/IF image of ab37530 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab37530, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence.  5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • IHC image of ab37530 staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37530, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 10 µg
    Lane 4 : Zebrafish liver homogenate at 10 µg
    Lane 5 : Zebrafish skeletal muscle homogenate at 10 µg
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 53 kDa
    Observed band size : 53 kDa


    Exposure time : 5 minutes
  • U2AF65 was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to U2AF65 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, SHSY5Y 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 ab37530.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 65kDa: U2AF65.

Anti-U2AF65 antibody (ab37530) 使用論文

This product has been referenced in:
  • Kim JY  et al. Specificity in circadian clock feedback from targeted reconstitution of the NuRD corepressor. Mol Cell 56:738-48 (2014). Read more (PubMed: 25453762) »
  • Gu B  et al. CTD serine-2 plays a critical role in splicing and termination factor recruitment to RNA polymerase II in vivo. Nucleic Acids Res 41:1591-603 (2013). WB ; Human . Read more (PubMed: 23275552) »

See all 6 Publications for this product

Product Wall

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 gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jan 09 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Chinese Hamster Cell (CHO cells)
Specification CHO cells
Fixative Formaldehyde
Permeabilization Yes - ETOH
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
Username

Mrs. Diana Vargas

Verified customer

投稿 Feb 10 2011

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