Overview

  • Product name

    Anti-Cyclin T1 antibody [EPR17982]
    See all Cyclin T1 primary antibodies
  • Description

    Rabbit monoclonal [EPR17982] to Cyclin T1
  • Host species

    Rabbit
  • Specificity

    Note that the antibody detects the target protein from human cell lysates but not tissue lysates.
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Cyclin T1 aa 250-550. The exact sequence is proprietary.
    Database link: O60563

  • Positive control

    • WB: K562, Jurkat, HeLa, HepG2, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and spleen lysates. IHC-P: Mouse liver and rat kidney tissues. ICC/IF: K562, NIH/3T3 and PC-12 cells. Flow Cyt: NIH/3T3 cells. IP: PC-12 whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab184703 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
WB 1/1000. Detects a band of approximately 81 kDa (predicted molecular weight: 81 kDa).
ICC/IF 1/500.
Flow Cyt 1/120.
IP 1/40.
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

The IHC application is recommended for mouse and rat only.

Target

  • Function

    Regulatory subunit of the cyclin-dependent kinase pair (CDK9/cyclin-T1) complex, also called positive transcription elongation factor B (P-TEFb), which is proposed to facilitate the transition from abortive to productive elongation by phosphorylating the CTD (carboxy-terminal domain) of the large subunit of RNA polymerase II (RNA Pol II). In case of HIV or SIV infections, binds to the transactivation domain of the viral nuclear transcriptional activator, Tat, thereby increasing Tat's affinity for the transactivating response RNA element (TAR RNA). Serves as an essential cofactor for Tat, by promoting RNA Pol II activation, allowing transcription of viral genes.
  • Tissue specificity

    Ubiquitously expressed.
  • Sequence similarities

    Belongs to the cyclin family. Cyclin C subfamily.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • CCN T1 antibody
    • CCNT antibody
    • CCNT 1 antibody
    • Ccnt1 antibody
    • CCNT1_HUMAN antibody
    • CDK9 associated C type protein antibody
    • Cyc T1 antibody
    • Cyclin C related protein antibody
    • Cyclin T antibody
    • cyclin T1 antibody
    • Cyclin T1b antibody
    • Cyclin-T antibody
    • Cyclin-T1 antibody
    • CYCT 1 antibody
    • CycT1 antibody
    • HIVE1 antibody
    • Human immunodeficiency virus 1 expression antibody
    • Human immunodeficiency virus type 1 (HIV 1) expression (elevated) 1 antibody
    • pTEFb subunit antibody
    • Subunit of positive elongation transcription factor b antibody
    see all

Images

  • All lanes : Anti-Cyclin T1 antibody [EPR17982] (ab184703) at 1/1000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 81 kDa
    Observed band size: 81 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Ab184703 staining Cyclin T1 in Jurkat (human T cell leukemia T lymphocyte). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1/100 dilution (6.4µg/ml). An Alexa Fluor® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). Ab195888, anti-alpha Tubulin [DM1A] – Microtubule Marker (Alexa Fluor® 594) was used as counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing nuclear staining in Jurkat cell line.

  • Ab184703 staining Cyclin T1 in MCF7 (human breast adenocarcinoma epithelial cell). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1:100 dilution (6.4 μg/ml). An Alexa Fluor® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). Ab195888, anti-alpha Tubulin [DM1A] – Microtubule Marker (Alexa Fluor® 594) was used as counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing nuclear staining in MCF7 cell line.

  • All lanes : Anti-Cyclin T1 antibody [EPR17982] (ab184703) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 81 kDa
    Observed band size: 81 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2 and 3: 15 seconds; Lane 4: 3 minutes.

    The antibody did not detect the target protein from human tissues (WB or IHC) but the IHC application is recommended for mouse and rat. In addition the failure of human tissue WB might result from shortage of proper human tissue.

  • All lanes : Anti-Cyclin T1 antibody [EPR17982] (ab184703) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 81 kDa
    Observed band size: 81 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2, 3 and 4: 15 seconds.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cyclin T1 with ab184703 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on hepatocytes of mouse liver is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Cyclin T1 with ab184703 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on rat kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Cyclin T1 with ab184703 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR17982] -  Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Cyclin T1 was immunoprecipitated from 1mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate with ab184703 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184703 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: PC-12 whole cell lysate 10µg (Input).
    Lane 2: ab184703 IP in PC-12 whole cell lysate.
    Lane 3: Rabbit IgG,monoclonal [EPR17982] - Isotpe Control (ab172730) instead of ab184703 in PC-12 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

References

ab184703 has not yet been referenced specifically in any publications.

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