Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CBP80 antibody (ab42389)

Key features and details

  • Rabbit polyclonal to CBP80
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-CBP80 antibody
    See all CBP80 primary antibodies

  • Description

    Rabbit polyclonal to CBP80

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Xenopus laevis, Cynomolgus monkey

  • Immunogen

    Synthetic peptide corresponding to Human CBP80 aa 150-250 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab42388)

  • Positive control

    • ab42389 gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) HepG2 (Human hepatocellular liver carcinoma cell line) A431 (Human epithelial carcinoma cell line) ab42389 gave a positive result in the following nuclear lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) HepG2 (Human hepatocellular liver carcinoma cell line)

Properties

Applications

Our Abpromise guarantee covers the use of ab42389 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 Use a concentration of 1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 92 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IP Use a concentration of 5 µg/ml.

Target

  • Function

    Component of the cap-binding complex (CBC), which binds co-transcriptionally to the 5' cap of pre-mRNAs and is involved in various processes such as pre-mRNA splicing, translation regulation, nonsense-mediated mRNA decay, RNA-mediated gene silencing (RNAi) by microRNAs (miRNAs) and mRNA export. The CBC complex is involved in mRNA export from the nucleus via its interaction with THOC4/ALY, leading to the recruitment of the mRNA export machinery to the 5' end of mRNA and to mRNA export in a 5' to 3' direction through the nuclear pore. The CBC complex is also involved in mediating U snRNA and intronless mRNAs export from the nucleus. The CBC complex is essential for a pioneer round of mRNA translation, before steady state translation when the CBC complex is replaced by cytoplasmic cap-binding protein eIF4E. The pioneer round of mRNA translation mediated by the CBC complex plays a central role in nonsense-mediated mRNA decay (NMD), NMD only taking place in mRNAs bound to the CBC complex, but not on eIF4E-bound mRNAs. The CBC complex enhances NMD in mRNAs containing at least one exon-junction complex (EJC) via its interaction with UPF1, promoting the interaction between UPF1 and UPF2. The CBC complex is also involved in 'failsafe' NMD, which is independent of the EJC complex, while it does not participate in Staufen-mediated mRNA decay (SMD). During cell proliferation, the CBC complex is also involved in microRNAs (miRNAs) biogenesis via its interaction with SRRT/ARS2 and is required for miRNA-mediated RNA interference. The CBC complex also acts as a negative regulator of PARN, thereby acting as an inhibitor of mRNA deadenylation. In the CBC complex, NCBP1/CBP80 does not bind directly capped RNAs (m7GpppG-capped RNA) but is required to stabilize the movement of the N-terminal loop of NCBP2/CBP20 and lock the CBC into a high affinity cap-binding state with the cap structure.

  • Sequence similarities

    Belongs to the NCBP1 family.
    Contains 1 MIF4G domain.

  • Cellular localization

    Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 80 kDa nuclear cap-binding protein antibody
    • CBP80 antibody
    • MGC2087 antibody
    • NCBP 80 kDa subunit antibody
    • NCBP antibody
    • ncbp1 antibody
    • NCBP1_HUMAN antibody
    • nuclear cap binding protein subunit 1 antibody
    • nuclear cap binding protein subunit 1, 80kDa antibody
    • Nuclear cap-binding protein subunit 1 antibody
    • Sto1 antibody
    see all

Images

  • Western blot - Anti-CBP80 antibody (ab42389)
    Western blot - Anti-CBP80 antibody (ab42389)
    All lanes : Anti-CBP80 antibody (ab42389) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : Jurkat nuclear extract lysate (ab14844)
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : Hep G2 nuclear extract lysate (ab14660)
    Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 92 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    ab42389 detects a band at 80 kDa which corresponds to the NCBP 80 kDa subunit.

  • Immunocytochemistry/ Immunofluorescence - Anti-CBP80 antibody (ab42389)
    Immunocytochemistry/ Immunofluorescence - Anti-CBP80 antibody (ab42389)
    ICC/IF image of ab42389 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42389, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Immunoprecipitation - Anti-CBP80 antibody (ab42389)
    Immunoprecipitation - Anti-CBP80 antibody (ab42389)

    CBP80 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to CBP80 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 ab42389.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 80kDa; CBP80

References (5)

Publishing research using ab42389? Please let us know so that we can cite the reference in this datasheet.

ab42389 has been referenced in 5 publications.

  • Viphakone N  et al. Co-transcriptional Loading of RNA Export Factors Shapes the Human Transcriptome. Mol Cell 75:310-323.e8 (2019). PubMed: 31104896
  • Chuang TW  et al. A Point Mutation in the Exon Junction Complex Factor Y14 Disrupts Its Function in mRNA Cap Binding and Translation Enhancement. J Biol Chem 291:8565-74 (2016). PubMed: 26887951
  • Schraivogel D  et al. Importin-ß facilitates nuclear import of human GW proteins and balances cytoplasmic gene silencing protein levels. Nucleic Acids Res 43:7447-61 (2015). PubMed: 26170235
  • Massimelli MJ  et al. Interplay between polyadenylate-binding protein 1 and Kaposi's sarcoma-associated herpesvirus ORF57 in accumulation of polyadenylated nuclear RNA, a viral long noncoding RNA. J Virol 87:243-56 (2013). PubMed: 23077296
  • Haedicke J  et al. The brain-specific factor FEZ1 is a determinant of neuronal susceptibility to HIV-1 infection. Proc Natl Acad Sci U S A 106:14040-5 (2009). WB ; Human . PubMed: 19667186

Images

  • Western blot - Anti-CBP80 antibody (ab42389)
    Western blot - Anti-CBP80 antibody (ab42389)
    All lanes : Anti-CBP80 antibody (ab42389) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : Jurkat nuclear extract lysate (ab14844)
    Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 6 : Hep G2 nuclear extract lysate (ab14660)
    Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 92 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    ab42389 detects a band at 80 kDa which corresponds to the NCBP 80 kDa subunit.
  • Immunocytochemistry/ Immunofluorescence - Anti-CBP80 antibody (ab42389)
    Immunocytochemistry/ Immunofluorescence - Anti-CBP80 antibody (ab42389)
    ICC/IF image of ab42389 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab42389, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • Immunoprecipitation - Anti-CBP80 antibody (ab42389)
    Immunoprecipitation - Anti-CBP80 antibody (ab42389)

    CBP80 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to CBP80 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 ab42389.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 80kDa; CBP80

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