Overview

  • Product name

    Anti-RBX1 antibody [EPR6850(B)]
    See all RBX1 primary antibodies
  • Description

    Rabbit monoclonal [EPR6850(B)] to RBX1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human RBX1 aa 50-150 (C terminal).
    Database link: P62877

  • Positive control

    • Human colon tissue; MCF7, HepG2, HeLa and Jurkat whole cell lysate (ab7899).
  • 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 ab133565 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
Flow Cyt 1/100 - 1/1000.

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

IP 1/10 - 1/100.
ICC/IF 1/100 - 1/250.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/10000. Predicted molecular weight: 12 kDa.

Target

  • Function

    E3 ubiquitin ligase component of multiple cullin-RING-based E3 ubiquitin-protein ligase complexes which mediate the ubiquitination and subsequent proteasomal degradation of target proteins, including proteins involved in cell cycle progression, signal transduction, transcription and transcription-coupled nucleotide excision repair. The functional specificity of the E3 ubiquitin-protein ligase complexes depends on the variable substrate recognition components. As a component of the CSA complex promotes the ubiquitination of ERCC6 resulting in proteasomal degradation. Through the RING-type zinc finger, seems to recruit the E2 ubiquitination enzyme, like CDC34, to the complex and brings it into close proximity to the substrate. Probably also stimulates CDC34 autoubiquitination. May be required for histone H3 and histone H4 ubiquitination in response to ultraviolet and for subsequent DNA repair. Promotes the neddylation of CUL1, CUL2, CUL4 and CUL4 via its interaction with UBE2M.
  • Tissue specificity

    Widely expressed.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Belongs to the RING-box family.
    Contains 1 RING-type zinc finger.
  • Domain

    The RING-type zinc finger domain is essential for ubiquitin ligase activity. It coordinates an additional third zinc ion.
  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • BA554C12.1 antibody
    • E3 ubiquitin-protein ligase RBX1 antibody
    • FLJ60363 antibody
    • MGC13357 antibody
    • MGC1481 antibody
    • OTTHUMP00000028983 antibody
    • Protein ZYP antibody
    • Rbx 1 antibody
    • Rbx1 antibody
    • RBX1_HUMAN antibody
    • Regulator of cullins 1 antibody
    • Ring box 1 antibody
    • Ring box 1 E3 ubiquitin protein ligase antibody
    • RING box protein 1 antibody
    • RING finger protein 75 antibody
    • RING finger protein antibody
    • RING-box protein 1 antibody
    • Ringbox protein 1 antibody
    • RNF 75 antibody
    • RNF75 antibody
    • ROC 1 antibody
    • ROC1 antibody
    • ZYP protein antibody
    see all

Images

  • All lanes : Anti-RBX1 antibody [EPR6850(B)] (ab133565) at 1/1000 dilution

    Lane 1 : MCF7 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 12 kDa

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling RBX1 with ab133565 at 1/100 dilution.

  • Overlay histogram showing HeLa cells stained with ab133565 (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 (ab133565, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • ab133565 at 1/70 immunoprecipitating RBX1 in HepG2 whole cell lysate.

    For western blotting, a primary antibody was used at 1/1000 and a HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:

  • Salehi-Tabar R  et al. The Tumor Suppressor FBW7 and the Vitamin D Receptor Are Mutual Cofactors in Protein Turnover and Transcriptional Regulation. Mol Cancer Res 17:709-719 (2019). Read more (PubMed: 30606768) »
  • Li Y  et al. Heterozygous deletion of chromosome 17p renders prostate cancer vulnerable to inhibition of RNA polymerase II. Nat Commun 9:4394 (2018). Read more (PubMed: 30349055) »
See all 4 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 14 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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