Overview

  • Product nameAnti-ROC1 antibody
    See all ROC1 primary antibodies
  • Description
    Rabbit polyclonal to ROC1
  • Tested applicationsSuitable for: IHC-P, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Caenorhabditis elegans, Drosophila melanogaster, Non Human Primates, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human ROC1.

    (Peptide available as ab97983.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; Jurkat; HEK293; MCF7.

Properties

Applications

Our Abpromise guarantee covers the use of ab86862 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 10 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 12 kDa).

Target

  • FunctionE3 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 specificityWidely expressed.
  • PathwayProtein modification; protein ubiquitination.
  • Sequence similaritiesBelongs to the RING-box family.
    Contains 1 RING-type zinc finger.
  • DomainThe RING-type zinc finger domain is essential for ubiquitin ligase activity. It coordinates an additional third zinc ion.
  • Cellular localizationCytoplasm. 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
    • ZYP protein antibody
    see all

Anti-ROC1 antibody images

  • All lanes : Anti-ROC1 antibody (ab86862) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 12 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 8 minutes
  • ICC/IF image of ab86862 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86862, 10µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 10µg/ml.
  • IHC image of ROC1 staining in human normal kidney 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 ab86862, 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.
  • Anti-ROC1 antibody (ab86862) at 1 µg/ml + ROC1 protein (ab75321) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 90 seconds

References for Anti-ROC1 antibody (ab86862)

ab86862 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Loading amount 40 µg
Specification Mouse embryonic fibroblasts
Gel Running Conditions Reduced Denaturing (4-20% Tris-Gly gel)
Blocking step LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
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Submitted Jan 13 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HeLa cells)
Total protein in input 200 µg
Specification HeLa cells
Immuno-precipitation step Protein A/G
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Submitted Apr 06 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HCT116 cells)
Loading amount 30 µg
Specification HCT116 cells
Gel Running Conditions Reduced Denaturing (4-20% Tris-Gly gel)
Blocking step LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jan 13 2011

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