Overview

  • Product name

    Anti-Ubiquitin antibody [6C11-B3]
    See all Ubiquitin primary antibodies
  • Description

    Mouse monoclonal [6C11-B3] to Ubiquitin
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Cow Ubiquitin conjugated to keyhole limpet haemocyanin.
    Database link: P0CG53

  • Positive control

    • ICC/IF: HeLa cells. WB: Mixture of human cell lines, cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab223368 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
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 10 kDa (predicted molecular weight: 34 kDa).

Target

  • Relevance

    Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.
  • Cellular localization

    Cell Membrane, Cytoplasmic and Nuclear
  • Database links

  • Alternative names

    • Epididymis secretory protein Li 50 antibody
    • FLJ25987 antibody
    • HEL S 50 antibody
    • MGC8385 antibody
    • Polyubiquitin B antibody
    • RPS 27A antibody
    • RPS27A antibody
    • UBA 52 antibody
    • UBA 80 antibody
    • UBA52 antibody
    • UBA80 antibody
    • UBB antibody
    • UBB_HUMAN antibody
    • UBC antibody
    • UBCEP 1 antibody
    • UBCEP 2 antibody
    • UBCEP1 antibody
    • UBCEP2 antibody
    • Ubiquitin antibody
    • Ubiquitin B antibody
    see all

Images

  • 2% formaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Ubiquitin (green) using ab223368 at 1/100 dilution in ICC/IF.

    Secondary: Goat Anti-Mouse FITC at 1/200 dilution for 2 hours at RT.

    Counterstain: DAPI (blue) nuclear stain at 1/40000 for 2 hours RT.

    (A) DAPI nuclear stain (B) ab223368 (C) Merge.

  • Anti-Ubiquitin antibody [6C11-B3] (ab223368) at 1/1000 dilution + Human cell line mixture, cell lysate at 15 µg

    Secondary
    Sheep Anti-Mouse IgG HRP

    Predicted band size: 34 kDa
    Observed band size: 10 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 1.5% BSA.

    Primary antibody was incubated for 2 hours, the secondary for 1 hour at room temperature.

    The cell line mixture contains: A431 (human epidermoid carcinoma cell line), A549 (human lung carcinoma cell line), HCT116 (human colon cancer), HeLa (human epithelial cell line from cervix adenocarcinoma), HEK-293 (human epithelial cell line from embryonic kidney), HepG2 (human liver hepatocellular carcinoma cell line), HL-60 (human promyelocytic leukemia cell line), HUVEC (human umbilical vein endothelial cell line), Jurkat (human T cell leukemia cell line from peripheral blood), MCF7 (human breast adenocarcinoma cell line), PC-3 (human prostate adenocarcinoma cell line) and T98G (human brain glioblastoma).

  • 2% formaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Ubiquitin (red) using ab223368 at 1/100 dilution in ICC/IF.

    Secondary: Goat Anti-Mouse APC at 1/200 dilution for 2 hours at RT.

    Counterstain: DAPI (blue) nuclear stain at 1/40000 for 2 hours RT.

    (A) DAPI nuclear stain (B) ab223368 (C) Merge.

References

ab223368 has not yet been referenced specifically in any publications.

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