Overview

  • Product name

    Anti-ICOS antibody [EPR20560]
    See all ICOS primary antibodies
  • Description

    Rabbit monoclonal [EPR20560] to ICOS
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ICOS aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9Y6W8

  • Positive control

    • WB: His-tagged human ICOS (aa21-140-GGGGS-aa162-199) recombinant protein; human thymus and tonsil tissue lysate; HuT-78 cell lysate. IP: Human tonsil tissue lysate. IHC-P: Human tonsil, stomach and cervix cancer tissue; mouse spleen tissue; rat spleen tissue.
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20560
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab224644 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 27-29 kDa (predicted molecular weight: 22 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.

Target

  • Function

    Enhances all basic T-cell responses to a foreign antigen, namely proliferation, secretion of lymphokines, up-regulation of molecules that mediate cell-cell interaction, and effective help for antibody secretion by B-cells. Essential both for efficient interaction between T and B-cells and for normal antibody responses to T-cell dependent antigens. Does not up-regulate the production of interleukin-2, but superinduces the synthesis of interleukin-10. Prevents the apoptosis of pre-activated T-cells. Plays a critical role in CD40-mediated class switching of immunoglobin isotypes.
  • Tissue specificity

    Activated T-cells. Highly expressed on tonsillar T-cells, which are closely associated with B-cells in the apical light zone of germinal centers, the site of terminal B-cell maturation. Expressed at lower levels in thymus, lung, lymph node and peripheral blood leukocytes. Expressed in the medulla of fetal and newborn thymus.
  • Involvement in disease

    Immunodeficiency, common variable, 1
  • Sequence similarities

    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Activation inducible lymphocyte immunomediatory molecule antibody
    • Activation-inducible lymphocyte immunomediatory molecule antibody
    • AILIM antibody
    • CD278 antibody
    • CD278 antigen antibody
    • CD28-related Protein-1 antibody
    • CRP1 antibody
    • CVID1 antibody
    • ICOS antibody
    • ICOS_HUMAN antibody
    • Inducible costimulator antibody
    • Inducible T cell co stimulator antibody
    • Inducible T-cell costimulator antibody
    • MGC39850 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling ICOS with ab224644 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and membranous staining in a population of lymphoid cells of human stomach (magnification x200) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution + His-tagged human ICOS (aa21-140-GGGGS-aa162-199) recombinant protein, 10 ng

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

    Developed using the ECL technique.

    Predicted band size: 22 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking and dilution buffer: 5% NFDM/TBST 

  • ICOS was immunoprecipitated from 0.35 mg of human tonsil tissue lysate with ab224644 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224644 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Human tonsil tissue lysate 10 μg (Input).
    Lane 2: ab224644 IP in human tonsil tissue lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224644 in human tonsil tissue lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.


  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling ICOS with ab224644 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and membranous staining in a population of lymphoid cells of human tonsil (PMID: 20207847), (magnification x200) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution

    Lane 1 : Human tymus tissue lysate
    Lane 2 : HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG H&L (HRP))

    Developed using the ECL technique.

    Predicted band size: 22 kDa
    Observed band size: 27-29 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11169414).

  • All lanes : Anti-ICOS antibody [EPR20560] (ab224644) at 1/1000 dilution

    Lane 1 : Untreated human tonsil tissue lysate
    Lane 2 : Human tonsil tissue lysate treated with PNGase F

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG H&L (HRP))

    Developed using the ECL technique.

    Predicted band size: 22 kDa
    Observed band size: 22,27-29 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

    When untreated two distinct bands of 27kDa and 29kDa were observed. Treatment with PNGase F resulted in a single band with a molecular of 22kDa.

    The expression profile observed is consistent with what has been described in the literature (PMID: 11169414 ). 

  • Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling ICOS with ab224644 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and membranous staining in tumor infiltrating lymphoid cells of human cervix cancer (PMID: 23802069) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ICOS with ab224644 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and membranous staining in a population of lymphoid cells of mouse spleen is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling ICOS with ab224644 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and membranous staining in a population of lymphoid cells of rat spleen is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

References

This product has been referenced in:

  • Shibutani M  et al. A comparison of the local immune status between the primary and metastatic tumor in colorectal cancer: a retrospective study. BMC Cancer 18:371 (2018). IHC-P . Read more (PubMed: 29614981) »
See 1 Publication for this product

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