Anti-ICOS antibody [EPR20560] (ab224644)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20560] to ICOS
- Suitable for: WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-ICOS antibody [EPR20560]
See all ICOS primary antibodies -
Description
Rabbit monoclonal [EPR20560] to ICOS -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P MouseRatHumanIP HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20560 -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICOS antibody [EPR20560] (ab224644)
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).
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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 secondBlocking and dilution buffer: 5% NFDM/TBST
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICOS antibody [EPR20560] (ab224644)
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).
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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 secondsBlocking and dilution buffer: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID: 11169414).
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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 secondsBlocking 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 ).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICOS antibody [EPR20560] (ab224644)
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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICOS antibody [EPR20560] (ab224644)
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).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICOS antibody [EPR20560] (ab224644)
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).
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