Recombinant 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
Related conjugates and formulations
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
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, 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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB |
1/1000. Detects a band of approximately 27-29 kDa (predicted molecular weight: 22 kDa).
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IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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Notes |
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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
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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
modificationsN-glycosylated. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 29851 Human
- Entrez Gene: 54167 Mouse
- Entrez Gene: 64545 Rat
- Omim: 604558 Human
- SwissProt: Q9Y6W8 Human
- SwissProt: Q9WVS0 Mouse
- SwissProt: Q9R1T7 Rat
- Unigene: 56247 Human
see all -
Alternative names
- Activation inducible lymphocyte immunomediatory molecule antibody
- Activation-inducible lymphocyte immunomediatory molecule antibody
- AILIM antibody
see all
Images
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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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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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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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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).
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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).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab224644 has been referenced in 3 publications.
- Huo X et al. Identification of Novel Immunologic Checkpoint Gene Prognostic Markers for Ovarian Cancer. J Oncol 2022:8570882 (2022). PubMed: 36157232
- Sun S et al. Characterization and structural basis of a lethal mouse-adapted SARS-CoV-2. Nat Commun 12:5654 (2021). PubMed: 34580297
- 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 . PubMed: 29614981