Mouse ICOS Antibody

R&D Systems | Catalog # AF168

R&D Systems

Key Product Details

Species Reactivity

Mouse

Applications

Western Blot, Blockade of Receptor-ligand Interaction

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all ICOS Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant mouse ICOS
Met1-Leu142
Accession # CAB71153

Specificity

Detects mouse ICOS in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross‑reactivity with recombinant mouse (rm) CTLA-4, rmCD28, and rmPD-1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse ICOS Antibody

Detection of ICOS by Flow Cytometry

Detection of ICOS by Flow Cytometry

ICOS transmembrane domain-mediated helper functions. a, b Representative contour plots and frequencies of IgMa+Fashi GC B cells among the total CD19+ cells (a) and CXCR5hiPD-1hi TFH cells among the transferred OT-II cells (b). Each symbol in scatter plots represents one mouse, and the lines denote the means. The data presented were pooled from 3 independent experiments (**P < 0.01 and **** P < 0.0001, as determined by Mann–Whitney tests). c Surface CAR staining of Jurkat cells lentivirally transduced to express CD19-reactive BBz CAR, which has the original CD8 transmembrane domain, or ICOSTM-BBz CAR, which has the ICOS transmembrane domain. d Representative FACS profiles (left; the numbers indicate the event percentages in the gates) and summarized frequencies of CAR-expressing Jurkat cells in conjugation with TAMARA-labeled CD19+ Farage B cells (right). Each line connecting two dots represents one independent experiment (****P < 0.0001, as determined by paired t-test) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30523347), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICOS by Flow Cytometry

Detection of ICOS by Flow Cytometry

The ICOS transmembrane domain, but not the cytoplasmic domain, mediates the costimulation of calcium mobilization. a Surface ICOS staining of Jurkat cells lentivirally transduced with an empty vector or vectors expressing full-length ICOS or ICOS-NT. b Example of gating GFP+ stably transduced Jurkat cells and GFP− spike-in nontransduced control Jurkat cells for the calcium mobilization assay. c Calcium mobilization in Jurkat cells of the indicated types and the respective internal GFP- control cells after stimulation with biotinylated anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. d Surface ICOS staining of Jurkat cells lentivirally transduced with vectors expressing mutant ICOS molecules of the indicated types. e Surface ICOS staining after normalization of expression between ICOS-NT and ICOS-CD44TM-NT. f Calcium mobilization in Jurkat cells of the indicated types conducted as for c. The data represent the results of three independent experiments Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30523347), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICOS by Flow Cytometry

Detection of ICOS by Flow Cytometry

ICOS transmembrane domain-mediated helper functions. a, b Representative contour plots and frequencies of IgMa+Fashi GC B cells among the total CD19+ cells (a) and CXCR5hiPD-1hi TFH cells among the transferred OT-II cells (b). Each symbol in scatter plots represents one mouse, and the lines denote the means. The data presented were pooled from 3 independent experiments (**P < 0.01 and **** P < 0.0001, as determined by Mann–Whitney tests). c Surface CAR staining of Jurkat cells lentivirally transduced to express CD19-reactive BBz CAR, which has the original CD8 transmembrane domain, or ICOSTM-BBz CAR, which has the ICOS transmembrane domain. d Representative FACS profiles (left; the numbers indicate the event percentages in the gates) and summarized frequencies of CAR-expressing Jurkat cells in conjugation with TAMARA-labeled CD19+ Farage B cells (right). Each line connecting two dots represents one independent experiment (****P < 0.0001, as determined by paired t-test) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30523347), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICOS by Flow Cytometry

Detection of ICOS by Flow Cytometry

The ICOS transmembrane domain mediates calcium costimulation and is required for p85 recruitment. a Surface ICOS staining of Jurkat cells stably expressing ICOS, ICOS-CD44TM, or ICOS-NT molecules. b Calcium mobilization in Jurkat cells expressing ICOS molecules of the indicated types after stimulation with biotinylated anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. c Immunoblotting of phospho-ZAP70 and phospho-PLC gamma in Jurkat cells expressing the indicated ICOS molecules 30 s after stimulation with anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. d Input (3% total lysates) or anti-ICOS immunoprecipitates of the indicated Jurkat cells before or 1 minute after ICOS crosslinking stimulation were probed by Western blotting for p85 and ICOS. e Immunoblotting of the indicated molecules in ICOS or ICOS-CD44TM Jurkat cells following anti-ICOS crosslinking stimulation for the indicated periods of time. Left, representative pAkt patterns; right, normalized pAkt levels of three independent experiments Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30523347), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ICOS by Flow Cytometry

Detection of ICOS by Flow Cytometry

The ICOS transmembrane domain mediates calcium costimulation and is required for p85 recruitment. a Surface ICOS staining of Jurkat cells stably expressing ICOS, ICOS-CD44TM, or ICOS-NT molecules. b Calcium mobilization in Jurkat cells expressing ICOS molecules of the indicated types after stimulation with biotinylated anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. c Immunoblotting of phospho-ZAP70 and phospho-PLC gamma in Jurkat cells expressing the indicated ICOS molecules 30 s after stimulation with anti-CD3 (0.2 μg/ml) and anti-ICOS (2 μg/ml) antibodies coligated by streptavidin. The data represent the results of three independent experiments. d Input (3% total lysates) or anti-ICOS immunoprecipitates of the indicated Jurkat cells before or 1 minute after ICOS crosslinking stimulation were probed by Western blotting for p85 and ICOS. e Immunoblotting of the indicated molecules in ICOS or ICOS-CD44TM Jurkat cells following anti-ICOS crosslinking stimulation for the indicated periods of time. Left, representative pAkt patterns; right, normalized pAkt levels of three independent experiments Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30523347), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse ICOS Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 10-40 µg/mL of this antibody will block 50% of the binding of 500 ng/mL of Biotinylated Recombinant Human B7-H2 Fc Chimera to immobilized Recombinant Mouse ICOS (Catalog # 168-CS) coated at 1 µg/mL (100 µL/well). At 75 μg/mL, this antibody will block >90% of the binding.

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse ICOS Fc Chimera (Catalog # 168-CS)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ICOS

Mouse ICOS (inducible co-stimulator), also called AILIM (activiation-inducible lymphocyte immunomediatory molecule) and CRP-1 (CD28-related protein-1), is a member of the CD28 family of immune costimulatory receptors. Other family members are CD28, CTLA-4 and PD-1. Mouse ICOS is a homodimeric type I transmembrane protein consisting of 200 amino acids (aa) with a putative 20 aa signal sequence, a 122 aa extracellular domain, a 23 aa transmembrane region, and a 35 aa cytoplasmic domain. ICOS shares approximately 39% aa similarity with CD 28 and CTLA-4. Mouse and human ICOS share approximately 72% aa identity. ICOS is expressed on most CD45RO+ cells. ICOS expression is upregulated within approximately 24‑48 hours of activation on Th primed cells. B7-H2, a member of the B7 family of costimulatory ligands, has been identified as the ICOS ligand. The B7-H2/ICOS interaction appears to play roles in T cell dependent B cell activation and Th differentiation.

References

  1. Coyle, A.J. and J.C. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.
  2. Mages, H.W. et al. (2000) Eur. J. Immunol. 30:1040.
  3. Yoshinaga, S.K. et al. (1999) Nature 402:827.
  4. Hutloff, A. et al. (1999) Nature 397:263.
  5. Aicher, A. et al. (2000) J. Immunol. 164:4689.
  6. Coyle, A.J. et al. (2000) Immunity 13:95.
  7. Gonzalo, J.A. et al. (2001) J. Immunol. 166:1.

Long Name

Inducible Co-stimulator

Alternate Names

AILIM, CD278, CRP-1

Entrez Gene IDs

29851 (Human); 54167 (Mouse); 102116512 (Cynomolgus Monkey)

Gene Symbol

ICOS

UniProt

CAB71153

Additional ICOS Products

Product Documents for Mouse ICOS Antibody

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Product Specific Notices for Mouse ICOS Antibody

For research use only

Citations for Mouse ICOS Antibody

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