Ex Vivo Immune Response Assays - Th1, Th2, and Th17 Responses
CONTACT CUSTOMER SERVICE
ADD TO FAVORITES
ADD TO PANEL
OVERVIEW
DOCUMENTS/REFERENCES
BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
Ex vivo immune response assays are a suite of assays that can be used and also customized to evaluate the effects of a test article on the immune responses. Measurements of cytokines in cell culture supernatants, cell proliferation, or Th1, Th2, and Th17 cell responses are widely used to study immune responses. We offer a variety of ex vivo assays for evaluating the efficacy of test articles on these immune responses.
ORGANISM
Mouse
ASSAY INFORMATION
ASSAY TYPE
Functional
TESTING INFORMATION
PROCEDURE SUMMARY
Spleens or lymph nodes are harvested from mice and crushed through 70-µm strainer with the plunger of 3 mL syringe to obtain a single cell suspension. The red blood cells are lysed by ACK buffer. After washing, cells are adjusted maximally to 2×106 cells/mL. For detection of Th1, Th2 and Th17 cells, the cultured single cells are activated with Phorbol 12-myristate 13-acetate (PMA, 50 ng/mL) / ionomycin (500 ng/mL) or other antigens in the RPMI medium for 4 hours at 37°C in a 5% CO2 atmosphere. Other culture conditions can also be requested. The cells are stained with fluorescence-conjugated antibodies for markers, including CD3, CD4, IFN-γ, IL-4, and IL-17 according to the manufacturer’s protocols for flow cytometry analysis. We define Th1 as CD3+CD4+IFN-γ+ cells, Th2 as CD3+CD4+IL-4+ cells, and Th17 cells as CD3+CD4+IL-17+ cells. All antibodies are purchased commercially and tested for optimal dilution in-house. Fluorescence is acquired and analyzed using a flow cytometer system. The results are expressed as the numbers of specific cell populations as well as the percentage of positive cells.
CRITERIA FOR SIGNIFICANCE
ANOVA and Dunnett's test are used to ascertain the difference between vehicle control and treated mice. Significance is set at p<0.05 level.
TURNAROUND TIME
STANDARD
15 Days
CLINICAL RELEVANCE
THERAPEUTIC AREA
Inflammation/Allergy
ADDITIONAL INFORMATION
BRAND
PDS
TESTING LOCATION
Taiwan - Taipei
OTHER INFORMATION
We will readily accommodate client-specified alterations. These studies are performed at our AAALAC accredited BSL-2 laboratory in Taipei, Taiwan. All aspects of this work are performed in general accordance with the Guide for the Care and Use of laboratory animals (National Academy Press, Washington, DC, 2011). The study protocol was approved by the Pharmacology Discovery Services IACUC and is performed with the oversight of veterinarians to assure the humane treatment of laboratory animals.
ITEM DOES NOT EXIST
© Copyright 2026, Pharmacology Discovery Services – Privacy Policy | Cookie Notice | Cookie Preferences