T Cell-based Intracellular Staining Assay

Intracellular Staining Assay for Cancer Epitope Analysis

The intracellular staining approach is a prevailing method used to quantitatively measure the immune response-related proteins produced in cells, including cytokines and granzyme, and perforin, to assess cell responses to various stimuli. With a deep understanding of T cell immunity and successful experience in T cell phenotype and function study, Creative Biolabs offers T cell-based intracellular staining assay to study cancer epitope-induced specific T cell cytokine, granzyme, and perforin production, providing scientific information on the determination of targets for cancer immunotherapy.

Fig.1 Flow cytometric detection of cytokines in biological samples.¹

Intracellular Staining Assay Services at Creative Biolabs

Intracellular staining is a versatile and flexible assay to monitor cytokines, granzyme, and perforin production and accumulation in cell samples. When combined with the powerful flow cytometry technique, the assay is functional to assess antigen-specific T cell response and identify immunogenic cancer epitopes.

Workflow

Cell Preparation

Crude samples, such as cell mixtures or clinical human tissues, can be used for evaluation.
Freshly dissociated cells permit convenient determination of T cell activation status in vivo.
PBMC or T cells are activated by mitogens or antigens in vitro to mimic various stimulation conditions in vivo.

Epitope-specific Intracellular Molecules Identification

We provide a comprehensive quantification service on epitope-specific intracellular molecules using intracellular staining in combination with ELISPOT assay.

List of Cytokines and Effectors Assessed by Our Intracellular Staining Assay
IL-6	IL-2	IL-3	GM-CSF	TNFa	TNF	IL-15
IL-12	IL-8	IL-5	IL-1b	IL-13	IL-17	IFNg
Perforin	Granzyme A	Granzyme B	CCL4/MIP-1b	CCL3/MIP-1a

Comparison of ELISPOT Assay and Intracellular Staining Assay

ELISPOT Assay

Analysis pools of cancer epitope peptides
Recommended for the first round of large-scale screening
A small number of cells are needed

Intracellular Staining Assay

Monitor the T-cell response to a specific peptide or a small number of peptides
Providing comprehensive information on phenotype and function
Cost larger numbers of cells

Features of T Cell-based Intracellular Staining Assay

Effector protein molecules are limited to the rough endoplasmic reticulum to facilitate detection.
Crude samples can be directly used for tests.
Detect intra- and extra-cellular protein markers in every single target cell simultaneously.

Key Applications

Determine cytokines, granzyme A, granzyme B, and perforin production;
Evaluate the proportion and numbers of epitope-specific T cells;
Analysis of phenotype parameters of the corresponding cell mixture.

Creative Biolabs offers comprehensive assays to provide reliable and accurate data on cytokines and cytotoxic molecule production in activated T cells. Besides intracellular staining, we also provide the following assays based on the measurement of cytokines. Creative Biolabs is devoted to providing the most proper assays to fit your needs and ensuring high-quality reports. Please feel free to contact us to discuss your projects.

Reference

Sachdeva, N.; Asthana, D. Cytokine quantitation: technologies and applications. Frontiers in bioscience: a journal and virtual library. 2007,12: 4682–4695.

For Research Use Only | Not For Clinical Use