T Cell-based RNA/DNA Detection Assay

Assessing Cytokine Production by PCR

Detection and analysis of cytokine production are principal in understanding the host response to multiple antigenic stimuli. Cytokines are typically measured using ELISA, cytometric bead array, and intracellular staining methods with cell cultures or supernatants. However, in some circumstances, only a small number of cells are available, or tissue samples are applied to detect cytokines. The measurement of the mRNA expression level of cytokines is a useful approach to estimating cytokine production potency.

The PCR-based molecular technique--reverse transcription-polymerase chain reaction (RT-PCR) is adopted to amplify target genes using an mRNA template. The RT-PCR method is also called quantitative fluorescent RT-PCR and is of high sensitivity and specificity for detecting 6 DNA and 40 RNA copies/reaction, respectively.

Fig.1 The process of RT-PCR.¹

T Cell-based RNA/DNA Detection Assay Services at Creative Biolabs

Creative Biolabs provides a practical method to determine cytokine gene expression using the RT-PCR technique to help clients profile cancer epitopes by detecting T cell activation-dependent cytokine production. Cytokines present in various samples can be measured by determining cytokine mRNA abundance at the transcription level, as the transcription process primarily regulates cytokine production.

Workflow

Sample Source

Range of Detection

The RNA/DNA detection assay can be used to detect any cytokine of interest.

Advantages of the Assay

• Multiple genes and sequences can be amplified from a single RNA.
• Primers for the amplification of a specific region of the gene are designed and evaluated for specificity and fitness for PCR procedures before the formal experiment.
• The amplification procedure is optimized for an individual experiment.
• The results can calculate using absolute quantification or relative quantification method.

Creative Biolabs is a reliable partner for cancer epitope discovery. We have assembled a team of experts in tumor immunology and a robust PCR technical platform to provide accurate, sensitive, and specific cytokine detection services using the classical PCR method. Interested in this specific and efficient assay? Please contact us and deliver your requirements.

References

1. Roberts, T. C.; et al. Detection and quantification of extracellular microRNAs in murine biofluids. Biological procedures online. 2014, 16(1): 5.

For Research Use Only | Not For Clinical Use