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Analysis of IL15 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0047-LGZ)

Introduction

Official Full Name: interleukin 15<br />Also known as: IL-15<br />The protein encoded by this gene is a cytokine that regulates the activation and proliferation of T cells and natural killer cells. This cytokine shares many biological activities with interleukin-2. They were found to bind a common hematopoietin receptor subunit and may compete for the same receptor, thereby negatively regulating each other's activity. The number of CD8+ memory cells was shown to be controlled by the balance between this cytokine and IL-2. This cytokine induces the activation of JAK kinases, as well as the phosphorylation and activation of the transcriptional activators STAT3, STAT5 and STAT6. Studies of the mouse counterpart indicated that this cytokine may prevent apoptosis by increasing the expression of the apoptosis inhibitor BCL2L1/BCL-x(L) through the transcriptional activation activity of STAT6. Alternative splicing transcript variants of this gene have been reported.




Principle

Under certain conditions, two single strands of nucleic acid with certain homology can be specifically hybridized to form double strands according to the principle of base complementarity. Generally, DNA molecules to be detected are digested with restriction enzymes, separated by agar-gel electrophoresis, denatured and transferred to nitrocellulocellulose film or nylon film or other solid phase support according to their position in the gel, fixed and then reacted with DNA probes labeled with isotopes or other markers. This is followed by autoradiography or an enzyme reaction to detect the amount of specific DNA molecules. If the object to be tested contains a sequence that is complementary to the probe, the two are combined by the principle of base complementarity, and the free probe is washed and detected by self-development or other suitable techniques, thus revealing the fragment to be tested and its relative size.

Applications

Gene Rearrangement Detection

Procedure

1. Sample Processing
2. DNA Extraction and Digestion
3. Gel Electrophoresis
4. Gel Pretreatment
5. Transfer membrane
6. Probe Labeling
7. Prehybridization (blocking)
8. Southern hybridization
9. Membrane washing
10. Autoradiographic Assay
11. Results Analysis

Materials

Sample: DNA, Bacterial Fluid/Tissue/Cell