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Analysis of TNFRSF8 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0070-LGZ)

Introduction

Official Full Name: TNF receptor superfamily member 8
Also known as: CD30; Ki-1; D1S166E
The protein encoded by this gene is a member of the TNF receptor superfamily. This receptor is expressed on activated T cells and B cells, but not on resting T cells and B cells. TRAF2 and TRAF5 can interact with this receptor and mediate signal transduction leading to NF-kappaB activation. This receptor is a positive regulator of apoptosis and has also been shown to limit the proliferative potential of autoreactive CD8 effector T cells and protect the body from autoimmunity. Two alternatively spliced transcript variants encoding different isoforms 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
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