Analysis of ALK Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0009-LGZ)

Introduction

Official Full Name: ALK receptor tyrosine kinase<br />Also known as: ALK1; CD246; NBLST3<br />This gene encodes a receptor tyrosine kinase that belongs to the insulin receptor superfamily. The protein consists of an extracellular domain, a hydrophobic stretch corresponding to a single-pass transmembrane region, and an intracellular kinase domain. It plays an important role in the development of the brain and affects specific neurons in the nervous system. The gene has been found to be rearranged, mutated or amplified in a range of tumors, including anaplastic large cell lymphoma, neuroblastoma and non-small cell lung cancer. The chromosomal rearrangements are the most common genetic alterations in this gene, which result in creation of multiple fusion genes in tumorurigenesis, including ALK (chromosome 2)/EML4 (chromosome 2), ALK/RANBP2 (chromosome 2), ALK/ATIC (chromosomal some 2), ALK/TFG (chromosome 3), ALK/NPM1 (chromosome 5), ALK/SQSTM1 (chromosome 5), ALK/KIF5B (chromosome 10), ALK/CLTC (chromosome 17), ALK/TPM4 (chromosome 19) , and ALK/MSN (chromosome X).

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Principle Applications Procedure Materials Notes Related Products

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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