Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Analysis of ABL1 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0053-LGZ)

Introduction

Official Full Name: ABL proto-oncogene 1, non-receptor tyrosine kinase
Also known as: ABL; JTK7; p150; c-ABL; v-abl; CHDSKM; c-ABL1; BCR-ABL; bcr/abl
This gene is a proto-oncogene that encodes a tyrosine kinase protein involved in a variety of cellular processes including cell division, adhesion, differentiation and stress response. The activity of this protein is negatively regulated by its SH3 domain, thus, deletion of the region encoding this domain results in the generation of an oncogene. This ubiquitously expressed protein has DNA-binding activity regulated by CDC2-mediated phosphorylation, suggesting a cell cycle function. In various leukemias, this gene has been found fused to a variety of translocation partner genes, most notably the t(9;22) translocation, resulting in a fusion to the 5' end of the breakpoint cluster region gene. Alternative splicing of this gene results in two transcript variants containing an alternative first exon spliced to the remaining common exon.




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
Advertisement