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Analysis of Jak3 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0092-LGZ)

Introduction

Official Full Name: Janus kinase 3
Also known as: fae; wil
Enables protein tyrosine kinase activity. Involved in several processes, including negative regulation of FasL production; negative regulation of cytokine production; and regulation of apoptotic process. Acts upstream of or within several processes, including cell surface receptor signaling pathway; peptidyl-tyrosine phosphorylation; and positive regulation of transcription by RNA polymerase II. Predicted to be located in cytoplasm. Is expressed in several structures, including alimentary system; brain; genitourinary system; hemolymphoid system gland; and liver and biliary system. Used to study severe combined immunodeficiency, autosomal recessive, T cell-negative, B cell-positive, Nk cell-positive. Human ortholog(s) of this gene implicated in T-cell acute lymphoblastic leukemia; acute megakaryocytic leukemia; adult T-cell leukemia; and juvenile myelomonocytic leukemia. Orthologous to human JAK3 (Janus kinase 3).




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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