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Analysis of CD177 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0086-LGZ)

Introduction

Official Full Name: CD177 molecule<br />Also known as: NB1; PRV1; HNA2A; PRV-1; HNA-2a; NB1 GP<br />This gene encodes a glycosylphosphatidylinositol (GPI)-linked cell surface glycoprotein that plays a role in neutrophil activation. This protein binds platelet endothelial cell adhesion molecule-1 and is involved in neutrophil transport. Mutations in this gene are associated with myeloproliferative disorders. This gene is overexpressed in patients with polycythemia vera. Autoantibodies against this protein can cause pulmonary transfusion reactions and may be associated with Wegener's granulomatosis. A related pseudogene adjacent to this gene on chromosome 19 has been identified.




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