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Analysis of DDIT3 Gene Rearrangement by Southern Blot Technology (CAT#: STEM-MHT-0067-LGZ)

Introduction

Official Full Name: DNA damage inducible transcript 3<br />Also known as: CHOP; CEBPZ; CHOP10; CHOP-10; GADD153; AltDDIT3; C/EBPzeta<br />This gene encodes a member of the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors. The protein acts as a dominant-negative inhibitor by forming heterodimers with other C/EBP members such as C/EBP and LAP (liver activator protein), and preventing their DNA-binding activity. This protein is involved in adipogenesis and erythropoiesis, is activated by endoplasmic reticulum stress, and promotes apoptosis. This gene is fused to FUS on chromosome 16 or translocation-induced EWSR1 on chromosome 22 to produce a chimeric protein in myxoid liposarcoma or Ewing's sarcoma. Multiple alternatively spliced transcript variants encoding two isoforms of different lengths have 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