Basic Information
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Description
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- EPHA2 is a receptor tyrosine kinase that mainly causes rejection and adhesion reactions between cells.
- The exons 1-8 of mouse Epha2 gene that encode signal peptide, and extracellular domain are replaced by human counterparts in B-hEPHA2 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human EPHA2 expression is driven by endogenous mouse Epha2 promoter, while mouse Epha2 gene transcription and translation will be disrupted.
- Mouse Epha2 mRNA was detectable in wild-type C57BL/6JNifdc mice. Human EPHA2 mRNA was exclusively detectable in homozygous B-hEPHA2 mice.
- EPHA2 protein was detected in lung, kidney, colon, stomach, and esophagus of wild-type C57BL/6JNifdc mice and homozygous B-hEPHA2 mice, as the antibody can cross-recognize both human and mouse EPHA2.
- Application: This product is used for pharmacodynamics and safety evaluation of drugs.
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Targeting strategy
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Gene targeting strategy for B-hEPHA2 mice. The exons 1-8 of mouse Epha2 gene that encode signal peptide, and extracellular domain are replaced by human counterparts in B-hEPHA2 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human EPHA2 expression is driven by endogenous mouse Epha2 promoter, while mouse Epha2 gene transcription and translation will be disrupted.
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mRNA expression analysis
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Strain specific analysis of EPHA2 mRNA expression in wild-type C57BL/6JNifdc mice and B-hEPHA2 mice by RT-PCR. Lung RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hEPHA2 mice (H/H). Mouse Epha2 mRNA was only detectable in wild-type C57BL/6JNifdc mice, and human EPHA2 mRNA was exclusively detectable in homozygous B-hEPHA2 mice but not in wild-type C57BL/6JNifdc mice.
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Protein expression analysis
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Strain specific EPHA2 expression analysis in homozygous B-hEPHA2 mice by western blot. Lung, kidney, colon, stomach, and esophagus were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hEPHA2 mice (H/H), and analyzed by western blot with anti-EPHA2 antibody (Abcam, ab273118). 40 μg total proteins were loaded for western blotting analysis. EPHA2 was detectable in these tissues of wild-type C57BL/6JNifdc mice and homozygous B-hEPHA2 mice.
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Protein expression analysis in neutrophils
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CD300E expression analysis in F1 B-hCD300E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and F1 B-hCD300E mice, and analyzed by flow cytometry with anti-CD300E antibody (BD Biosciences, 566315). hCD300E was not detectable in neutrophils.
