Targeting strategy
Gene targeting strategy for B-hADAM17 mice. The exons 6-15 of the mouse Adam17 gene that encodes the extracellular domain was replaced by human ADAM17 exons 6-15 in B-hADAM17 mice.
Protein expression analysis
Western blot analysis of ADAM17 protein expression in homozygous B-hADAM17 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H), and then analyzed by western blot with anti-ADAM17 antibody. 3 μg total proteins were loaded for simple western analysis. Data analysed by JESS Automated Western Blotting. ADAM17 was detected in spleen, lung, and kidney. The antibody cross recognizes human and mouse ADAM17. Black arrows: mature ADAM17 (mADAM17) and its proform (pADAM17).
mRNA expression analysis
Strain specific analysis of ADAM17 mRNA expression in wild-type C57BL/6 mice and B-hADAM17 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADAM17 primers. Mouse Adam17 mRNA was detectable only in wild-type C57BL/6 mice. Human ADAM17 mRNA was detectable only in homozygous B-hADAM17 mice but not in wild-type mice.
Human ADAM17 antibody binding assay in DCs of spleen
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in B cells of spleen
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in Macrophages of spleen
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in Monocytes of spleen
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in Neutrophils of spleen
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in DCs of liver
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Liver were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in B cells of liver
Human ADAM17 antibody binding assay in Macrophages of liver
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Liver were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
Human ADAM17 antibody binding assay in Epithelial cells of small intestine
Anti-human ADAM17 antibody binding assessment in B-hADAM17 mice. Liver were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAM17 mice (H/H) and analyzed by flow cytometry. Anti-hADM17 antibody hADAM17 Ab-A (provided by a client) can bind B-hADAM17 mice, but not to C57BL/6 mice.
