Anti-ADAM10 Antibody

ADAM10 KO Validation in MEF Cells
Loading: 10 μg of lysate Antibodies: ADAM10 A00566-2, 1 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. A00566-2 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

ADAM10 KO Validation in 293 Cells
Loading: 15 μg of lysate Antibodies: ADAM10 A00566-2, 2 μg/mL and beta-actin 3779-1301, 1μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. A00566-2 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines
Loading: 15 μg of lysates per lane. Antibodies: ADAM10 A00566-2, 0.5 μg/mL, ADAM10 24-024, 1 μg/mL, and -actin 3779, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

WB Validation in Human Cell Lines
Loading: 15 μg of lysate Antibodies: ADAM10 A00566-2, 1 μg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. A00566-2 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Western Blot Validation in Mouse Tissues
Loading: 15 μg of lysates per lane. Antibodies: ADAM10 A00566-2, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. A00566-2 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Western Blot Validation in Rat Tissues
Loading: 15 μg of lysates per lane. Antibodies: ADAM10 A00566-2, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. A00566-2 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Immunofluorescence Validation of ADAM10 in MOLT4 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed MOLT4 cells labeling ADAM10 with A00566-2 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunofluorescence Validation of ADAM10 in Mouse Testis
Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse testis labeling ADAM10 with A00566-2 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunofluorescence Validation of ADAM10 in Rat Testis
Immunofluorescent analysis of 4% paraformaldehyde-fixed rat testis labeling ADAM10 with A00566-2 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunofluorescence Validation of ADAM10 in Rat Thymus
Immunofluorescent analysis of 4% paraformaldehyde-fixed rat thymus labeling ADAM10 with A00566-2 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunohistochemistry Validation of ADAM10 in Human Testis
Immunohistochemical analysis of paraffin-embedded human testis tissue using anti-ADAM10 antibody (A00566-2) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of ADAM10 in Mouse Thymus
Immunohistochemical analysis of paraffin-embedded mouse thymus tissue using anti-ADAM10 antibody (A00566-2) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of ADAM10 in primary cultures of human cerebral vascular smooth muscle cells (HC-VSMCs) (Coma et al., 2008)
Detection of ADAM10 expression by anti-ADAM10 antibodies in HC-VSMC cells under control condition or in the presence of 10μM H2O2 (oxidative stress condition) for 6 h. ADAM10 expression was not affected when exposed to oxidative stress.

Regulated Expression Validation of ADAM10 in Human neuroblastoma (SH-SY5Y) cells (Zimmermann et al., 2004)
Protein expression of ADAM10 detected by anti-ADAM10 CT antibodies in control or donepezil treated SH-SY5Y cells. When treated with donepezil, the expression of mature form of ADAM10 (68kD) was up-regulated in membrane compartment as compared to the down-regulation in intracellular fractions, and was not affected in whole cell homogenate.

KD Validation of ADAM10 in Human embryonic kidney 293 cells overexpressing the human APP 695 isoform (HEK/APP) (Gatta et al., 2009)
Western blot analysis of ADAM10 silencing using anti-ADAM10 antibodies in HEK/APP cells. Silencing with ADAM10 siRNA (Ad) significantly decreased ADAM10 expression, and so did with Ferrochelatase siRNA (F) and N-methylprotoporphyrin IX siRNA (N), 67% and 50% reduction respectively