ADAM10 Antibody

• Catalog Number: orb1239162
• Category: Tools
• Description: ADAM10 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, WB
• Isotype: IgG
• Immunogen: Anti-ADAM10 antibody (orb1239162) was raised against a peptide corresponding to 17 amino acids near the carboxy terminus of human ADAM10. The immunogen is located within the last 50 amino acids of ADAM10.
• Concentration: 1 mg/mL
• Dilution range: WB: 1 μg/mL; IHC: 2 μg/mL; IF: 10 μg/mL.Antibody validated: Western Blot in human, mouse and rat samples; Immunofluorescence in human, mouse and rat samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 84kDObserved: 94 kD (precursor) and 68kD (mature form)
• Target: ADAM10
• UniProt ID: O14672
• NCBI: NP_001101
• Storage: ADAM10 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: ADAM10 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: ADAM10 Antibody: RAK, kuz, AD10, AD18, MADM, CD156
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

ADAM10 KO Validation in MEF Cells. Loading: 10 µg of lysate Antibodies: ADAM10 orb1239162, 1 µg/mL and beta-actin, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. orb1239162 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

ADAM10 KO Validation in 293 Cells. Loading: 15 µg of lysate Antibodies: ADAM10 orb1239162, 2 µg/mL and beta-actin, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. orb1239162 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 orb1239162, 0.5 µg/mL, ADAM10 orb1255477, 1 µg/mL, and -actin orb1240312, 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 orb1239162, 1 µg/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution. orb1239162 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Western Blot Validation in Mouse Tissues. Loading: 15 µg of lysates per lane. Antibodies: ADAM10 orb1239162, 1 µg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. orb1239162 detected both precursor ADAM10 (94KD) and mature ADAM10 (68kD).

Western Blot Validation in Rat Tissues. Loading: 15 µg of lysates per lane. Antibodies: ADAM10 orb1239162, 1 µg/mL, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. orb1239162 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 orb1239162 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 K562 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling ADAM10 with orb1239162 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 orb1239162 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 orb1239162 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

WB Validation of ADAM10 in Human washed platelets (Al-Tamimi et al., 2012). Human washed platelets (5x 108/mL) were lysed and ADAM10 expression was detected byanti-ADAM10 cytoplasmic tailantibody (ADAM10 CT). Only mature ADAM10 (~72kD), when got cleaved upon activation, was detectable in resting platelet lysates. Vertical lines indicate a repositioned lane.

Regulated Expression Validation of ADAM10 in Rat cortical cultures (Hurtado et al., 2002). Western Blot analysis was carried out to monitor protein expression of ADAM10 with anti-ADAM10 antibodies. ADAM10 expression was decreased in rat cortical cultures when exposed to oxygen-glucose deprivation (OGD) or glutamate.

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 thehuman 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.