ADK Antibody (monoclonal, 7F4)

• Catalog Number: orb547973
• Category: Antibodies
• Description: ADK Antibody (monoclonal, 7F4)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 7F4
• Tested applications: FC, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: E. coli-derived human ADK recombinant protein (Position: K165-T351). Human ADK shares 88.8% and 88.2% amino acid (aa) sequence identity with mouse and rat ADK, respectively.
• Concentration: 0
• Dilution range: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Flow Cytometry, 1-3μg/1x106 cells
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 45 kDa
• UniProt ID: P55263
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: Adenosine kinase; AK; Adenosine 5'-phosphotransfer
• Note: For research use only
• Application notes: Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of ADK using anti-ADK antibody (orb547973). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human PC-3 whole cell lysates, Lane 2: human U-87MG whole cell lysates, Lane 3: human SW620 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-ADK antigen affinity purified monoclonal antibody (Catalog # orb547973) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for ADK at approximately 45KD. The expected band size for ADK is at 45KD.

IHC analysis of ADK using anti-ADK antibody (orb547973). ADK was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-ADK Antibody (orb547973) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of ADK using anti-ADK antibody (orb547973). ADK was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-ADK Antibody (orb547973) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

IHC analysis of ADK using anti-ADK antibody (orb547973). ADK was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-ADK Antibody (orb547973) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90443) with DAB as the chromogen.

Flow Cytometry analysis of PC-3 cells using anti-ADK antibody (orb547973). Overlay histogram showing PC-3 cells stained with orb547973 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ADK Antibody (orb547973, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.