EPRS1/PARS Antibody (monoclonal, 12B5B3)

• Catalog Number: orb865669
• Category: Antibodies
• Description: EPRS1/PARS Antibody (monoclonal, 12B5B3)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 12B5B3
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human
• Isotype: Mouse IgG2b
• Immunogen: E.coli-derived human EPRS1/PARS recombinant protein (Position: R1298-Y1512).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 170-180 kDa
• UniProt ID: P07814
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of EPRS1/PARS using anti-EPRS1/PARS antibody (orb865669). 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 30 ug of sample under reducing conditions. Lane 1: human 293T whole cell lysates, Lane 2: human K562 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-EPRS1/PARS antigen affinity purified monoclonal antibody (Catalog # orb865669) 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 EPRS1/PARS at approximately 170-180 kDa. The expected band size for EPRS1/PARS is at 171 kDa.

IHC analysis of EPRS1/PARS using anti-EPRS1/PARS antibody (orb865669). EPRS1/PARS was detected in a paraffin-embedded section of human testis cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml mouse anti-EPRS1/PARS Antibody (orb865669) 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.

IF analysis of EPRS1/PARS using anti-EPRS1/PARS antibody (orb865669). EPRS1/PARS was detected in an immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-EPRS1/PARS Antibody (orb865669) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of CACO-2 cells using anti-EPRS1/PARS antibody (orb865669). Overlay histogram showing CACO-2 cells stained with orb865669 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-EPRS1/PARS Antibody (orb865669, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.