heavy chain Myosin/Myh3 Antibody

• Catalog Number: orb865435
• Category: Antibodies
• Description: heavy chain Myosin/Myh3 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived rat heavy chain Myosin/Myh3 recombinant protein (Position: K44-S119).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5 μg/ml, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat Flow Cytometry, 1-3 μg/1x106 cells,Human, Rat Direct ELISA, 0.1-0.5 μg/ml, Rat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 224 kDa
• UniProt ID: P12847
• Storage: 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 freezing and thawing.
• 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 heavy chain Myosin/Myh3 using anti-heavy chain Myosin/Myh3 antibody (orb865435). 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: rat heart tissue 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 rabbit anti-heavy chain Myosin/Myh3 antigen affinity purified polyclonal antibody (Catalog # orb865435) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for heavy chain Myosin/Myh3 at approximately 224 kDa. The expected band size for heavy chain Myosin/Myh3 is at 224 kDa.

IHC analysis of heavy chain Myosin/Myh3 using anti-heavy chain Myosin/Myh3 antibody (orb865435). heavy chain Myosin/Myh3 was detected in a paraffin-embedded section of mouse cardiac 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 rabbit anti-heavy chain Myosin/Myh3 Antibody (orb865435) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.

IHC analysis of heavy chain Myosin/Myh3 using anti-heavy chain Myosin/Myh3 antibody (orb865435). heavy chain Myosin/Myh3 was detected in a paraffin-embedded section of rat cardiac 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 rabbit anti-heavy chain Myosin/Myh3 Antibody (orb865435) overnight at 4°C. Biotinylated goat anti-rabbit 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 # orb90444) with DAB as the chromogen.

Flow Cytometry analysis of U20S cells using anti-heavy chain Myosin/Myh3 antibody (orb865435). Overlay histogram showing U20S cells stained with orb865435 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-heavy chain Myosin/Myh3 Antibody (orb865435, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.