Myosin light chain kinase MYLK Rabbit Monoclonal Antibody

• Catalog Number: orb547832
• Category: Antibodies
• Description: Myosin light chain kinase MYLK Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: G-13
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human Myosin light chain kinase
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 210715 MW
• UniProt ID: Q15746
• 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.
• Alternative names: Myosin light chain kinase, smooth muscle;MLCK;smML
• Note: For research use only
• Application notes: WB 1:500-1:2000IHC 1:50-1:200ICC/IF 1:50-1:200FC 1:50
• Expiration Date: 12 months from date of receipt.

Immunohistochemical analysis of paraffin-embedded human colon, using Myosin light chain kinase Antibody (orb547832) MYLK was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-MYLK Antibody (orb547832) overnight at 4 Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

Western blot analysis of myosin light chain kinase expression in HUVEC cell lysate (orb547832). 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. 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 rabbit anti-MYLK monoclonal antibody (Catalog # orb547832) overnight at 4 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 1:10000 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 MYLK.