Chemiluminescent Western Blot Kit

• Catalog Number: orb342274
• Category: Assays and Kits
• Description: Chemiluminescent Western Blot Kit
• Tested applications: WB
• Dilution range: WB: User Optimized
• Purity: This Chemiluminescent Western Blot Kit allows for the detection of FLAG -tagged recombinant protein present in cell lysates provided by the user.
• Conjugation: Unconjugated
• Hazard Information: Non-Toxic
• Storage: See kit insert for complete instructions.
• Buffer/Preservatives: Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity!
• Alternative names: Flag Western Blot, Western Blotting Kit, Chemilumi
• Note: For research use only
• Application notes: Use Rockland Immuno-chemicals’ Anti-DYKDDDDK (FLAG) Chemiluminescent Kit for Western Blotting for detection of FLAG -tagged recombinant proteins by Western blot. This kit is useful for both ''Western blotting'' and ''dot blotting'' methods. After protein separation and transfer, the membrane is probed with Anti-DYKDDDDK (FLAG). Detection of the membrane bound antibody-antigen complex is achieved by the addition of a secondary antibody conjugated to the enzyme horseradish peroxidase. The enzyme reacts with a specialized formulation of luminol, an extremely sensitive, non-radioactive substrate that emits light and allows visualization using X-ray film or other imaging methods, including highly sensitive CCD cameras and imaging systems.
• Expiration Date: Please enquire.

Western Blot. Anti-DYKDDDDK (FLAG®) Chemiluminescent Kit for Western blot was used to detect known amounts of bacterial alkaline phosphatase (BAP) protein containing the FLAG® epitope tag. N-terminal linked and C-terminal linked BAP were diluted into a HeLa whole cell lysates. After separation by SDS-PAGE lysates were transferred to a membrane which was blocked with 5% BLOTTO (p/n orb348624) in TTBS for 1 h at 4 °C. The blot was probed with Anti-DYKDDDDK (FLAG®) followed by HRP anti-Mouse IgG (p/n orb347385) at 1:10000 dilution in blocking buffer for 1 h at 4°C.