Western transfer, or Western Blotting, involves protein transfer from polyacrylamide gels to a PVDF or nitrocellulose membrane to allow detection by specific antibodies. G-Biosciences offers high quality reagents and resources for the entire Western transfer process to ensure successful transfer and clean protein detection.
For the initial transfer, G-Biosciences has high protein binding membranes (PVDF and nitrocellulose) and high efficiency Western Transfer Buffers, including a buffer specifically designed for high molecular weight protein transfer. Following protein transfer, it’s highly recommend to check protein transfer efficiency with a rapid and reversible protein stain, such as our Swift™ Membrane Stain or BLOT-FastStain™. For improved protein transfer or for difficult to transfer proteins, such as high molecular weight proteins, G-Biosciences has Western Transfer Pads for improved transfer efficiency.
For probing the Western Transfer Membranes, the first step is to block non-specific binding sites with an appropriate blocking agent. G-Biosciences offers a whole panel of blocking agents, including non animal protein blocking agents (NAP-BLOCKER™ or Protein-Free™ blocking agent), non-sera animal protein blocking agents (Superior™ Blocking Agent or FISH-Blocker™, and standard blocking solutions using milk, casein or BSA. A convenient Blocking Agents Trial Pack is offered to determine the best blocking agent to use.
Once the membrane is blocked, a primary antibody of choice is added. G-Biosciences offers over 21,000 primary antibodies that can all be directly conjugated to HRP or other detection probes. Alternatively, a secondary antibody conjugated to horseradish peroxidase (HRP) or alkaline phosphatase (AP) can be used to detect the primary antibody. Following incubation of the primary and secondary antibodies, the membranes need to be extensively washed with wash buffers (TBS-Tween or PBS-Tween).
The final step is protein detection, where the detection probe is visualized via chemiluminescence or chromogenic reactions. Our femtoLUCENT™ PLUS and picoLUCENT™ PLUS are highly sensitive chemiluminescence reagents for HRP and AP, whereas femtoCHROMO™-AP and femtoCHROMO™-HRP allow for chromogenic detection of proteins.
Our Western ReProbe™ products allow you to strip your blot and reprobe with different antibodies, such as housekeeping antibodies, and our and Swift™ Film Cleaner helps clean spotty and overexposed films.
Visit our partner, BT Lab Systems, for Protein Electrophoresis and Western Blotting equipment, including semi dry transfer units.