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Western Transfer

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.

Protein Transfer

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.

Western Blotting

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).

Protein Detection

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.


Download the Western Blotting Handbook for Troubleshooting Tips
femtoLUCENT™ PLUS is a femtogram level sensitive chemiluminescence immunodetection system that allows users the option to detect even hard-to-detect low abundance proteins. This immunodetection system is based on an innovative formulation of luminol and 1, 2 dioxetane, a supersensitive detecti..
A 10X Concentrated solution of Phosphate Buffered Saline with Tween® 20. The 10X Concentration is 80mM Na2HPO4, 1.5M NaCl, 20mM KH2PO4, 30mM KCl, 0.5% Tween® 20, pH 7.4. A dry format PBST is also avaialble PBST is commonly used as a wash solution for Western blot membranes and microt..
Swift™ Membrane Stain is a unique, proprietary (patents pending), rapid, reversible, ready-to-use membrane stain for proteins on nitrocellulose or PVDF membranes. Swift™ Membrane Stain stains proteins faster and with 500 times more sensitivity than the routinely used Ponceau-S stain and ..
Swift Film Cleaner™ allows researchers to clean film that has been overexposed or has a high background or speckling without having to repeat experiments.  The Swift Film Cleaner™ is suitable for all experiments that use film to develop the result, including gel shift assays, Wester..
$0.00 provides an interactive version of this protocol where you can discover and share optimizations with the research community.  Western blot analysis of proteins is a routine and commonly used technique in research laboratories, with 3 major drawbacks. The first is the amount of tim..
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