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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
A unique stain for reversibly staining protein on nitrocellulose and PVDF transfer membranes.   BLOT-FastStain™ only stains protein and leaves the background absolutely untouched and brilliant white resulting in high band visibility. The staining procedure takes 10 minutes and has a sen..
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G-Biosciences scientists have developed a high efficiency transfer buffer for Western blotting. The pre-made and ready-to-use [20X] transfer buffer is prepared for optimal conductivity and efficient protein transfer without generating excessive heat or transfer distortion. Efficient™ Western T..
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femto TBST™ and femto PBST™ will enhance sensitivity of your immunoassays by minimizing the "washing out" of immuno agents or immuno complexes, a common problem associated with classical TBST and PBST buffers. The wash buffers aid in the generation of cleaner backgrounds, resul..
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femtoCHROMO™ AP kit is supplied with a single step chromogenic substrate for the rapid development of protein bands with low background staining. To ensure low background staining, each kit is supplied with an enhanced blocking agent, BLOT-QuickBlocker™, and a concentrated (10X) was..
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femtoCHROMO™-HRP kit is supplied with a single step chromogenic substrate for the rapid development of protein bands with low background staining.  To ensure low background staining, each kit is supplied with an enhanced blocking agent, BLOT-QuickBlocker™, and a concentrated (10X) w..
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The femtoLUCENT™ PLUS-AP kit is based on alkaline phosphatase (AP) chemiluminescence reaction. The chemiluminescence light emission can be recorded by a short exposure to blue-light sensitive autoradiography films. The femtoLUCENT™ PLUS-AP kit allows for detection of femtogram quantity o..
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FirstChoice™ Blocking Buffers contain a proprietary protein formulation that offers greater versatility and lack of cross-reactivity.  This makes FirstChoice™ Blocking Buffers ideal as a first choice for optimization of new assays, system or when determining the optimal blocking buf..
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FISH-Blocker is a blocking agent that uses a fish protein as the primary blocking agent.  The use of a fish protein, a non-mammalian protein, is that it eliminates or minimizes the interaction of antibodies raised in mammals.  FISH-Blocker is one of the best blocking agents for immunoassay..
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For the highly sensitive detection of glycoproteins following gel electrophoresis (Figure 1) or protein transfer to nitrocellulose membranes (Figure 2).   The kit uses an enhanced Periodic Acid-Schiff (PAS) method for detection of glycoprotein sugars.  The supplied oxidizing agent fi..
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Based on Efficient™ Western Transfer Buffer, the High Molecular Weight Transfer buffer is designed to facilitate the transfer of notoriously difficult high molecular weight proteins (>70kDa) during Western blotting.   Supplied as 1 liter of a 5X concentrated solution...
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For user's convenience, we offer pre-cut transfer membranes and padding for Western blot transfer procedures. Pre-cut membranes are supplied sandwiched between blotting paper padding. Simply soak the membrane in transfer buffer and assemble with the gel in a transfer cassette. Nitrocellulose, (0..
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picoLUCENT™ PLUS-AP kit is based on our ultra sensitive Luminol substrate that produces chemiluminescence upon reaction with horseradish peroxidase. The chemiluminescence light emission can be recorded by a short exposure to autoradiography films.    picoLUCENT™ PLUS-AP is al..
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