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Blotting Reagents Kits on ZAGENO

Blotting reagents are essential in western, northern and southern blotting for sensitive and consistent detection of proteins and nucleic acids from complex samples. These reagents improve the efficiency of electrophoresis and, with the help of antibodies, enable radiometric, chemiluminescent, chromogenic, and fluorescent detection for use in the fields of molecular biology and immunogenetics.

Expand your understanding in our Knowledge Section. Perfect your experiments with our Troubleshooting Guides. Join in the discussion within our Community. Learn about ZAGENO at How It Works.

Western Blotting

Western blotting separates proteins on a gel by electrophoresis before transferring them to a membrane, upon which the proteins are detected by chemiluminescence, fluorescence or colorimetry.

See how Western Blotting is being used in breakthrough research in our article: Are Antibody Vaccines the answer to Alzheimer's Disease?

Southern Blotting

Southern Blotting uses electrophoresis to separate DNA fragments which are transferred to a positive nylon membrane.

Northern Blotting

Northern Blotting separates RNA fragments via electrophoresis before transferring the resultant bands to a positively charged nylon membrane.

Blotting Reagents Products

Blotting Reagents for these processes include:

Transfer Buffers
Transfer buffers help shift the sample over from the gel used in electrophoresis to the detection membrane.

Wash and Blocking Buffers
Wash buffers are repeatedly used during the blotting process. For example, in western blotting, between antibody incubation steps, wash buffers are used to remove any excess antibodies between primary and secondary antibody incubation steps. During northern and southern blotting, washing buffers remove unhybridized probes.

Blocking buffers, on the other hand, prevent the antibodies from nonspecific binding.

Filter / Blotting Paper
The filter paper is necessary for use in the transfer sandwich to enable quick and efficient transfer by supporting wicking and uniform capillary action, as well as preventing contamination.

Blotting Membranes
The blotting membrane (used for detection) consists of either nitrocellulose or PVDF (polyvinylidene fluoride). Both membranes are suitable for use in Western Blots, while PVDF is also capable of binding nucleic acids well, so it can be used in northern, southern and dot blots as well. However, northern and southern blotting also use positively charged nylon membranes, which can permanently bind nucleic acids with low background and high target retention.

Detection Reagents
Detection reagents include chemiluminescent substrates, chromogenic substrates and fluorescence detection kits. All these reagents interact with a probe on the antibody bound to the protein of interest (western) or a hybridization probe (northern/southern) to provide a visual signal.

Chemiluminescent substrates have the greatest sensitivity, able to quantify concentration and can be used after reprobing. They require the use of an x-ray film and dark room to detect the light emission created by a reaction between two solutions.

Chromogenic substrates do not need any visualization equipment because they form an insoluble colored precipitate on contact with the appropriate enzyme. However, this method lacks sensitivity.

Fluorescent kits can quantitatively analyze a blot without a darkroom or film and can be used to target multiple, different samples on the same blot without reprobing.

Compare Western Transfer Buffers

With our compare function, you can avoid all the time and energy wasted sifting through multiple web pages from different suppliers. At ZAGENO you can clearly see kits side-by-side, with the relevant attributes for each kit neatly in line for easy selection of the best product for you.

Click on the comparison below for a clearer view!

For example, this comparison shows:

  • that the G10X TRIS/CAPS BUFFER Kit from Bio-Rad is specifically formulated for semi-dry blotting.
  • that the EFFICIENT™ WESTERN TRANSFER BUFFER [20X] Kit from G-Biosciences is twice as concentrated.
  • that the 10X TRIS/GLYCINE BUFFER Kit from Bio-Rad is the cheapest per ml.

This comparison clearly exemplifies how every kit has strengths and weaknesses. Depending on what features you require for your experiment, these details should enable you to make an informed decision on the right kit for you.

The ZAGENO comparison does not highlight one kit to be better than the other, as the kit of choice may vary between researchers - depending on each individual's preferred attributes. The best kit is the one that meets your needs - ZAGENO allows you to make an informed decision with minimum effort.

Check out our How It Works page for a guide to using the comparison function.

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