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o-Phenylenediamine dihydrochloride SKU: P6662
Reagent Type
Other reagents
Other reagents
Blot Type
/
Detection Technique
/
Scientific Score
4.64
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 141.00 - $ 284.10
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o-Phenylenediamine dihydrochloride SKU: P6787
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
6.28
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 141.00 - $ 264.12
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o-Phenylenediamine dihydrochloride SKU: P6912
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.78
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 163.00 - $ 301.85
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o-Phenylenediamine dihydrochloride SKU: P7288
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
6.37
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 201.00 - $ 372.88
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Protein G−Peroxidase from Streptococcus sp. SKU: P8170
Reagent Type
Other reagents
Other reagents
Blot Type
/
Detection Technique
/
Scientific Score
5.73
Brand
Sigma-Aldrich
Sizes
1 (250 µg)
Available From
2 Suppliers
Price Range
$ 123.00 - $ 126.90
Catalog IDs
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o-Phenylenediamine dihydrochloride SKU: P8287
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.60
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 187.00 - $ 347.35
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o-Phenylenediamine dihydrochloride SKU: P8412
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.72
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 210.00 - $ 387.30
Added to comparison remove item
o-Phenylenediamine dihydrochloride SKU: P8787
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.74
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 175.00 - $ 325.16
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o-Phenylenediamine dihydrochloride SKU: P8806
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.60
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 Tablets)
Available From
2 Suppliers
Price Range
$ 157.00 - $ 296.30
Added to comparison remove item
o-Phenylenediamine SKU: P9029
Reagent Type
Substrates
Substrates
Blot Type
/
Detection Technique
/
Scientific Score
7.78
Brand
Sigma-Aldrich
Sizes
2 (50 - 100 g)
Available From
2 Suppliers
Price Range
$ 172.00 - $ 340.70
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SignalStain® Antibody Diluent
Reagent Type
Antibody diluent
Antibody diluent
Blot Type
/
Detection Technique
/
Scientific Score
9.57
Brand
Cell Signaling Technology
Sizes
2 (25000 - 100000 µl)
Available From
1 Supplier
Price Range
$ 40.00 - $ 82.00
Catalog IDs
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Nytran N Nylon Blotting Membrane, 0.2 μm, 20 cm x 3 m
Reagent Type
Blotting membranes
Blotting membranes
Blot Type
/
Detection Technique
/
Scientific Score
5.75
Brand
Whatman
Sizes
1 (1 Roll)
Available From
2 Suppliers
Price Range
$ 426.00 - $ 494.14
Catalog IDs
Added to comparison remove item

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Applications in Blotting

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.

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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.