To detect total protein on a Western blot, follow these steps:
1.Prepare Amido Black 1X Staining Solution by diluting Amido Black Staining Solution (Product Number A8181) 1:1 with deionized water.
2. After transferring proteins onto nitrocellulose membrane, immerse membrane in sufficient Amido Black 1X Staining Solution to cover. Stain for 1 minute.
3. Destain the membrane by placing in an aqueous solution containing 25% isopropanol and 10% acetic acid for 30 minutes. If a lighter background is desired, destain overnight.
4. Store the destained membrane in deionized water or allow to air dry.
Product Number A8181 Store at Room Temperature Product Description: Amido Black staining solution is used for the detection of microgram quantities of proteins transferred to nitrocellulose membranes. In comparison to staining with Brilliant Blue R, staining with Amido Black results in significantly less background on nitrocellulose membranes. Because nitrocellulose membranes are more stable in isopropanol than in methanol or other solvents, the formulation contains isopropanol. After a 1:1 dilution with deionized water, Amido Black Staining Solution contains: 0.1% Amido Black (w/v), 25% Isopropanol (v/v), 10% Acetic Acid (v/v). Note: Not suitable for use with charge modified nylon membranes.
A8181
Amido Black Staining Solution 2X
electrophoresis reagent
Synonym(s):
Naphthol Blue Black solution, Amido Black 10B
About This Item
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Quality Level
form
liquid
SMILES string
[Na+].[Na+].Nc1c(\N=N/c2ccc(cc2)[N+]([O-])=O)c(cc3cc(c(\C=C/c4ccccc4)c(O)c13)S([O-])(=O)=O)S([O-])(=O)=O
InChI
1S/C22H16N6O9S2.2Na/c23-19-18-12(11-17(39(35,36)37)21(22(18)29)27-24-13-4-2-1-3-5-13)10-16(38(32,33)34)20(19)26-25-14-6-8-15(9-7-14)28(30)31;;/h1-11,29H,23H2,(H,32,33,34)(H,35,36,37);;/q;2*+1/p-2/b26-25+,27-24+;;
InChI key
AOMZHDJXSYHPKS-DROYEMJCSA-L
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General description
Application
Reconstitution
related product
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Flam. Liq. 2 - Skin Irrit. 2 - STOT SE 3
Target Organs
Central nervous system
Storage Class Code
3 - Flammable liquids
WGK
WGK 1
Flash Point(F)
53.6 °F
Flash Point(C)
12 °C
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Could you please provide the staining procedure and destaining formulation for the Amino Black Staining solution 2x that is suitable for staining in 2-40% gels?
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