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A8181

Sigma-Aldrich

Amido Black Staining Solution 2X

electrophoresis reagent

Synonym(s):

Naphthol Blue Black solution, Amido Black 10B

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About This Item

Empirical Formula (Hill Notation):
C22H14N6Na2O9S2
CAS Number:
1064-48-8
Molecular Weight:
616.49
Beilstein:
5374263
MDL number:
MFCD00004017
UNSPSC Code:
12171500
PubChem Substance ID:
24891210
NACRES:
NA.32
Pricing and availability is not currently available.

Quality Level

200

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

Amido black (AB) is an acidic dye used for staining proteins.[1] It can detect proteins levels of 50ng in both polyvinylidene difluoride (PVDF) and nitrocellulose membrane.[2] AB aids protein visualization even at low concentrations.[3] It has applications in forensics due to its ability to stain blood proteins in fingerprints.[1]

Application

Amido Black Staining Solution 2X has been used in the staining:
  • serum proteins present in the culture supernatant[4]
  • cysteine sulfinic acid decarboxylase protein in mouse liver homogenate[5]
  • histones from chromatin samples[6]

Reconstitution

After dilution with deionized water to a final volume of 500 ml, solution will contain 0.1% (w/v) amido black, 25% (v/v) isopropanol and 10% (v/v) acetic acid.

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Product No.
Description
Pricing

Pictograms

FlameExclamation mark
GHS02,GHS07

Signal Word

Danger

Hazard Statements

H225,H315,H319,H336

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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Certificates of Analysis (COA)

Lot/Batch Number
SLCP7209
SLCK9090
SLCF5171
SLBW0939
SLBR9882V

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Placental release of taurine to both the maternal and fetal circulations in human term pregnancies
Holm MB, et al.
Amino Acids, 50(9), 1205-1214 (2018)
Detection of proteins on blot membranes
Goldman A, et al.
Current Protocols in Protein Science / Editorial Board, John E. Coligan ... [Et Al.], 86(1), 10-18 (2016)
Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to a positively charged membrane filter.
J M Gershoni et al.
Analytical biochemistry, 124(2), 396-405 (1982-08-01)
Meghan C Hughes et al.
The Journal of physiology, 598(7), 1377-1392 (2019-01-24)
Ninety-eight per cent of patients with Duchenne muscular dystrophy (DMD) develop cardiomyopathy, with 40% developing heart failure. While increased propensity for mitochondrial induction of cell death has been observed in left ventricle, it remains unknown whether this is linked to
Protein Blotting and detection: methods and protocols, 86(1), 10-18 (2009)
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Protein Staining Reagents Selection Guide

Sigma offers EZBlue™ and ProteoSilver™ reagents for protein visualization, suitable for proteomics and traditional PAGE formats.

Protein Staining Reagents Selection Guide

Sigma offers EZBlue™ and ProteoSilver™ reagents for protein visualization, suitable for proteomics and traditional PAGE formats.

Protein Staining Reagents Selection Guide

Sigma offers EZBlue™ and ProteoSilver™ reagents for protein visualization, suitable for proteomics and traditional PAGE formats.

Questions

  1. 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?

    1 answer

    1. 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.

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