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Revision as of 08:53, 10 September 2015

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Amido black assay

Protein concentration Amido black assay

material: Amido black solution: 10% (v/v)Acectic acid, 90% (v/v)MeOH, 0.05% (v/v) Amido black; Destain: 90% (v/v)EtoH; 10% (v/v) Acetic acid …
duration: 1h

  • transfer 20ul of protein extract into new Eppendorf tubes; pipet samples for the calibration curve (see table 1)
  • add 1 ml amido black solution; vortex
  • centrifuge for 15 min at max. speed, RT
  • discard supernatant
  • add 1 ml destain solution
  • mix by inversion
  • centrifuge for 15 min at max. speed, RT
  • discard supernatant
  • air-dry the pellet (takes about 15 min)
  • add 1ml 0.2 M NaOH; vortex
  • transfer the samplkes to disposable cuvettes
  • measure the OD620
  • calculate the concentration of your protein samples
1 µg/µl BSA µg BSA
5µl 5µg
10µl 10µg
15µl 15µg
20µl 20µg
30µl 30µg
,