b'SuperSignal West Atto Ultimate Sensitivity Substrate Features:Thermo Scientific SuperSignal West Atto Ultimate Sensitivitylow-femtogram to high-attogramSensitivity Substrate is an ultrasensitive ECL substrate that enables protein detection at high-attogram levels by Stability48 hours for working solution, one year for kit western blot analysis with the HRP enzyme. It providesat 4Chigher-level sensitivity and better signal-to-noise ratios Compatibilitynitrocellulose and PVDF membranesthan other commercially available high-performance HRP substrates (Figure 30). It is the ideal choice for detection Signal durationup to 6 hoursof very low-abundance targets or when using precious Recommended primary antibody samples that require maximum levels of sensitivity. concentration1:1,0001:5,000 dilution(0.21.0 g/mL) Recommended secondary antibody concentration1:100,0001:250,000 dilution (410 ng/mL)5 2.5 1.25 0.625 0.1560.312A BgSuperSignal West AttoSuperSignal West Atto 2.2 Bio-Rad Clarity Max2.0 Amersham ECL SelectBio-Rad Clarity Max Immobilon WesternSignal-to-noise ratio1.8 Bio-Rad ClarityAmersham ECL Select Standard ECL1.6Immobilon Western1.4Bio-Rad Clarity1.2Standard ECL1.00.625 g 0.313 g 0.156 gHeLa lysate proteinFigure 30. Chemiluminescent substrate product comparison with SuperSignal West Atto substrate. (A) Akt (pan) detection in HeLa lysate was performed using SuperSignal West Atto, Bio-Rad Clarity Max, Amersham ECL Select, Millipore Immobilon Western, Bio-Rad Clarity, and standard ECL substrates. The blots were developed using Akt (pan) monoclonal antibody, followed by HRP-conjugated goat antirabbit IgG secondary antibody. Blots were incubated with respective substrates per product instructions. Images were captured using the iBright Imaging System. (B) The signal-to-noise ratios for the western blot bands in the boxed area for SuperSignal West Atto, Bio-Rad Clarity Max, Amersham ECL Select, Millipore Immobilon Western, Bio-Rad Clarity, and standard ECL substrates are shown in bar graph form. SuperSignal West Atto substrate provided the best signal-to-noise ratios across all protein loads. The standard ECL substrate didnt have detectable signal above background noise after 0.625 g. Clarity and Immobilon substrates didnt have detectable signal above background noise after 0.313 g.50'