TrueBlack® WB Blocking Buffer Kit | 고마바이오텍

특징

• 형광을 이용한 western blot시, 비특이적 결합과 백그라운드 형광을 최소화합니다.
• LI-COR사의 Odyssey® Blocking Buffer에 비해 성능이 좋고 경제적입니다. 
• BSA, gelatin, casein에 비해 charged dye로 인한 백그라운드를 잘 억제합니다.
  
• PVDF 및 NC (nitrocellulose) Membrane에 사용할 수 있습니다.
• 여러 species의 항체에 적용할 수 있습니다. 
• Phosphoprotein detection에도 적용할 수 있습니다.
  
• 포유류 단백질을 포함하지 않으므로, anti-goat, anti-bovine 등 다양한 2차 항체와 함께 사용할 수 있습니다.
• 가시광선과 근적외선 형광 western blot에 사용할 수 있습니다.
• 참고: HRP-based chemiluminescent detection에는 사용할 수 없습니다.

구성 

• TrueBlack® WB Blocking Buffer
• TrueBlack® WB Antibody Diluent

Figure 1. Western detection of tubulin in HeLa cell lysate with mouse anti-tubulin and Alexa Fluor® 790 goat anti-mouse antibodies. Membranes were blocked with fish gelatin blocking buffer or TrueBlack® WB Blocking Buffer. The highly negatively charged Alexa Fluor® 790 labeled antibody showed non-specific binding to the PVDF membrane and cellular proteins, which was blocked by TrueBlack® WB Blocking Buffer. Lanes 1-3: 10 ug, 1 ug, or 0.1 ug HeLa cell total protein. 

Note: Alexa Fluor ®790 dye was used to demonstrate non-specific background from highly charged dyes. Biotium's near-IR CF® dyes are peggylated, a structural feature that renders the dyes highly water soluble without excessive negative charge, resulting in significantly lower background.

 

Figure 2. Western detection of phospho-Erk1/2 in PDGF-stimulated NIH-3T3 cell lysate. Membranes were blocked with fish gelatin blocking buffer (Gelatin), LI-COR® Odyssey® TBS Blocking Buffer (Odyssey®), or TrueBlack® WB Blocking Buffer (TrueBlack® WB). Phosphorylated extracellular kinases (pERK, showing two isoforms) were detected using rabbit anti-pERK primary antibody and CF® 680R-Donkey anti-rabbit secondary antibody. The membrane blocked with TrueBlack® WB Blocking Buffer shows lower background fluorescence and better specificity compared to the other membranes.