Thermo Scientific SuperSignal West Femto Maximum Sensitivity Substrateは、西洋ワサビペルオキシダーゼ(HRP)酵素によるウェスタンブロッティングで低フェムトグラムレベルのタンパク質を検出するための、超高感度の高度化学発光(ECL)基質です。
SuperSignal West Femto Maximum Sensitivity Substrateの特性: •高感度: 低フェムトグラム • 安定性:溶液の効果は8時間持続し、キットは4℃で1年間安定 • 適合性:ニトロセルロース膜およびPVDF膜に対応 • シグナル持続時間:最長8時間 • 一次抗体の推奨濃度:希釈比1:5,000~1:100,000(10~200 ng/mL) • 二次抗体の推奨濃度:希釈比1:100,000~1:500,000(2~10 ng/mL)
SuperSignal West Femto Maximum Sensitivity Substrateは、ウェスタンブロット解析による低フェムトグラムレベルの検出を実現するよう設計されています。SuperSignal West Femto基質は、最適な抗体濃度およびブロッキングバッファーと組み合わせることで、標準的なECL基質では確認できない微量の標的タンパク質の検出を可能にします。
対応範囲:12ミニゲルブロット、1000 cm2相当のメンブレン • ルミノール/エンハンサー、50 mL • 安定した過酸化物バッファー、50 mL
試薬は4℃で保存。
よくあるご質問(FAQ)
What types of membrane work best with SuperSignal West Femto Maximum Sensitivity Substrate?
Most researchers use nitrocellulose or polyvinyldiflouride (PVDF) membranes with SuperSignal West Femto Maximum Sensitivity Substrate. Both work well, although nitrocellulose seems to be better suited in some applications than PVDF. In addition, charge-modified nylon membrane performs well with this substrate. Please also see our guide for choosing western blot membranes (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/western-blotting/transfer-proteins-western-blot/membranes-transfer-buffers-western-blotting/membranes-western-blotting.html).
Will SuperSignal West Femto Maximum Sensitivity Substrate work with nucleic acid blots (Southern blotting, etc.)?
Yes. However, SuperSignal West Femto Maximum Sensitivity Substrate was optimized for use in Western blot sand is generally not sensitive enough for most nucleic acid protocols. For Southern and Northern blotting, use our North2South Chemiluminescent Nucleic Acid Hybridization and Detection Kit (Cat. No. 17097), North2South Chemiluminescent Substrate for HRP (Cat. No. 17295), or the Chemiluminescent Nucleic Acid Detection Kit, if probing for biotinylated probes (Cat. No. 89880).
I used the SuperSignal West Femto Maximum Sensitivity Substrate. Why is the entire film black or showing a reversed image?
The antibody concentration (primary and secondary) is too high. Use the antibody dilutions described in the product instructions. Most background will disappear when the proper blocking reagent and a HRP conjugate concentration are used.
Why do I see more bands with the SuperSignal West Femto Maximum Sensitivity Substrate than I've seen in the past with other substrates?
Because this substrate is more sensitive than other chemiluminescent substrates you might detect low-abundance proteins that were not visible before. When using a more sensitive substrate, more careful optimization of blocking buffer steps and antibody concentrations is required.