Restore PLUS BufferはオリジナルのThermo Scientific Restore Western Blot Stripping Bufferの代替製剤です。Restore PLUS Stripping Bufferは、抗体をウェスタンブロットから除去するのが困難で、より長時間のインキュベーションを要するか、またはより穏やかな製剤では22℃以上でインキュベーションする必要ある場合に使用するよう設計されています。高親和性の抗体を室温で迅速かつ効果的にウェスタンブロットから除去でき、転写タンパク質は除去されないためターゲットを何度も再プローブ可能です。
プロトコルの概要: • ブロットを洗浄し、化学発光基質を除去する • ブロットをRestore Western Blot Stripping Bufferで5~15分間室温でインキュベートする (高親和性の抗体の場合は、37°でインキュベートする) • ブロットを取り出し、Wash Bufferで洗浄する • 膜をブロックする • 抗体が十分除去されていることを確認する • 次の免疫ブロット実験を実行する
研究用途にのみご使用ください。診断目的には使用できません。
仕様
検出法Chemiluminescence
使用対象(アプリケーション)Western Blot
形状液体
Membrane CompatibilityNitrocellulose, PVDF
製品ラインRestore
製品タイプWestern Blot Stripping Buffer
数量500 mL
Unit SizeEach
組成および保存条件
受け取り後、室温で保存。製品は室温で出荷されます。
よくあるご質問(FAQ)
How many times can I strip my blot using Restore or Restore Plus Western Blot Stripping Buffers ?
The stability of the attached (transferred and bound) protein will determine the number of times the membrane can be successfully re-probed after stripping. The protein may withstand stripping as many as four times or as few as one time.
Can I strip membranes with Restore or Restore Plus Western Blot Stripping Buffer and reprobe for subsequent detection with fluorescent probes?
No. Although the fluorescent antibodies, like other antibodies, can be stripped with Restore Buffers, stripped membranes typically produce unacceptable background for subsequent fluorescent detection methods. We recommend Restore Fluorescent Western Blot Stripping Buffer, Cat. No. 62299 (20 mL) and Cat. No. 62300 (100 mL).
Note: Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membrane (Cat. No. 22860) only.
Will Restore and Restore Plus Western Blot Stripping Buffers remove biotin-bound avidin, streptavidin or NeutrAvidin Protein conjugates?
No. Restore and Restore Plus Buffers are not stringent enough to break the avidin-biotin interaction. A buffer capable of breaking this interaction would likely also damage the target protein, making it undetectable. However it may be possible to strip the complexed biotinylated primary antibody and streptavidin-HRP secondary antibody from the target protein on the membrane. Please also see Tech Tip: Strip and Reprobe Western Blots (https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/strip-reprobe-western-blots-tech-tip.pdf).
Will the Restore and Restore Plus Western Blot Stripping Buffers remove precipitating substrates?
No. The antibodies are removed but the substrate leaves a permanent precipitate on the membrane that cannot be removed. Restore and Restore Plus Buffers are designed for procedures using chemiluminescent substrates. Please note that this is not compatible with fluorescence supstrates as it will result in increased background. For fluorescent substrates please use our Restore Fluorescent Western Blot Stripping Buffer. Please also see Tech Tip: Strip and Reprobe Western Blots (https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/strip-reprobe-western-blots-tech-tip.pdf).