Minute™ 總蛋白萃取試劑盒(動物細胞和組織)
Minute™ Total Protein Extraction Kit for Animal Cultured Cells and Tissues
廠牌
產品簡介
Invent Minute™ 總蛋白萃取試劑盒（貨號 SD-001/SN-002）適用於動物培養細胞和動物組織樣本。採用 Minute™ 專利離心管柱與優化配方的細胞裂解液，僅需搭配一般 Vortex 與桌上型微量離心機，即可於 1–5 分鐘內完成總蛋白萃取實驗（萃取濃度 1–8 mg/ml，最少萃取體積可低至 20 µl）。相較於傳統 RIPA buffer 方式，本產品可萃取出更穩定全面的總蛋白質，提升實驗結果再現性。套組內附兩種細胞裂解液，研究人員可依下游實驗需求選擇萃取變性蛋白 (denatured proteins) 或保有功能的天然蛋白 (native proteins)。應用實驗包含 SDS-PAGE 電泳、Western Blot、ELISA、免疫沉澱 (immunoprecipitation)、酵素活性分析等。
Total proteins were extracted from mouse liver and splenocytes by Minute™ Total Protein Extraction Kit (SD-001) and RIPA buffer (Sigma, R0278).
Lane 1: Proteins extracted by Minute™ Kit.
Lane 2: Proteins extracted by RIPA buffer.
Lane 3: Proteins extracted from insoluble fraction of RIPA extraction by Minute™ Kit.
There is no insoluble fraction with Minute kit. Proteins extracted by RIPA buffer contain significantly fewer proteins in low molecular weight area (lane 2) but they are significantly increased in residual insoluble fraction (Lane 3). Based on these findings, it is obvious that the protein profile extracted by RIPA buffer is incomplete and altered.
(右) 與 RIPA buffer 方式相比，Minute™ Kit (SD-001) 萃取出的總蛋白更能真實維持原有的磷酸化蛋白質含量與狀態，不易造成結果誤判。
Image courtesy of G. Szanda, NIH/NIAAA/LPS.
Total proteins were extracted from zebrafish with RIPA buffer or Minute™ Kit (SD-001). Equal amount of proteins were loaded onto 4 lanes and probed with anti-GAPDH with the same dilution. The background is much cleaner with SD-001 extracted sample.
Image courtesy of Chengdu University of Chinese Medicine.
Isolated mouse brain cell suspension (20 μl) was lysed by either 200 μl Minute™ Kit (SD-001) Denaturing Cell Lysis Buffer or 200 μl RIPA buffer (Sigma, R0278) for 15 min. The cell lysates were centrifuged at 16,000 × g for 10 min. SD-001 shows complete cell lysis, no pellet is seen, while RIPA buffer shows incomplete lysis as evidenced by the presence of obvious insoluble pellet at the bottom of the tube (lower Right).
技術原理
離心管柱 (spin column) 形式的 DNA、RNA 萃取方式早已行之有年，美國 Invent Biotechnologies 公司則創新開發了可用於蛋白質萃取的離心管柱式分離技術 Minute™（美國專利號碼 US9580462B2）。Minute™ 離心管柱含有精密微孔構造的專利濾芯。進行總蛋白質萃取實驗時，先使用 Invent 公司自行研發優化的細胞裂解液 (cell lysis buffer) 將細胞膜和核膜打破，此時除了會釋放出蛋白質以外，原本包覆在細胞核膜內的 genomic DNA 也會隨之游離出來，使細胞裂解物 (total cell lysate) 變得非常黏稠。再將此黏稠的細胞裂解物樣本直接倒入 Minute™ 專利管柱中，置入桌上型微量離心機以離心力迫使細胞裂解物通過管柱。在極短的時間內，細胞碎片和超長鏈的 genomic DNA 即會被卡在管柱中，而蛋白質則可以迅速順利通過管柱，僅需 30 秒至 1 分鐘的離心時間，即可在下方收集管中收集到澄清的總蛋白質溶液。還可以依據搭配使用的細胞裂解液種類，選擇是要萃取保有原始構形與功能的天然蛋白或變性蛋白，以應用於下游分析實驗，例如 SDS-PAGE 電泳、Western Blot、ELISA、免疫沉澱 (immunoprecipitation)、酵素活性分析等。
Minute™ 專利技術不僅操作流程快捷簡易，並且僅需搭配一般 Vortex 與桌上型微量離心機，即可在極短的時間內取得高產量與高品質的總蛋白質，提升實驗效率與實驗結果再現性。
套組內容物
| Items | Quantity |
| Denaturing Cell Lysis Buffer | 25 ml |
| Native Cell Lysis Buffer | 25 ml |
| Protein Extraction Filter Cartridges | 50 units |
| Collection Tubes with Caps | 50 units |
| Plastic Rods | 2 units |
儲存條件
於室溫下儲存可穩定保存 24 個月。