Titansphere TiO 填料

Titansphere TiO填料，是將(TiO2)做為骨架結(jié)構(gòu)的全新型HPLC用球狀凝膠填充劑. 是一款產(chǎn)業(yè)化了的凝膠色譜填料.以下為色譜填料。

Titansphere TiO填料是一種在堿條件下也同樣能夠使用的，與硅膠有著不同吸附特性的填充劑。其主要特點(diǎn)如下：
表面、形狀平整的球狀凝膠
表面積比較大
選擇性保留磷酸化合物
優(yōu)異的結(jié)構(gòu)異構(gòu)體分離能力
優(yōu)異的耐酸、耐堿性

磷酸化肽的精制和濃縮

細(xì)胞中的磷酸化蛋白質(zhì)，其存在量為微量且離子化效率低，因此質(zhì)譜（MS）檢測比較困難。但是，如果使用以（：TiO2）形成的均一多孔質(zhì)球狀粒子TitansphereTiO，

則可從蛋白質(zhì)酶切物中選擇性地濃縮精制磷酸化肽，使MS檢測變得容易。

操作方法全部為離心操作，幾乎沒有人為誤差。處理時間約40分鐘，可在短時間內(nèi)完成精制? 濃縮。

磷酸化肽選擇的精制濃縮珠

Titansphere TiO 填料是能夠選擇性地精制 濃縮磷酸化肽的高性能填料。使用擬南芥細(xì)胞提取物，與市售IMAC 進(jìn)行了比較，磷酸化肽的總峰面積約2.6 倍、肽鑒定數(shù)約1.8 倍，可檢測到大部分磷酸化肽。

Features

Spherically shaped porous particle “Titansphere TiO" (Titanium Dioxide ; TiO2) is a high-performing bulk material to enrich and purify phosphopeptides selectively.

• Efficient purification from HeLa Cell Lysate

The graph below shows the superior performance of Titansphere TiO using the HeLa Cell Lysate consisting mainly of non-phosphorylated peptides. Titansphere TiO shows exceptional selectivity – almost 90% of the bound peptides were phosphopeptides, and excellent capacity for total phosphopeptide binding. Whereas a competitive TiO product binds mainly non-phosphorylated peptides and a much lower total number of discrete phosphopeptides.

Sample: HeLa Cell Lysate,  Sample volume: 50 μg,  Titansphere TiO volume: 1 mg

• Comparison of Titansphere TiO and IMAC

The graph below shows the comparison of Titansphere TiO and IMAC enrichment using Arabidopsis cell extract. Titansphere TiO provides substantially higher total capacity and a much higher number of isolated phosphopeptides.

Sample: Arabidopsis Cell Extract,  Sample volume: 100 μg,  Titansphere TiO volume: 1 mg

• Example of how to make a Titanium Column
  ? Cut the Empore Disk C8 and put it at the tip of the 200ul pipette tip to be used as a filter.
  ? Stir and suspend the Titansphere TiO atCH3CN/H2O=80/20.v/v then load it to the tip.
    (Adjust the amounf of the beads depending on the purpose)
  ? Pressure the tip by a plastic syringe.

  

  
  

  

Specifications

訂貨信息

Identified Numbers of Phosphopeptides

Titansphere™ TiO Part Number

* New material for higher purification efficiency of Phosphopeptides

Empore Disk C8 Part Number

■參考文獻(xiàn)（混合タンパク質(zhì) トリプシン消化物からのオフライン精製）

1. M. R. Larsen, T. E. Thingholm, O. N. Jensen, P. Roepstorff, and T. J.D. Jorgensen,
“Highly selective enrichment of phosphorylated peptides from peptide mixtures using titanium dioxide microcolumns",
Molecular & Cellular Proteomics 2005; 4: 873-886.
＊混合タンパク質(zhì) 消化物からのリン酸化ペプチド 選択的濃縮條件の適化及び二酸化チタンカラムとIMACカラムとの性能比較

2. Jesper V. Olsen, Blagoy Blagoev, Florian Gnad, Boris Macek,
Chanchal Kumar, Peter Mortensen, and Matthias Mann.
“Global, In Vivo, and Site-Specific Phosphorylation Dynamics in Signaling Networks"
Cell 127, 635-648, November 3, 2006.
＊リン酸化を指標(biāo)とした細(xì)胞內(nèi)シグナル伝達(dá)の動的解析

3. Brend Bodenmiller, Lukas N Mueller, Murkus Mueller, Bruno Domon & Ruedi Aebersold.
“Reproducible isolation of distinct, overlapping segments of the phosphoproteome"
Nature Methods. Vol.4 No.3, 231-237(2007)
＊IMAC、チタニア等のショウジョウバエ培養(yǎng)細(xì)胞中リン酸化ペプチドの吸著特性比較

4. Naoyuki Sugiyama, Takeshi Masuda, Kosaku Shinoda, Akihiro Nakamura, Masaru Tomita, Yasushi Ishihama
“Phosphopeptide Enrichment by Aliphatic Hydroxy Acid-Modified Metal Oxide Chromatography for NanoLC-MS/MS in Proteomics Applications"
Molecular & Cellular Proteomics 2007;6:1103-1109.
＊チタニアを用いた、Hela細(xì)胞からのリン酸化ペプチド精製における選択的濃縮條件の適化

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