Titansphere TiO 二氧化钛填料

Titansphere TiO填料，是将二氧化钛(TiO2)做为骨架结构的全新型HPLC用球状二氧化钛凝胶填充剂. 是一款产业化了的二氧化钛凝胶色谱填料.以下为二氧化钛色谱填料。

Titansphere TiO填料是一种在碱条件下也同样能够使用的，与硅胶有着不同吸附特性的填充剂。其主要特点如下：
表面、形状平整的球状二氧化钛凝胶
表面积比较大
选择性保留磷酸化合物
优异的结构异构体分离能力
优异的耐酸、耐碱性

磷酸化肽的精制和浓缩

细胞中的磷酸化蛋白质，其存在量为微量且离子化效率低，因此质谱（MS）检测比较困难。但是，如果使用以二氧化钛（二氧化钛：TiO2）形成的均一多孔质球状粒子TitansphereTiO，

则可从蛋白质酶切物中选择性地浓缩精制磷酸化肽，使MS检测变得容易。

操作方法全部为离心操作，几乎没有人为误差。处理时间约40分钟，可在短时间内完成精制? 浓缩。

磷酸化肽选择的精制浓缩珠

Titansphere TiO 填料是能够选择性地精制 浓缩磷酸化肽的高性能填料。使用拟南芥细胞提取物，与市售IMAC 进行了比较，磷酸化肽的总峰面积约2.6 倍、肽鉴定数约1.8 倍，可检测到大部分磷酸化肽。

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

订货信息

Identified Numbers of Phosphopeptides

Titansphere™ TiO Part Number

* New material for higher purification efficiency of Phosphopeptides

Empore Disk C8 Part Number

■参考文献（混合タンパク質 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 消化物からのリン酸化ペプチド 選択的濃縮条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.
＊リン酸化を指標とした細胞内シグナル伝達の動的解析

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細胞中リン酸化ペプチドの吸着特性比較

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細胞からのリン酸化ペプチド精製における選択的濃縮条件の適化

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3M Empore disk C8 固相萃取膜片

Resprep-C18 and Resprep-C8 SPE Disks膜片