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
| Description | Titansphere TiO |
|---|---|
| Particle Size | 5 μm, 10 μm |
| Particle Shape | Spherical |
| Adsorption Spot | Titanium Dioxide Crystal |
| Pore Size | 100 A (10 μm) |
| pH Range | 2 - 12 |
| Gravity | 1.74 |
訂貨信息
Identified Numbers of Phosphopeptides
| Phosphorylate | Non - Phosphorylate | |
|---|---|---|
| Titansphere TiO | 846 | 198 |
| IMAC | 474 | 379 |
Titansphere™ TiO Part Number
| Description | Volume | Cat.No. |
|---|---|---|
| Titansphere TiO 5 μm | 500 mg | 5020-75000 |
| Titansphere TiO 10 μm | 500 mg | 5020-75010 |
| * Titansphere Phos-TiO Bulk 10 um | 500 mg | 5010-21315 |
* New material for higher purification efficiency of Phosphopeptides
Empore Disk C8 Part Number
| Description | Quantity | Cat.No. |
|---|---|---|
| Empore Disk C8, 47 mmDIA, 0.5 mm Thickness | 20 pcs | 5010-30002 |
| Empore Disk C8, 90 mmDIA, 0.5 mm Thickness | 10 pcs | 5010-30003 |
■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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3M Empore disk C8 固相萃取膜片
Resprep-C18 and Resprep-C8 SPE Disks膜片
