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Titansphere TiO 二氧化鈦填料 (GL Sciences )

詳細(xì)介紹

Titansphere TiO 二氧化鈦填料


 

 

Titansphere TiO填料,是將二氧化鈦(TiO2)做為骨架結(jié)構(gòu)的全新型HPLC用球狀二氧化鈦凝膠填充劑. 是一款產(chǎn)業(yè)化了的二氧化鈦凝膠色譜填料.以下為二氧化鈦色譜填料。

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、形狀平整的球狀二氧化鈦凝膠
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(cè)比較困難。但是,如果使用以二氧化鈦(二氧化鈦: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(cè)變得容易。

操作方法全部為離心操作,幾乎沒(méi)有人為誤差。處理時(shí)間約40分鐘,可在短時(shí)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(cè)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

Application-Identified Peptides

  • 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

Application-Comparison with IMAC

  • 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.


    Titanium Column

Specifications

DescriptionTitansphere TiO
Particle Size5 μm, 10 μm
Particle ShapeSpherical
Adsorption SpotTitanium Dioxide Crystal
Pore Size100 A (10 μm)
pH Range2 - 12
Gravity1.74


訂貨信息


Identified Numbers of Phosphopeptides

 PhosphorylateNon - Phosphorylate
Titansphere TiO846198
IMAC474379

Titansphere TiO Part Number

DescriptionVolumeCat.No.
Titansphere TiO 5 μm500 mg5020-75000
Titansphere TiO 10 μm500 mg5020-75010
* Titansphere Phos-TiO Bulk 10 um500 mg5010-21315

* New material for higher purification efficiency of Phosphopeptides

Empore Disk C8 Part Number

DescriptionQuantityCat.No.
Empore Disk C8, 47 mmDIA, 0.5 mm Thickness20 pcs5010-30002
Empore Disk C8, 90 mmDIA, 0.5 mm Thickness10 pcs5010-30003


■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(dòng)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


3.jpg

Resprep-C18 and Resprep-C8 SPE Disks膜片


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