
- SynonymSPP1,BNSP,OPN,Uropontin,Nephropontin,Osteopontin,BSP-1,ETA-1,BSPI
- SourceHuman Osteopontin, His Tag (OPN-H5227) is expressed from human 293 cells (HEK293). It contains AA Ile 17 - Asn 300 (Accession # AAH07016).Predicted N-terminus: Ile 17Request for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 33.0 kDa. The protein migrates as 45-55 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>80% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.

Human Osteopontin, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 80%.
- BackgroundOsteopontin (OPN) is also known as Secreted phosphoprotein 1 (SPP1), Bone sialoprotein 1, Nephropontin, Urinary stone protein, Uropontin, BNSP, which belongs to the osteopontin family. OPN / SPP1 is a highly negatively charged, extracellular matrix protein that lacks an extensive secondary structure. Full length OPN (OPN-FL) can be modified by thrombin cleavage, which exposes a cryptic sequence, SVVYGLR on the cleaved form of the protein known as OPN-R. Osteopontin / SPP-1 is biosynthesized by a variety of tissue types. OPN is the ligand for integrin alpha-V/beta-3. OPN / SPP1 binds tightly to hydroxyapatite and appears to form an integral part of the mineralized matrix. OPN / SPP1 probably important to cell-matrix interaction. OPN / SPP1 acts as a cytokine involved in enhancing production of interferon-gamma and interleukin-12 and reducing production of interleukin-10 and is essential in the pathway that leads to type I immunity.
- References
- (1)Wang KX., et al., 2008, Cytokine Growth Factor Rev. 19 (5-6): 333–45.
- (2)Laffón A., et al., 1991, J. Clin. Invest. 88 (2): 546–52.
- (3)Christensen B., et al., 2005, Biochem. J. 390:285-292.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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细胞状态与密度;转染试剂的类型、用量;DNA的品质、用量;转染复合物的品质、作用细胞的时间长度。
其次要看下你选择单位的规模如何,做的比较好的,还是上海这边的,你可以看下基尔顿生物,原代细胞培养,动物造模,整体课题外包。
我转的是7901、7901/DDP两种细胞,前者7901细胞很容易就转上,并且转后,状态良好,可是7901/DDP一转就死,我用的是吉玛慢病毒,转24小时后换液,刚开始一两天,没有异常,但后来细胞慢慢就死了,并且不是漂浮的,很多是贴着壁死,像是瓦解了一样
这是未转时细胞的样子
这是细胞转后,死亡的样子
并且即使是有些细胞未死,细胞后来也变得很脏,感觉有很破碎的细胞碎片
本人实验小白,**园子里大神指点,急,实在不知道怎么回事
转染分2种,一种是瞬时转染,即转染后让细胞表达目的蛋白后即提取蛋白,提一次蛋白,转染一次,这种方式一般不传代;
另一种转染为稳定转染,转染后加入一定选择压力进行筛选,没有转染的细胞不能存活,只留下转染的细胞,这种情况下可以筛选单个转染细胞,构建稳定表达某一特定蛋白或基因的细胞系。
各位版友求助,
我使用Hek293构建转染模型,瞬转5质粒,用lipo2000做转染体系。
转染48h,发现荧光较强的细胞都在爬片的边缘,比例十分少。是因为我添加试剂的手法不对吗。
同时也发现加入转染体系后细胞状态特别差。想问一下用lipo2000时可以用无双抗的10%FBSDMEM吗。我转染前6h现在用的是纯DMEM,不含FBS。
请各位大神帮忙

