
- 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.
ebiomall.com






>
>
>
>
>
>
>
>
>
>
>
>
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。
不建议你用MDCK细胞,非常难转染。
GFP发出绿色荧光的原理是Ca离子进入GFP的beta-barrel结构中引起的特定能级,因此只要这个结构仍然保持着,就可以发出荧光。
由于GFP的beta-barrel结构非常稳定,一些版本的GFP蛋白(如EGFP)甚至能抵抗94C的高温几分钟而不完全变性,因此想在溶液状态下去掉GFP的荧光是很难的,一般需要用光漂白法。
基于其非常稳定的结构,即便细胞被固定了,仍然会有一部分的GFP蛋白保持其构象而发出荧光。此时荧光可能较弱。在荧光显微镜下是有可能看得到的。
我刚开始做转染,悬浮细胞,分别做过表达和敲减,看了很多文献,大都没有提及转染后是用转染的这同一批细胞同时做pcr,wb,cck8,凋亡,细胞周期;还是说这次转染只做pcr或wb,再转染一次做cck8或细胞周期。剩下的功能试验均同前,转染一次做一次?我养的是悬浮细胞,转染后做cck8这些功能试验前需要离心换液吗?跪谢解答!
可以用移液器将细胞吸出来并高速离心,沉淀重悬于PBS中洗涤,接着就可以裂解提取蛋白了。可以用超声,酶解等等,裂解后离心收集上清。
其次要看下你选择单位的规模如何,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。
实验室一直都是用日常型质粒抽提试剂盒,转染细胞没问题。
我觉得只要是注意以下2点就可以了:
1,注意大肠杆菌(Escherichia coli)本身的污染,收集菌体沉淀时防止菌液散落,经常用75%的乙醇擦拭手套。
2,最后洗脱时最好使用无内毒素的水,我们是用注射用水的。
无论是小提还是大提我们都是用的日常型的,并没有刻意用转染级的,因为转染量大,去内毒素的操作太麻烦,损失太大。
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。

