- SynonymEGF,URG,HOMG4
- SourceHuman EGF, Mouse IgG2a Fc Tag (EGF-H525b) is expressed from human 293 cells (HEK293). It contains AA Asn 971 - Arg 1023 (Accession # P01133-1).Predicted N-terminus: GluRequest for sequence
- Molecular Characterization
This protein carries a mouse IgG2a Fc tag at the N-terminus.
The protein has a calculated MW of 33.1 kDa. The protein migrates as 35 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
- EndotoxinLess than 0.1 EU per μg by the LAL method.
- Purity
>95% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in TriswithGlycine,ArginineandNaCl,pH7.5. 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 EGF, Mouse IgG2a Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Immobilized Human EGF R, Fc Tag (Cat. No. EGR-H5252) at 5 μg/mL (100 μL/well)can bind Human EGF, Mouse IgG2a Fc Tag (Cat. No. EGF-H525b) with a linear range of 0.3-10 ng/mL (QC tested).
- Citations
A Wearable and Deformable Graphene-Based Affinity Nanosensor for Monitoring of Cytokines in Biofluids
Authors: Wang Z, Hao Z, Yu S, et al.
Journal: NANOMATERIALS-BASEL 2020
Application: biosensor
Request for Full-text
- BackgroundHuman epidermal growth factor (EGF) is also known as HOMG4 and URG,and is a growth factor that plays an important role in the regulation of cell growth, proliferation, and differentiation by binding to its receptor EGFR. Epidermal growth factor can be found in human platelets, macrophages, urine, saliva, milk, and plasma. EGF is the founding member of the EGF-family of proteins. Members of this protein family have highly similar structural and functional characteristics. All family members contain one or more repeats of the conserved amino acid sequence. The biological effects of salivary EGF include healing of oral and gastroesophageal ulcers, inhibition of gastric acid secretion, stimulation of DNA synthesis as well as mucosal protection from intraluminal injurious factors such as gastric acid, bile acids, pepsin, and trypsin and to physical, chemical and bacterial agents. Because of the increased risk of cancer by EGF, inhibiting it decreases cancer risk.
- References
(1)Barnham KJ,et al. 1998, Protein Science 7 (8): 1738–49.
(2)Cotran, Ramzi S.,et al. 2005, Robbins and Cotran pathologic basis of disease.
(3)Dreux AC,et al. 2006, Atherosclerosis 186 (1): 38–53.
(4)Venturi S,et al.2009, Nutrition and Health 20 (2): 119–134.
(5)Herbst RS., 2004, Int J Radiat Oncol Biol Phys., 59(2 Suppl): 21-6.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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对于瞬时转染的检测：如果有报告基因，就通过报告基因的表达看转染成功与否；没有报告基因的话，就在24～96h内收获细胞通过RT-PCR和WB来鉴定。
1.将外源基因插入慢病毒载体
2.将构建完成的载体与慢病毒包装质粒混合，共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选，如果没有，可以用无限稀释法
6.获得稳转株
一种是脂质体转染后,单克隆筛选稳定细胞株.另外一种是应用逆转录病毒,慢病毒转染,筛选稳定细胞株.
脂质体转染：在转染24小时后,消化细胞并计数.将细胞种到96孔板,保证每个孔2-3个细胞,这样才能得到单克隆.待细胞贴壁后,加入抗生素筛选.筛选时间和浓度视细胞而定.一般G418一个星期作用,嘌呤霉素2-3天.
脂质体法筛单克隆时间较长,且效率低,大概只有1%.
病毒转染：先要用包装细胞,一般为293细胞,包装出病毒,再用病毒转染目的细胞.包装病毒视不同类型的病毒而定,一般要3-5天的时间.包装好的病毒要测滴度,根据滴度决定转染目的细胞的病毒量.转染目的细胞1-2天后加抗生素筛选得到稳定细胞株.
病毒转染得到稳定细胞株的效率高,只是步骤繁琐.
推荐了解“主细胞库”“工作细胞库”这两个名词。
① 在构建载体时，目的基因直接整合到细胞染色体组上，最好不要通过先瞬转在筛选稳定细胞株的这种方法，因为转染效率没有保证
② 高表达载体的构建，哺乳动物表达量一直是它自身的缺点，最好根据高表达载体定向的驯化细胞，提高蛋白表达量
③ 细胞的选择，筛选稳定细胞株我们常用的细胞是CHO，中国仓鼠卵巢细胞，由于CHO具有诸多的优点因此适合用于筛选稳定细胞株，而HEK293细胞则常用于瞬时转染
④ 后期的筛选，双抗预防污染，筛选细胞的时候抗生素浓度一定要做预实验，而且转染的时候不能有抗生素，关于细胞转染 稳定细胞系构建的相关理论
