Binding of EGF leads to autophosphorylation of the EGF Receptor at 5 tyrosine residues. In addition, EGF Receptor is phosphorylated by members of the src kinase family at different tyrosine residues, phosphotyrosine 845 in the activation loop being the major site (tyrosine 845 is not an autophosphorylation site). EGFR acquires its full kinase activity only upon autophosphorylation and phosphorylation at tyrosine 845. Clone 12A3 allows the detection of EGFR activation after interaction with src kinases. Specifically recognizes EGFR phosphorylated at Tyrosine 845. The antibody does not interact with the non-phosphorylated EGFR nor with unrelated Tyrosine-phosphorylated proteins.
交叉反应 (详细)
Reacts with Human, mouse and rat.
产品特性
Cell lysate of 2 x 10^6 EGF treated HepG2 cells.
纯化方法
Thiophilic adsorption and size exclusion Chromatography. Cell culture
EGFR
适用: 人
WB, IHC, IP
宿主: 兔
Polyclonal
unconjugated
应用备注
Immunoblotting: 0.1 µg/mL for HRP/ECL detection. Recommended blocking buffer CPPT: 0.5 % (w/v) casein, 5 % (w/v) PEG 4000, 5 % (w/v) Polyvinylpyrrolidone (PVP), 0.5 % Tween 20, 10mM Tris/HCl, pH 7.4, 150mM Sodium Chloride. Phosphospecificity Whole cell extracts of control (co), EGF stimulated (EGF) or pervanadate treated (VH) HepG2 and A549 tumor cells were applied to SDS-PAGE (20.000 cells per lane) and transferred to a PVDF membrane. The blot was probed with 0.5- g/mL ABIN235076 for 1 hour at RT and developed by ECL (exp. time: 30 sec). ELISA: 0.1 µg/mL Each laboratory should determine an optimum working titer for use in its particular application. Other applications have not been tested but use in such assays should not necessarily be excluded.
限制
仅限研究用
状态
Lyophilized
溶解方式
Reconstitute with 1 mL water for 15 minutes at room temperature.
缓冲液
Lyophilized from PBS, PEG and Sucrose, 0.09 % Sodium azide
储存液
Sodium azide
注意事项
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
储存条件
4 °C/-20 °C/-80 °C
储存方法
Store lyophilized product at -20 °C. After reconstitution, aliquot and store at -80 °C. Thawed aliquots may be stored at 2-0 °C for up to 3 months.