Western Blotting (WB), ELISA, Immunoprecipitation (IP), Indirect Immunofluorescence Assay (IFA)
特异性
Binding of EGF to the extracellular domain of the EGF-receptor results in receptor dimerization and autophosphorylation on tyrosine residues, Y1173 being the major autophosphorylation site. Clone 9H2 specifically interacts with the 1170 - NAEpYLRV motif corresponding to the major autophosphorylation site of human EGFR. The antibody does not interact with the non-phosphorylated EGFR nor with unrelated Tyrosine-phosphorylated proteins. This phosphorylation site-specific monoclonal antibody allows the detection and quantification of activated EGFR in crude cell extracts in Immunoblot, ELISA and Biosensor assay formats without the use of P32.
交叉反应 (详细)
Reacts with Human, mouse, and dog.
产品特性
Cell lysate of pervanadate-treated HepG2 cells, Cell culture
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: 0.5 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) 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 K67902M for 1 hour at RT and developed by ECL (exp. time: 30 sec). Immunoprecipitation: 1-10ug per 106 vanadate treated A431 cells. Immunocytochemistry: 1-10 µg/mL ELISA: 0.05 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.