Western Blot: 0.5 μg/mL for HRPO/ECL detection. Recommended buffer: Casein/Tween 20 based blocking and blot incubation buffer. Use cell lysate from EGF-treated SKOV-3 cells as positive control (please see Protocolsbelow for details). Immunocytochemistry. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
实验流程
POSITIVE CONTROL CELL LYSATE: SKOV-3 EGF treatedFormat: Lyophilized cell lysate from SKOV-3 cells. Serum starved cells were treated for 15min with EGF. Reconstitute by addition of 200 µl H2O. After complete solubilization add 200 µl 2xSDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Applications: The positive control cell lysate is recommended for immunoblot applications. 20 µl of positive control cell lysate correspond to ca. 20. 000 cells. Use 20 µl / lane (mini gel) for HRPO/ECL detection of the target proteins. Please note: The lyophilized cell lysates contain SDS and are not recommended for applications withnative proteins such as immunoprecipitation. Storage: In aliquots at -20°C. Avoid repeated freezing and thawing.
限制
仅限研究用
溶解方式
Restore with 1 mL H2O (15 min, RT).
缓冲液
1 mLPBS containing 0.09 % Sodium Azide/PEG and Sucrose.
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
-20 °C/-80 °C
储存方法
Store lyophilized (preferably in a desiccator) at -20 °C and reconstituted (aliquote andfreeze in liquid nitrogen) at -80 °C. Avoid repeated freezing and thawing. Shelf life: one year from despatch. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 4 °C up to 3 months.
ErbB2 is a member of the EGFR/erbB-receptor tyrosine kinase family. Dysregulation of erbB2 and/or activation of downstream signaling pathways has been implicated in many human cancers. ErbB2 is activated upon ligand dependent heterodimerization with EGFR or erbB4. ErbB2 homodimers are not favored due to the lack of an erbB2 specific extracellular ligand. Heterodimerization with EGFR or erbB4 leads to activation of the intrinsic tyrosine kinase activity of EGFR or erbB4 resulting in phosphorylation of multiple tyrosine residues within the erbB2 intracellular domain: Tyr 1023, Tyr 1112, Tyr 1139, Tyr 1196, Tyr 1222, and Tyr 1248.Transphosphorylation via src family kinases leads to phosphorylation of Tyr 877, via PKC of Thr 686, via CamKinase2 of Ser 1113. Phosphorylation of Thr 686 and Ser 1113 interferes with erbB2 endocytosis and degradation.Synonyms: HER-2, MNL19, NEU, NEU proto-oncogene, NGL, Receptor tyrosine-protein kinase erbB-2, Tyrosine kinase-type cell surface receptor HER2, c-erbB-2, p185erbB2