Immunobotting (Western Blot): 0.5 μg/mL for HRPO/ECL detection. Recommended blocking buffer: Casein/Tween 20 based blocking buffer and blotincubation buffer. Immunocytochemistry: 0.5 μg/mL. Flow Cytometry. Included Postitive Control: Cell lysate from untreated SH-SY5Y cells (See Protocols for moredetails). 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 from untreated SH-SY5Y cells. Formulation: Lyophilized cell lysate from Serum starved SH-SY5Y cells. Stability: 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. Application: The Positive Control lysate is recommended for Immunoblot applications. 20µl of PositiveControl correspond to ca. 20. 000 cells. Use 20µl/lane (mini gel) for HRPO/ECL detection of the target proteins. Storage: Aliquote and store frozen. Avoid repeated freeze/thaw cycles. Shelf life: one year from despatch. The Lyophilized cell lysates contain SDS and are not recommended for applications withnative proteins such as Immunoprecipitation.
限制
仅限研究用
溶解方式
Restore with 1 mL H2O (15 min, RT).
缓冲液
PBS, 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
储存方法
Store lyophilized (preferably in a desiccator) at -20 °C and reconstituted (aliquote andfreeze in liquid nitrogen) at -20 °C to -80 °C. Avoid repeated freezing and thawing. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 2-8 °C up to 3 months. Shelf life: one year from despatch.
Autophagy as a response to cellular stress and starvation is an alternative process of proteasomal degradation for some long-lived proteins and organelles. Alterations in the autophagic-lysosomal compartment have been linked to neuronal death in many neurodegenerative disorders as well as in transmissible neuronal pathologies (prion diseases). The gene product of autophagy-related gene 5 (ATG5) is required for autophagosome-formation. ATG5 also enhances the susceptibility towards apoptotic stimuli. Like Bcl-2, ATG5 exhibits a dual function by modulating both autophagy and apoptosis.Synonyms: APG5-like, ASP, Apoptosis-specific protein, Autophagy protein 5