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.
Protocol
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.
Restrictions
For Research Use only
Reconstitution
Restore with 1 mL H2O (15 min, RT).
Buffer
PBS, 0.09 % Sodium Azide, PEG and Sucrose
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C
Storage Comment
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.
APG5 antibody, APG5-LIKE antibody, APG5L antibody, ASP antibody, hAPG5 antibody, ATG5 antibody, CG1643 antibody, DmAtg5 antibody, Dmel\\CG1643 antibody, atg5 antibody, 2010107M05Rik antibody, 3110067M24Rik antibody, AW319544 antibody, Apg5l antibody, Atg5l antibody, C88337 antibody, Paddy antibody, apg5l antibody, zgc:100934 antibody, ATATG5 antibody, AUTOPHAGY 5 antibody, MKP11.20 antibody, MKP11_20 antibody, autophagy related 5 antibody, Autophagy-related 5 antibody, ATG5 autophagy related 5 homolog (S. cerevisiae) antibody, autophagy related 5 L homeolog antibody, similar to S. cerevisiae ATG5 (YPL149W) which nucleates preautophagosome formation as a conjugate with Atg12 antibody, autophagy protein Apg5 family antibody, ATG5 antibody, Atg5 antibody, atg5 antibody, atg5.L antibody, APG5 antibody
Background
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