Immunofluorescent analysis of ATG9 (green) accumulation in HeLa cells in response to serum starvation (0.1% FCS, 16 hours). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with an ATG9 monoclonal antibody (Product # MA1-149) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-Armenian Hamster IgG secondary antibody. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification. Accumulation and redistribution (red arrows) of cytoplasmic ATG9 in response to starvation-induced autophagosome assembly is shown (right panel).
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Armenian hamster / IgG|
|Immunogen||peptide sequence HPEPVPEEGSEDELPPQVHK of human ATG9A C-terminus|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-149 was produced in Armenian hamster and detects ATG9 in human, rat and mouse samples. MA1-149 has been successfully used in Western Blot, Immunofluorescence, and ELISA procedures. Western Blot analysis with MA1-149 shows the detection of a double band at ~85-95 kDa in human ATG9 overerexpression lysates. MA1-149 also detects additional unknown bands at ~40 and ~60 kDa. In Immunofluorescence applications, MA1-149 shows accumulation and redestribution of ATG9 in response to starvation-induced autophagosome assembly.
Autophagy is an important process associated with infectious disease, neurodegeneration, cancer and the maintenance of cellular homeostasis. ATG9 is an essential component of the autophagy machinery and is found on autophagosomes. ATG9 forms a cycling complex with ATG2 and ATG18, which is thought to constantly deliver lipids from source to growing autophagosomes during early autophagosome assembly. ATG9 is a multi-spanning membrane protein and cycles between a juxta-nuclear trans-Golgi network compartment and late endosomes. Nutrient starvation induces accumulation on autophagosomes. The detection of this protein provides important information about cell health and survival.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.