Immunofluorescent analysis of Beclin-1 (green) HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with a Beclin-1 polyclonal antibody (Product # PA5-20171) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A 16 amino acid peptide from near the carboxy terminus of human Beclin-1.|
|Purification||Antigen affinity chromatography|
|Contains||0.02% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||0.5-2 µg/ml|
* 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.
A suggested positive control is A431 cell lysate.
PA5-20171 can be used with blocking peptide PEP-0288.
Autophagy, the process of bulk degradation of cellular proteins through an autophagosomic-lysosomal pathway is important for normal growth control and may be defective in tumor cells. Beclin-1, a coiled-coil Bcl-2-interacting protein homologous to the yeast autophagy gene apg6, is a mammalian autophagy gene that can inhibit tumorigenesis and is expressed at reduced levels in human breast carcinoma, suggesting that defects in autophagy proteins may contribute to the development or progression of tumors. Bcl-2 can bind to Beclin-1 and inhibit Beclin-1-dependent autophagy in yeast and mammalian cells, suggesting that Bcl-2 functions as an anti-autophagy protein as well as an anti-apoptotic protein, which helps maintain autophagy at levels that are more compatible with cell survival rather than cell death.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.