Immunofluorescence analysis of Caspase-8 [pS387] was done by treating HeLa cells with TNF-alpha at 20 ng/mL for 20 minutes. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ Caspase-8 [pS387] Recombinant Rabbit Oligoclonal Antibody (710535) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). F-actin (Panel b: red) was stained with Alexa Fluor® 594 Phalloidin (A12381) and nuclei (Panel c: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). Panel d is a merged image showing nuclear localization respectively and e shows competition with the phospho- Caspase 8 [pS387] peptide. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphorylation of the Caspase 8 at the Ser-387 residue|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:5000|
* 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
Caspase-8 binds to the death effector domain (DED) of FADD through an analogous DED domain present in tandem in the proform of the caspase-8 protein. Recruitment of caspase-8 to the Fas receptor results in oligomerization of the caspase-8 protein, which in turn drives its autoactivation through "self-cleavage". Activated caspase-8 then activates other downstream caspases including caspase-9, thereby commiting the cell to undergo apoptosis. This antibody also detects the processed forms (42/44kDa, 25kDA, and 14kDa) of caspase-8.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
PCTAIRE1-knockdown sensitizes cancer cells to TNF family cytokines.
710535 was used in western blot to determine the role of PCTAIRE1 in apoptosis.
|Yanagi T,Shi R,Aza-Blanc P,Reed JC,Matsuzawa S||PloS one (10:null)||2015|