Western blot analysis was performed on whole cell extracts (30 ug lysate) of Jurkat (Lane 1), Jurkat treated for overnight with 3 uM of Staurosporine (Lane 2), HeLa (Lane 3), HeLa treated for overnight with 3 uM of Staurosporine (lane 4), A549 (lane 5), PC-3 (lane 6) and MDA-MB-231 (lane 7). The blots were probed with Anti-Caspase 8 Mouse Monoclonal Antibody (Product# MA511558, 2-4 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 626520, 1:4000 dilution). A 55 kDa band corresponding to Caspase 8 was observed across cell lines tested except Staurosporine treated lysates. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product# SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant full length human Caspase 8 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||2 µg/ml|
|Western Blot (WB)||2-4 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
MA5-11558 targets Caspase 8 in IP and WB applications and shows reactivity with Human samples.
The MA5-11558 immunogen is recombinant full length human Caspase 8 protein.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Autocrine production of interleukin-4 and interleukin-10 is required for survival and growth of thyroid cancer cells.
MA5-11558 was used in immunocytochemistry to study the role of autocrine IL-4 and IL-10 in the survival and growth of thyroid cancer cells
|Todaro M,Zerilli M,Ricci-Vitiani L,Bini M,Perez Alea M,Maria Florena A,Miceli L,Condorelli G,Bonventre S,Di Gesù G,De Maria R,Stassi G||Cancer research (66:1491)||2006|