Western blot analysis was performed on whole cell extracts (20 ug lysate) of HeLa (Lane 1), HeLa treated for 20 minutes with 200 nM of PMA (Lane 2), A549 (lane 3), Serum Starved A549 (lane 4) and A549 Serum Starved for overnight followed by Serum Released (lane 5). The blots were probed with Anti-Ribosomal Protein S6 [pS235] / [pS236] Rabbit Polyclonal Antibody (Product# 44922G, 0.5-2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 31 kDa band corresponding to Ribosomal Protein S6 [pS235] / [pS236] were observed across PMA treated and Serum Starved followed by Serum Released cell lines tested. 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 following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human RPS6 that contains serines 235 and 236. The sequence is conserved in mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|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.
|Western Blot (WB)||See 1 publications below|
Ribosomal protein 6 (RPS6) is a component of the 40S ribosomal subunit belonging to S6E family of ribosomal proteins. RPS6 is a key substrate for kinases, and is phosphorylated by growth factors and mitogens during in cell growth and cell division. Phosporylation in RPS6 is well regulated, and the different phosphorylation sites are highly conserved. Major phosphorylation sites of RPS6 include Ser 235, 236, 240, and 244.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Stimulation of autophagy by rapamycin protects neurons from remote degeneration after acute focal brain damage.
44-922G was used in western blot to investigate the role of autophagy in acute brain damage.
|Viscomi MT,D'Amelio M,Cavallucci V,Latini L,Bisicchia E,Nazio F,Fanelli F,Maccarrone M,Moreno S,Cecconi F,Molinari M||Autophagy (8:222)||2012|