Anti phospho bad ser136

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-phospho-Bad (Ser136) is a lab equipment product that detects the phosphorylation of the Bad protein at serine 136. This product is used to study signaling pathways and cellular processes involving the regulation of the Bad protein.

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Close spelling variants of this product

Anti-Bad (42 citations) Phospho-Bad (10 citations) Anti-phospho-Bad (8 citations) Phospho-BAD-ser136 (2 citations)

3 protocols using anti phospho bad ser136

Mitomycin C and Rapamycin Protocol

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Mitomycin C was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rapamycin and protease inhibitor cocktail were obtained from Sigma-Aldrich (St Louis, MO, USA). S6K1WT-HA, S6K1E389DeltaCT-HA and S6K1F5A were purchased from Addgene (Cambridge, MA, USA). Anti-HA, anti-phospho-Akt, anti-Akt, anti-CEA, anti-phospho-JNK, anti-JNK, anti-Bak, anti-caspase 8, anti-caspase 9, anti-caspase 3, anti-phospho-p70 S6 kinase (Thr389), anti-p70 S6 kinase, anti-phospho-Bad (Ser136), anti-Bad, anti-COX-IV and anti-PARP antibody were from Cell Signaling (Danvers, MA, USA). Anti-actin antibody was from Sigma-Aldrich. Anti-cytochrome c antibody was from BD PharMingen (San Jose, CA, USA). Anti-Ki67 was purchased from Dako (Carpinteria, CA, USA).

Song X., Dilly A.K., Kim S.Y., Choudry H.A, & Lee Y.J. (2014). Rapamycin-enhanced mitomycin C-induced apoptotic death is mediated through the S6K1–Bad–Bak pathway in peritoneal carcinomatosis. Cell Death & Disease, 5(6), e1281-.

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BDNF Signaling Pathway Antibody Detection

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Following primary antibodies were used in this study: rabbit polyclonal anti-BDNF (1:1000, #ab226843, Abcam), rabbit monoclonal anti-TrkB (1:1000, #4603, Cell Signaling Technology), anti-p75^NTR (1:1000, #4201, Cell Signaling Technology), anti-phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (1:1000, #4621, Cell Signaling Technology), anti-p44/42 MAPK (Erk1/2) (1:1000, #4695, Cell Signaling Technology), anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (1:1000, #4377, Cell Signaling Technology), anti-Akt (pan) (1:1000, #4691, Cell Signaling Technology), anti-phospho-Akt (Ser473) (1:1000, #4058 Cell Signaling Technology), anti-BAD (1:1000, #9239, Cell Signaling Technology), anti-phospho-BAD (Ser136) (1:1000, #4366, Cell Signaling Technology), anti-phospho-GSK-3β (Ser9) (1:1000, #9323, Cell Signaling Technology), rabbit polyclonal anti-phospho-TrkB (Tyr816) (1:1000, #NBP1-03499SS, Novus Biologicals), anti-phospho-TrkB (Tyr515) (1:1000, # PA5-36695, Thermo Fisher Scientific), anti-Caspase-9 Antibody (1:1000, #9502, Cell Signaling Technology), mouse monoclonal anti-α-Tubulin (1:1000, #2125, Cell Signaling Technology). Secondary antibodies conjugated to HRP (used at 1:12,000 dilution) were from Cell Signaling Technology (anti-mouse IgG, #7076; anti-rabbit IgG, #7074).

Zainullina L.F., Vakhitova Y.V., Lusta A.Y., Gudasheva T.A, & Seredenin S.B. (2021). Dimeric mimetic of BDNF loop 4 promotes survival of serum-deprived cell through TrkB-dependent apoptosis suppression. Scientific Reports, 11, 7781.

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Signaling Pathways Activated by Growth Factor Stimulation

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Cells were plated at a density of 5×10⁵ per well. Cells were allowed to attach overnight and were serum starved for 24 hours before treatment. Cells were treated with vehicle or inhibitors one hour prior to 5% FBS stimulation for 24 hours. For Bad signaling, 50ng/mL EGF for 4 hours was used as a stimulus. Following treatment, cells were lysed and protein content was determined by a Bradford assay (Bio-Rad). 30µg of protein was loaded on a 4–15% SDS-PAGE gradient gel (Bio-Rad). The contents of the gel were transferred to a membrane and then probed with various antibodies: anti-phospho-ERK1/2, anti-total ERK1/2, anti-total ERK5, anti-phospho-ERK5, anti-phospho-Akt (Ser 473), anti-Akt, anti-pS6 (Ser 240/244), anti-p21, anti-cMYC, anti-Bad, and anti-phospho-Bad (Ser136) (1:1000, Cell Signaling). Anti-GAPDH (Millipore) was used as a loading control. The binding of antibody to antigen was detected by incubating membranes with secondary antibodies and scanning on an Odyssey Infrared Imager (LICOR biosciences). Blots were analyzed using ImageStudioLite by a blinded observer (LICOR biosciences). Phospho-Bad Ser112 was quantified with PathScan ELISA (Cell Signaling; 7182C).

Wright T.D., Raybuck C., Bhatt A., Monlish D., Chakrabarty S., Wendekier K., Gartland N., Gupta M., Burow M.E., Flaherty P.T, & Cavanaugh J.E. (2019). Pharmacological inhibition of the MEK5/ERK5 and PI3K/Akt signaling pathways synergistically reduces viability in triple-negative breast cancer. Journal of cellular biochemistry, 121(2), 1156-1168.

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