Akt sc 8312

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Akt(sc-8312) is an antibody that recognizes the Akt protein. Akt is a serine/threonine-specific protein kinase that plays a key role in multiple cellular processes, including cell proliferation, survival, and metabolism. The Akt(sc-8312) antibody can be used to detect and analyze Akt expression and activation in various experimental systems.

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6 protocols using akt sc 8312

Phosphorylation Signaling Pathway Antibodies

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Anti-Flag was obtained from Abigent. Antibodies against C-Myc(sc-289) and Akt(sc-8312) were from Santa Cruz Biotechnology Inc. Antibodies against p-Akt^Ser473(4060), p-Akt^Thr308(13038), p-p70S6K^Thr389(9234), p-4EBP1^Thr37/46(2855) and HA(3724) were obtained from Cell Signaling Technology. Anti-IRS4 (ab52622) and CK1γ2 (GTX33123) were purchased from Abcam and GeneTex, respectively. Antibodies against GAPDH (13937-1-AP) and β-Tublin(10094-1-AP) were from Proteintech. Anti-p-IRS4-S859, an antibody specific for the phosphorylation on Ser859 of IRS4, was generated by immunizing rabbits with the coupled peptide DAASKPS⁸⁵⁹GEGSFSK.

Li X., Zhong L., Wang Z., Chen H., Liao D., Zhang R., Zhang H, & Kang T. (2018). Phosphorylation of IRS4 by CK1γ2 promotes its degradation by CHIP through the ubiquitin/lysosome pathway. Theranostics, 8(13), 3643-3653.

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Regulation of GPER/PI3K/Akt Signaling

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Rb1 (purity >98%) was purchased from ChromaDex Corp. (USA). Antibodies against GPER (ab188790) were purchased from Abcam (USA). Antibodies against PI3K (bs-2067R) and phosphorylated PI3K (p-PI3K) (bs-10657R) were purchased from Bioss (Beijing, China). Antibodies against Akt (sc-8312), p-Akt (sc-135651), and β-actin (sc-47778) were purchased from Santa Cruz Biotechnology, Inc. (CA, USA). Antibodies against Bax (BA0315-2) and Bcl-2 (BA0412) were purchased from Boster (Wuhan, China). Enzyme-linked immunosorbent assay (ELISA) kits for detection of IL-6 (H007), IL-1β (H002), and TNF-α (H052) were obtained from Nanjing Jiancheng Bioengineering Institute. Secondary goat anti-rabbit (A0208), goat anti-mouse IgG-HRP (A0216), and cy3-labled goat anti-mouse IgG-HRP (A0521) antibodies were purchased from Beyotime (Shanghai, China). Total Protein Extraction Kits (P0027) were purchased from Beyotime (Shanghai, China). PI/Annexin V-FITC Apoptosis Detection Kits were purchased from JingMei Biotech (Beijing, China). Antagonist (G-15) (3678) for GPER and agonist (740 Y-P) for PI3K (1983) were obtained from R&D system (USA).

Yang G., Zhuo J., Lin Y., Zhang M., Liu L., Chen X, & Gao R. (2019). Ginsenoside Rb1 Prevents Dysfunction of Endothelial Cells by Suppressing Inflammatory Response and Apoptosis in the High-Fat Diet Plus Balloon Catheter-Injured Rabbit Model via the G Protein-Coupled Estrogen Receptor-Mediated Phosphatidylinositol 3-Kinases (PI3K)/Akt Pathway. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 7407-7417.

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Molecular Regulation of Cell Cycle Proteins

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Psoralen (purity > 98%, HPLC, Supplementary Figure 1) was purchased from Nanjing Jing Zhu Bio-Technology Co., Ltd. (Nanjing, China). Primary antibody to cyclin D1(2978), cyclin E1(20808), CDK1(77055), p27(3688), phospho-AMPKα(2535), AMPKα(2532), phospho-AKT(4060), phospho-mTOR(2971), mTOR(2983), phospho-4E-BP1(2855), 4E-BP1(9644), phospho-p70S6 kinase(9204), p70S6 kinase(2708) were purchased from Cell Signaling Technology (Danvers, MA, United States). Antibody to cyclin A2 (ab181591) was purchased from Abcam (Cambridge, MA, United States), AKT (sc-8312) and β-actin (sc-69879) were purchased from Santa Cruz Biotechnology (Dallas, TX, United States).

Zhou W., Chen X., Zhao G., Xu D., Jiang Z., Zhang L, & Wang T. (2018). Psoralen Induced Liver Injury by Attenuating Liver Regenerative Capability. Frontiers in Pharmacology, 9, 1179.

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Western Blot Analysis of Adipocyte Markers

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Cells were scraped with RIPA (Radio Immunoprecipitation Assay) buffer (Beyotime, Shanghai, China) and lysates were subjected to SDS-PAGE and transferred to the PVDF (Polyvinylidene fluoride, Millipore, Burlington, MA, USA). Polyacrylamide gels were used to separate and mark proteins of different sizes. The proteins were then transferred to a PVDF membrane. Next, the membrane was soaked in 5% skim milk for 2 h and then incubated with primary antibodies overnight at 4 °C. After that, membranes were washed in Tris-buffered saline with Tween 20 and subsequently incubated with horseradish peroxidase-conjugated secondary antibodies. Finally, the stripes of target proteins were visualized by the enhanced chemiluminescent substrate (Millipore, MA) and observed using Gel Doc XR System (Bio-Rad). The densities of the brands were analyzed using Image Lab software (Bio-Rad). Target proteins were probed with primary antibodies (anti-AQP3, ab125219, Abcam, 1:1000; PPARγ, #2435, CST, 1:1000; FASN, sc-20140, Santa Cruz, 1:200; FABP4, sc-18661, Santa Cruz, 1:200; Akt, sc-8312, Santa Cruz, 1:200; p-Akt, sc-7985-R, Santa Cruz, 1:200; cyclin D, sc-753, Santa Cruz, 1:500; cyclin E, sc-247, Santa Cruz, 1:500; β-actin, sc-130656 Santa Cruz, 1:1000).

Wang X., Yang J., Yao Y., Shi X., Yang G, & Li X. (2020). AQP3 Facilitates Proliferation and Adipogenic Differentiation of Porcine Intramuscular Adipocytes. Genes, 11(4), 453.

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Protein Expression Assay Protocol

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H₂O₂ (30%) was obtained from Sigma (St Louis, MO, United States). Antibodies targeting α-SMA (ab5694), fibronectin (ab2413) and Col1A (ab96723) were purchased from Abcam (Cambridge, United Kingdom). Akt (sc-8312) and PARP/c-PARP (sc-7150) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). GAPDH (#2118), phospho-Akt (#4060), and caspase-3/c-caspase-3 (#9665T) antibodies, horseradish peroxidase (HRP)-linked anti-rabbit IgG (#7074) and HRP-linked anti-mouse IgG (#7076) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Yang B.Y., Zhou Z.Y., Liu S.Y., Shi M.J., Liu X.J., Cheng T.M., Deng G.Y., Tian Y., Song J, & Li X.H. (2022). Porous Se@SiO2 Nanoparticles Enhance Wound Healing by ROS-PI3K/Akt Pathway in Dermal Fibroblasts and Reduce Scar Formation. Frontiers in Bioengineering and Biotechnology, 10, 852482.

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Uterus Protein Expression Analysis

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Proteins were extracted from the whole uterus from 20-week-old female mice and western blot was performed to measure PTEN, AKT, P-AKT, and p27 levels as previously described (Vignarajan et al. 2014) . The antibodies used were PTEN (138G6, Cell Signaling, 1:500), AKT (sc-8312, Santa Cruz Biotechnology, 1:500), P-AKT (sc-7985, Santa Cruz Biotechnology, 1:300), p27 (sc-528, Santa Cruz Biotechnology, 1:500), and B-actin (ab8229, Abcam, Cambridge, UK, 1:3000).

Choi J.P., Desai R., Zheng Y., Yao M., Dong Q., Watson G., Handelsman D.J, & Simanainen U. (2015). Androgen actions via androgen receptor promote PTEN inactivation induced uterine cancer. Endocrine-related cancer, 22(5).

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