Ab146428

Manufactured by Abcam

Ab146428 is a laboratory product offered by Abcam. It is a research-use tool designed for scientific applications. The core function of this product is to facilitate laboratory experimentation and analysis, but a detailed description cannot be provided while maintaining an unbiased and purely factual approach.

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Lab products found in correlation

Endothelial cell growth medium 2 bulletkit, by Lonza (1 mentions) C2c12 cell, by American Type Culture Collection (1 mentions) Huvecs, by Lonza (1 mentions) Humulin r, by Eli Lilly (1 mentions) Pglv3 h1 gfp puro vector, by GenePharma (1 mentions) Sodium orthovanadate, by Fujifilm (1 mentions) Ab182651, by Abcam (1 mentions) β actin 13e5, by Cell Signaling Technology (1 mentions) Atg16l1, by Cell Signaling Technology (1 mentions)

3 protocols using ab146428

Endothelial and Muscle Cell Response to Norepinephrine

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Human umbilical vein endothelial cells (HUVECs, Lonza) were cultured with Endothelial Cell Growth Medium-2 BulletKit (Lonza), and C2C12 cells (ATCC, CRL1772) were cultured with Dulbecco’s Modified Eagle Medium (DMEM) and differentiated according to the manufacturer’s instruction. Arginine (5 mM for a total of 73 h; Merck A5006), SC79 (AKT activator; 10 μM for a total of 73 h; abcam, ab146428), 3-hydroxybutyrate (3-HB; 10 mM for a total of 73 h; Sigma-Aldritch, 54,920), or DETA NONOate (a NO donor; 100 μM for 73 h; abcam, ab144627) was administered 1 h before norepinephrine (NE) was introduced (10 ng/ml for a total of 72 h; Daiichi Sankyo), and the medium containing these reagents was refreshed every 24 h. For the Western blot study for phosphor AKT detection, HUVECs were serum starved with 0.5% FBS for a total of 4 h; at the start of the starvation, 3-HB (10 mM Sigma-Aldritch, 54920) was added in some plates. One hour after the starvation, NE (10 ng/ml) was added for a total of 3 h. Insulin (HumulinR, Lilly, 100 nmol/l) was administered 10 min before collecting cells.

Nakao M., Shimizu I., Katsuumi G., Yoshida Y., Suda M., Hayashi Y., Ikegami R., Hsiao Y.T., Okuda S., Soga T, & Minamino T. (2021). Empagliflozin maintains capillarization and improves cardiac function in a murine model of left ventricular pressure overload. Scientific Reports, 11, 18384.

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Investigating EphB3 in Esophageal Squamous Cell Carcinoma

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HEEC cells (BNCC337729) were supplied by the Beina Chuanglian Biotechnology Institute (Beijing, China). The ESCC cell line TE-1 (TCHu89) was sourced from the Cell Bank of the Chinese Academy of Sciences and TE10, TE-13, KYSE-150, KYSE-450 were gifts from Professor Sun of Nanjing Medical University. Dulbecco’s modified Eagle’s medium (containing 10% fetal bovine serum and 1% penicillin and streptomycin) was used to culture the cells in reagents and supplements supplied by GIBCO Invitrogen Inc. (Carlsbad, CA, USA). The ESCC cells were treated with 5 µg/mL AKT activator SC79 (ab 146428, Abcam) for 48 h. To generate cell lines that stably suppressing EphB3, EphB3-targeting shRNA or scrambled shRNA sequences were cloned into the linear lentiviral vector pgLV3/H1/GFP + Puro Vector provided by GenePharma (Shanghai); cells were then infected with the recombinant lentivirus. Next, they were selected after the addition of puromycin (2 µg/mL) to the culture medium for 14 days. shRNAs sequences were designed using software provided by Invitrogen and details are listed in Table S1.

Tang L., Wang L., Yu T., Xu T., Zhang L, & Shu Y. (2022). Knockdown of EphB3 inhibits cell proliferation partly through the AKT signaling pathway and represses epithelial-mesenchymal transition in esophageal squamous cell carcinoma. Translational Cancer Research, 11(1), 85-98.

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Antibody-Based Signaling Pathway Analysis

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The RCS are listed in Table S1.
Antibodies against Akt (#9272S), phospho-Akt (Ser473) (D9E) (#4060), phospho-Akt (Thr308) (D25E6) (#13038), EGFR (D38B1) (#4267), phospho-EGFR (Tyr1068) (D7A5) (#3777), IR (4B8) (#3025), phospho-IR (Tyr1150/1151) (#3021), Bad (D24A9) (#9239), phospho-Bad (Ser136) (D25H8) (#4366), phospho-GSK3β (Ser9) (D85E12) (#5558), β-actin (13E5) (#5125), phospho-ATG16L1 (Ser278) (#84300), and ATG16L1 (D6D5) (#8089) were purchased from Cell Signaling Technology. Antibodies against PI3K (610045) and GSK3β (610201) were purchased from BD Transduction Laboratories. Antibodies against phospho-PI3K p85a (Tyr607) (ab182651) and SC-79 (ab146428) were purchased from Abcam. Antibodies against FLAG (DYKDDDDK) (A8592) were purchased from Sigma–Aldrich. Antibodies against LC3 (PM036) were purchased from Medical & Biological Laboratories. The following antibodies were purchased from GE Healthcare: anti-mouse IgG-horseradish peroxidase (NA9310) and anti-rabbit IgG-horseradish peroxidase (NA9340). Recombinant human EGF (236-EG) was purchased from R&D Systems. Recombinant human insulin (096-03443), sodium orthovanadate, and wort were purchased from Fujifilm–Wako. Recombinant human PI3K (p110a/p85a) (V1721) was purchased from Promega. BYL719 (S2814) was purchased from Selleck.

Morimoto A., Takasugi N., Pan Y., Kubota S., Dohmae N., Abiko Y., Uchida K., Kumagai Y, & Uehara T. (2024). Methyl vinyl ketone and its analogs covalently modify PI3K and alter physiological functions by inhibiting PI3K signaling. The Journal of Biological Chemistry, 300(3), 105679.

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