Amlodipine and gefitinib were purchased from Selleck Chemicals (Houston, TX, United States) and TopScience (Shanghai, China), respectively. Propidium iodide (PI) was obtained from Sigma-Aldrich (St. Louis, MO, United States). The Annexin V/FITC-PI apoptosis detection kit was obtained from BD Biosciences (San Jose, CA, United States of America). RPMI 1640 and FBS were purchased from Biological Industries (Beit Haemek, Israel). The enhanced chemiluminescence (ECL) reagent was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Antibodies against phospho-EGFR, phospho-PDK1 (Ser241), PDK1, phospho-Akt (Ser473), phospho-Akt (Thr308), Akt, phospho-mammalian target of rapamycin complex 1 (mTORC1) (Ser2448), mTOR, phospho-glycogen synthase kinase 3 (GSK-3) β, phospho-p70 S6K (Thr389), p27, cyclin D1, phospho-retinoblastoma protein (Rb), phospho-extracellular signal-regulated kinase (ERK) 1/2, phospho-c-Raf, β-actin, and horseradish peroxidase (HRP)-conjugated goat anti-rabbit and horse anti-mouse secondary antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, United States). Antibody specific for p21 was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, United States).
Horse anti mouse secondary antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Horse anti-mouse secondary antibodies are designed for the detection and quantification of mouse primary antibodies in various immunoassays, such as Western blotting, ELISA, and immunohistochemistry. These antibodies are generated in horses and specifically recognize and bind to mouse immunoglobulins, allowing for the amplification and visualization of mouse primary antibody signals.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Sartorius (3 mentions)
Rpmi 1640, by Sartorius (3 mentions)
β actin, by Cell Signaling Technology (3 mentions)
Erk1 2, by Cell Signaling Technology (2 mentions)
Enhanced chemiluminescence ecl reagent, by Thermo Fisher Scientific (2 mentions)
Gefitinib, by Selleck Chemicals (2 mentions)
Goat anti rabbit secondary antibody, by Cell Signaling Technology (2 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Phospho c raf, by Cell Signaling Technology (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Amlodipine, by Selleck Chemicals (1 mentions)
Phospho retinoblastoma protein rb, by Cell Signaling Technology (1 mentions)
Antibody specific for p21, by Santa Cruz Biotechnology (1 mentions)
Phospho p70s6k, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Pi3k p110β, by Cell Signaling Technology (1 mentions)
Poly adp ribose polymerase parp, by Cell Signaling Technology (1 mentions)
P p70 s6 kinase, by Cell Signaling Technology (1 mentions)
5 protocols using horse anti mouse secondary antibody
1
Amlodipine and Gefitinib Apoptosis Assay
2
Investigating Apoptosis and Cell Signaling Pathways
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Lansoprazole and gefitinib were purchased from Selleck Chemicals (Houston, TX, United States) and Target Molecule Corp. (Boston, MA, United States), respectively. Monodansylcadaverine (MDC) and propidium iodide (PI) were obtained from Sigma-Aldrich (St. Louis, MO, United States). A FITC-Annexin V apoptosis detection kit was obtained from BD Bioscience (San Josè, CA, United States). RPMI 1640 and FBS were purchased from the Biological Industries (Beit Haemek, Israel). Enhanced chemiluminescence (ECL) reagent was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Antibodies specific for p27, p-Rb, cyclin D1, LC3B, p-signal transducer and activator of transcription (Stat) 3, phosphatidylinositol 3-kinase (PI3K)-p110α, PI3K-p110β, p-mammalian target of rapamycin complex 1 (mTORC1) (Ser2448), Akt, p-Akt (Ser473), p-p70 S6 kinase (S6K) (Thr389), p-glycogen synthase kinase 3 (GSK-3)β, poly (ADP-ribose) polymerase (PARP), caspase-3, K-Ras, p-extracellular signal-regulated kinase (ERK)1/2, p-c-Raf, Ki67, β-actin, and horseradish peroxidase-conjugated goat anti-rabbit and horse anti-mouse secondary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, United States). Antibodies specific for Bcl-2 and Bax were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, United States).
3
Molecular Mechanisms of Anticancer Agents
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PKM2-IN-1 and NCT-503 were purchased from MedChemExpress (Princeton, NJ, United States). RPMI 1640 and FBS were purchased from Biological Industries (Beit Haemek, Israel). Propidium iodide (PI) was obtained from Sigma-Aldrich (St. Louis, MO, United States). The enhanced chemiluminescence (ECL) reagent was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Antibodies against cyclin B1, cdc2, caspase 3, poly (ADP-ribose) polymerase (PARP), Bax, B-cell lymphoma 2 protein (Bcl-2), pyruvate kinase (PK) M2, phospho-AMP-activated protein kinase (AMPK)α (Thr172), phospho-mTOR (Ser2448), phospho-p70 ribosomal protein S6 Kinase (p70S6K) (Thr389), phospho-histone H2AX (Ser139) (γ-H2AX), p-ATM (Ser 1981), β-actin, and HRP-conjugated goat anti-rabbit and horse anti-mouse secondary antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, United States). The anti-p-Chk2 (Thr68) antibody was purchased from Abcam (Cambridge, UK). An antibody specific for PHGDH was purchased from Absin Bioscience Inc. (Shanghai, China). Antibodies against p-p53 (ser15), p53, p21 and Ki67 were purchased from Proteintech Group, Inc (Rosemont, IL, United States). Antibodies against glucose transporter type 1 (GLUT1) was purchased from Abclonal (Woburn, MA, United States).
4
Protein Expression Analysis by Western Blot
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A protein extraction kit (KeyGEN, Nanjing, China) was used to lyse the cells. Protein concentrations were measured with a BCA kit (Beyotime, Shanghai, China). The proteins were separated by SDS–PAGE and transferred to a PVDF membrane. The membrane was blocked with 5% skim milk for 1 h at room temperature and was then incubated with antibodies against AMPK, p-AMPK (Thr172), mTOR, p-mTOR (Ser2448), NF-κBp65, phospho-NF-κBp65 (Ser536), p38 MAPK, phospho-p38 MAPK (Thr180/Tyr182), p70S6 kinase, phospho-p70 S6 kinase (Thr389), AKT, phospho-AKT (Ser473), Smad2, phospho-Smad2 (Ser465/467), β-actin, Beclin-1, LC3B, SQSTM1/p62 (Cell Signaling Technology, Beverly, MA, USA), fibronectin, α-smooth muscle actin (α-SMA), GAPDH, and human collagen (type I) (Thermo Fisher Scientific, Rockford, IL, USA) at 4 °C overnight. The membrane was then incubated with a goat anti-rabbit secondary antibody or a horse anti-mouse secondary antibody (Cell Signaling Technology, Boston, MA, USA) at room temperature for 1 h. Finally, the signals were examined using a chemiluminescence kit (Bio–Rad, Hercules, CA, USA).
5
Western Blot Analysis of Cellular Proteins
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