Intracellular Ca2+ concentration was determined according to the Fluo-4, AM kit protocol. Briefly, K562 cells (1 × 106 cells/mL) were inoculated into 24-well plates and maintained for 24 h. Then, the cells were treated with AMP-17 (40, 60, and 80 μg/mL) or Ara-C (3.5 mg/mL) for 24 h. After incubation, the cells were collected and washed twice with PBS. Subsequently, 1-[2-Amino-5-(2,7-difluoro-6-hydroxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5-methylphenoxy)ethane-N,N,N′,N′-tetraacetic acid, pentaacetox-ymethyl ester (Fluo-4/AM, Solarbio, China) working solution was added to each group of cells and incubated in the dark for 20 min at 37 °C. After that, 5 times the volume of HBSS (Solarbio, China) buffer containing 1% FBS was added to each group of cells and incubated for 40 min. Finally, the cells were washed three times with HEPES buffer saline (Solarbio, China) and resuspended, incubated at 37 °C for 10 min, and then the Ca2+ concentration was measured by flow cytometry (Beckman, USA).
Hepes buffer saline
Manufactured by Solarbio
Sourced in China, United States
HEPES buffer saline is a buffer solution that maintains a stable pH environment. It is commonly used in cell culture and biochemical applications to maintain physiological pH levels.
Automatically generated - may contain errors
Lab products found in correlation
Hanks balanced salt solution (hbss), by Solarbio (2 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Fluo 4 am, by Solarbio (1 mentions)
Opti mem 1 reduced serum medium, by Thermo Fisher Scientific (1 mentions)
Mitosox red mitochondrial superoxide indicator m36008, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Anti gapdh d4c6r, by Cell Signaling Technology (1 mentions)
2 protocols using hepes buffer saline
1
Intracellular Calcium Determination in K562 Cells
2
Naringenin and Homocysteine Modulate Endothelial Function
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Naringenin (B21596, for in vitro experiments, purity > 98%, ASB-00014206–001, for in vivo experiments, purity > 98%) and homocysteine (S20422, purity > 90%) were purchased from Shanghai Source Leaf Biological Technology Co., Ltd. Endothelial cell medium (SC-1001) was sourced from ScienCell, Fluo-3 AM (F8841). HEPES buffer saline and Hank’s balanced salt solution (H1025) were purchased from Beijing Solarbio Science & Technology Co., Ltd. MitoSOX™ red mitochondrial superoxide indicator (M36008), TRIzolTM reagent (15596026), trypsin (15050065), Lipofectamine™ RNAiMAX Transfection Reagent (13778100), Opti-MEM™ I Reduced Serum Medium (31985070), siRNA included PRKAA1 (s101), SIRT1 (s223592) and Silencer™ Select Negative Control No. 1 siRNA (4390843) were purchased from Thermo Fisher Scientific (MA, USA). Antibodies; anti-AMPKα (D5A2), anti-Sirt1 (C14H4), anti-NMDA-R1 (D65B7) and anti-GAPDH (D4C6R) were sourced from Cell Signaling Technology, whereas anti-eNOS (ab76198) was purchased from Abcam.
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