The concentration of protein in the sample was measured using a BCA Protein Quantification Kit (Solarbio, Beijing, China). Forty micrograms samples were mixed with 10% SDS-PAGE before being transferred to PVDF membranes (Millipore, Massachusetts, USA). The membranes were blocked with 5% degreased milk powder for 1 h. The primary antibody of each protein was diluted with 5% BSA as follows: rabbit anti-rat Bax(1:800, orb224426, Biorbyt, Cambridge, UK), Bcl-2(1:800, orb228150, Biorbyt), caspase-3(1:800, orb10231, Biorbyt), PIK3CA (1:800, orb228203, Biorbyt), PI3K (1:800, orb137259, Biorbyt), p-PI3K (1:800, orb338965, Biorbyt), Akt (1:800, orb213545, Biorbyt), p-Akt (1:800, orb222951, Biorbyt), and β-actin (1:2000, orb178392, Biorbyt). All primary antibodies were reacted overnight at 4 °C, then incubated with the secondary goat anti-rabbit Ig G (1:1500, ab6721, Abcam, Beijing, China) for 1 h.
Orb213545
Manufactured by Biorbyt
Sourced in United Kingdom
Orb213545 is a laboratory equipment product from Biorbyt. It is designed for a core function, but no further details on its intended use can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Ab6721, by Abcam (2 mentions)
Orb178392, by Biorbyt (2 mentions)
Orb224426, by Biorbyt (2 mentions)
Orb228150, by Biorbyt (2 mentions)
Orb10231, by Biorbyt (2 mentions)
Orb228203, by Biorbyt (2 mentions)
Orb137259, by Biorbyt (2 mentions)
Orb338965, by Biorbyt (2 mentions)
Orb222951, by Biorbyt (2 mentions)
Bca protein quantification kit, by Solarbio (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Bca protein quanti cation kit, by Solarbio (1 mentions)
2 protocols using orb213545
1
Protein Quantification and Western Blot
2
Protein Expression Analysis of HK-2 Cells
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The concentration of protein sample was measured by a BCA protein quanti cation kit (Solarbio, Beijing, China). 40 μg samples were mixed with 10% SDS-PAGE electrophoresis before transferred to PVDF membrane (Millipore, Massachusetts, USA). The membranes were blocked with 5% degreased milk powder for 1 h. The primary antibody of each protein was diluted with 5% BSA. And the rabbit anti-rat Bax 1:800, orb224426, Biorbyt, Cambridge, UK Bcl-2 1:800, orb228150, Biorbyt caspase-3 1:800, orb10231, Biorbyt PIK3CA (1:800, orb228203, Biorbyt), PI3K (1:800, orb137259, Biorbyt), p-PI3K (1:800, orb338965, Biorbyt), AKT (1:800, orb213545, Biorbyt), p-AKT (1:800, orb222951, Biorbyt), β-actin (1:2000, orb178392, Biorbyt). All primary antibodies were reacted overnight at 4°C, then incubated with the second antibody goat anti-rabbit Ig G (1:1500, ab6721, Abcam, UK) for 1h.
Cell Culture HK-2cells (Shanghai Institute of Cell Research), human renal tubular epithelial cells, were cultured in DMEM/F12 with 10% fetal bovine serum and 1% penicillin-streptomycin (GIBCO, Invitrogen, Beijing, China) at 37°C, 5% CO 2 .
Cell Culture HK-2cells (Shanghai Institute of Cell Research), human renal tubular epithelial cells, were cultured in DMEM/F12 with 10% fetal bovine serum and 1% penicillin-streptomycin (GIBCO, Invitrogen, Beijing, China) at 37°C, 5% CO 2 .
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