Briefly, cells were harvested and lysed in 1 × loading buffer followed by separation on a 12% polyacrylamide gel. Proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane. The membranes were blocked and then incubated with primary antibodies, followed by a corresponding AP-conjugated secondary antibody or HRP-conjugated secondary antibody. Proteins that used AP-conjugated secondary antibody were visualized using the substrates nitroblue tetrazolium (NBT) (#N6876; Sigma) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP; #11585002001; Roche). Proteins that used HRP-conjugated secondary antibody were incubated by hypersensitive ECL chemiluminescence detection kit (#B500022; Proteintech) and visualized using the Azure C500 Infrared Imaging System (Azure Biosystems, Dublin, CA, United States).
Azure c500 infrared imaging system
Manufactured by Azure Biosystems
Sourced in Ireland, United States
The Azure C500 Infrared Imaging System is a laboratory instrument designed for infrared imaging applications. It utilizes infrared technology to capture and analyze images. The core function of the system is to provide high-quality infrared imaging capabilities for research and analysis purposes.
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Lab products found in correlation
2 protocols using azure c500 infrared imaging system
1
Western Blot Analysis Protocol
2
Western Blot Protein Analysis Protocol
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The cells were harvested and boiled in 1 × loading buffer (0.08 mol/L Tris, pH 6.8, with 2.0% SDS, 10% glycerol, 0.1 mol/L dithiothreitol, and 0.2% bromophenol blue) followed by separation by SDS-PAGE electrophoresis. Proteins were transferred to PVDF membranes and blocked with 5% skim milk powder at room temperature for 1 h. Next, the PVDF membranes were washed with TBST and incubated with primary antibodies against target proteins at 4 °C overnight, followed by two washes with TBST. PVDF membranes were incubated with the appropriate secondary antibodies at room temperature for 1 h and washed three times with TBST. The blots were reacted with nitroblue tetrazolium (NBT) and 5-bromo-4-chloro-3′-indolylphosphate (BCIP), or were visualized by using the Azure C500 Infrared Imaging System (Azure Biosystems, Dublin, CA, USA).
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