Total proteins (20 μg) extracted from thoracic aortas were resolved on 10% SDS-polyacrylamide gels and electrotransferred onto nitrocellulose membranes by electrophoresis. The membranes were blocked in 5% nonfat milk and TBS containing 0.3% Tween-20, subsequently incubated overnight with polyclonal rabbit anti-rat chemerin (1 : 1000, Abcam, UK), NF-κBp65 (1 : 1000, Cell Signaling Technology, USA), PCNA (1 : 500, Santa Cruz, USA), p-p38-MAPK (1 : 1000, Santa Cruz, USA), p-ERK 1/2 (1 : 1000, Santa Cruz, USA), p-JNK (1 : 500, Cell Signaling Technology, USA), and β-actin (1 : 1000, Santa Cruz, USA). The blots were incubated in 1 : 6000 dilution of goat anti-rabbit horseradish peroxidase-conjugated secondary antibodies. 100 μl ECL solution was added in each sample to detect the protein signal with the Quant RT ECL cold CCD imaging system (General Electric, USA).
Quant rt ecl cold ccd imaging system
Manufactured by GE Healthcare
Sourced in United States
The Quant RT ECL cold CCD imaging system is a laboratory equipment designed for the detection and quantification of chemiluminescent signals. It utilizes a charge-coupled device (CCD) camera to capture images of chemiluminescent samples, allowing for sensitive and accurate detection of target analytes.
Automatically generated - may contain errors
Lab products found in correlation
Nf κb p65, by Cell Signaling Technology (1 mentions)
Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions)
Polyclonal rabbit anti rat chemerin, by Abcam (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
P p38 mapk, by Santa Cruz Biotechnology (1 mentions)
P erk1 2, by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Abcam (1 mentions)
Bcl2 associated x (bax), by Abcam (1 mentions)
P p38 mapk, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
2 protocols using quant rt ecl cold ccd imaging system
1
Protein Expression Analysis in Aortic Tissues
2
HL-1 Cardiomyocyte Western Blot Analysis
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For Western blot analysis, 20 μg of total protein extracted from the HL-1 cardiomyocytes after the cells were treated for 24 h was resolved on 10% SDS-polyacrylamide gels and electrotransferred onto nitrocellulose membranes. The membranes were blocked with 5% nonfat milk in TBS containing 0.3% Tween-20 and then incubated overnight with polyclonal rabbit anti-mouse antibodies against Bcl-2 (1:1000 dilution, Abcam, USA), Bax (1:500 dilution, Abcam), caspase-3 (1:1000 dilution, Cell Signaling, USA), and p-p38 MAPK (1:1000 dilution, Cell Signaling). The polyclonal rabbit anti-mouse GAPDH antibody (1:1000 dilution, Abcam) served as control. The goat anti-rabbit horseradish peroxidase-conjugated secondary antibody was subsequently added. ECL was administered to detect protein signals using Quant RT ECL cold CCD imaging system (General Electric, USA).
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