Prostate cancer tissue was used as a positive control in western blot analysis of purified CTRP3 protein. The specimen was collected from a prostate cancer patient confirmed by pathology. This study was ratified by the ethics committees of Central Hospital of Longgang District. The piece of tissue was crushed in liquid nitrogen, and then used to extract protein. Total cellular proteins were prepared from treated or untreated cells by lysing cells in lysis buffer (CelLytic, Sigma-Aldrich). Lysates were resolved on 10% or 12.5% polyacrylamide gels. Proteins were transferred to a PVDF membrane (Bio-Rad, Hercules, CA, USA), which was blocked with 5% milk containing TBS-T buffer (0.05% Tween-20) and incubated with primary antibodies at 4°C overnight. After washes with TBST, the membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit IgG, donkey anti-goat IgG, or goat anti-mouse IgG antibodies (Jackson, USA). Immunoreactive bands were visualized with the Western Breeze chromogenic detection system (Invitrogen).
Western breeze chromogenic detection system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Western Breeze chromogenic detection system is a comprehensive solution designed for western blot analysis. The system provides all the necessary components for the detection and visualization of protein targets, including reagents, membranes, and accessories. The core function of the Western Breeze system is to enable the detection of proteins separated by gel electrophoresis and transferred to a membrane, allowing for the identification and quantification of specific protein targets.
Automatically generated - may contain errors
Lab products found in correlation
Cellytic, by Merck Group (2 mentions)
Donkey anti goat igg, by Jackson ImmunoResearch (1 mentions)
Goat anti mouse igg antibody, by Jackson ImmunoResearch (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Proteinase inhibitor cocktail, by Merck Group (1 mentions)
Ab36870, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Enhanced chemi luminescence (ecl), by Bio-Rad (1 mentions)
2 protocols using western breeze chromogenic detection system
1
Western Blot Analysis of CTRP3 in Prostate Cancer
2
Western blot analysis of C1QTNF3 protein
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Cancer or paired normal tissue (0.2 g) was crushed in liquid nitrogen and lysed in RIPA lysis buffer (CelLytic, Sigma-Aldrich) in the presence of a proteinase inhibitor cocktail (Merck Millipore, USA). Total protein extracts were separated by SDS-PAGE and transferred to PVDF membranes (Merck Millipore, USA). Immunoblotting was done with rabbit polyclonal antibody against C1QTNF3 (ab36870, Abcam, 1:1500 dilution) in accordance with the manufacturer's instruction. Signals were visualized using enhanced chemiluminescent substrate (ECL, BioRad, Richmond, CA, USA) and the Western Breeze chromogenic detection system (Invitrogen).
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