Total proteins were extracted from the aortic segments using a lysis buffer (Beyotime, China). These lysates were then separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE, Invitrogen, USA), transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA), and then incubated with primary antibodies against chemerin (1 : 1000, ab103153, Abcam, USA), CMKLR1 (1 : 1000, AP06779, Origene, USA), MMP-2 (1 : 1000, ab86607, Abcam, USA), MMP-9 (1 : 1000, ab38898, Abcam, USA), and GAPDH (1 : 10000, ab8245, Abcam, USA) overnight at 4°C. The next day, the blots were incubated with horseradish peroxidase- (HRP-) conjugated AffiniPure goat anti-rabbit IgG (1 : 5000, SA00001-2, Proteintech, China) or anti-mouse IgG (1 : 5000, SA00001-1, Proteintech, China) for 1h at room temperature. Proteins were visualized using Amersham Imager (Cytiva, USA) and quantified using ImageJ software.
Amersham imager
Manufactured by Cytiva
Sourced in United States
The Amersham Imager is a versatile lab equipment designed for a range of imaging applications. It enables the capture and analysis of various types of blots, gels, and other samples using different imaging techniques, including chemiluminescence, fluorescence, and colorimetric detection. The Amersham Imager provides high-quality imaging results and supports a variety of sample types and applications within the laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Ab86607, by Abcam (1 mentions)
Ab103153, by Abcam (1 mentions)
Ap06779, by OriGene (1 mentions)
Sds page, by Thermo Fisher Scientific (1 mentions)
Ab8245, by Abcam (1 mentions)
Sa00001 1, by Proteintech (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
Ab38898, by Abcam (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Ab62344, by Abcam (1 mentions)
E ab 1003, by Elabscience (1 mentions)
Ab18256, by Abcam (1 mentions)
E ab 20058, by Elabscience (1 mentions)
Nbp3 03885, by Novus Biologicals (1 mentions)
Wbkls0100, by Merck Group (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
Ab13556, by Abcam (1 mentions)
2 protocols using amersham imager
1
Aortic Protein Expression Analysis
2
Investigating Cell Death Signaling in Aortic Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein lysates from aortic tissues were extracted and protein concentrations were tested by BCA Protein Assay Kit (Solarbio, China). Proteins (20 µg per lane) were separated with SDS-PAGE and were transferred to PVDF membranes (Millipore, USA). Membranes were blocked for 1 hour in 5% skimmed milk and incubated overnight at 4°C with the antibodies as follows: TLR4 (1:500, ab13556, Abcam, USA), HMGB1 (1:1000, ab18256, Abcam, USA), RIP1 (1:1000, 3493, Cell Signaling, USA), RIP3 (1:1000, ab62344, Abcam, USA), MLKL (1:1000, NBP3-03885, Novus, USA), and β-actin (1:2000, E-AB-20058, Elabscience, China). Next, the membranes were rinsed in TBST and incubated with secondary antibody (horseradish peroxidase-(HRP-) conjugated goat anti-rabbit IgG, 1:5000, E-AB-1003, Elabscience, China) for 1 hour at room temperature. Signals were tested by enhanced chemiluminescence (WBKLS0100, Millipore, Germany) and exposed with Amersham Imager (Cytiva, USA). Protein strips were measured with ImageJ software.
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