At first, the cell or tissue protein were extracted using the RIPA lysis buffer (P0013B, Beyotime). The protein concentration was determined by the BCA protein assay kit. All samples were prepared at a final concentration of 1 μg/μl in loading buffer (CW0027S, CwBio). Protein samples (20 μg) were loaded into a 10–15% SDS-polyacrylamide gel in the Bio-Rad Electrophoresis System to separate the proteins with different molecular weight. Then, the proteins in the gel were transferred to polyvinylidene difluoride (PVDF) membranes. After blocking in 5% bovine serum albumin (BSA) solution, the membranes were incubated with primary antibodies (Integrin-αL, ab186873, Abcam; Integrin-β2, ab131044, Abcam; C1q, ab71940, Abcam; Integrin-β3, ab75872, Abcam; CD11b, ab133357, Abcam; CD45, ab10558, Abcam; cleaved caspase-3, #9661s, Cell signaling technology; CD44, ab189524, Abcam; HO-1, ab68477, Abcam; β-tubulin, CW0098, CwBio; iNOS, ab15323, Abcam; CD206, ab125028, Abcam; Arginase-1, 93668s, Cell signaling technology; Histone H3, #9715, Cell signaling technology; GM130, 610,822, BD Biosciences) at 4 °C overnight. After incubation with the corresponding secondary antibodies, the PVDF membranes were imaged using Western chemiluminescent horseradish peroxidase (HRP) substrate (Millipore) with an imaging system (Tanon 4600, Shanghai).
Ab186873
Manufactured by Abcam
Ab186873 is an antibody that recognizes the vascular endothelial growth factor receptor 2 (VEGFR2) protein. VEGFR2 is a receptor tyrosine kinase that plays a key role in the regulation of angiogenesis, the process of new blood vessel formation. This antibody can be used in various research applications for the detection and study of VEGFR2.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ab133357, by Abcam (1 mentions)
Ab71940, by Abcam (1 mentions)
Ab75872, by Abcam (1 mentions)
Ab131044, by Abcam (1 mentions)
Ab125028, by Abcam (1 mentions)
Ab189524, by Abcam (1 mentions)
Ab10558, by Abcam (1 mentions)
Ab68477, by Abcam (1 mentions)
Cw0027s, by CWBIO (1 mentions)
Cw0098, by CWBIO (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Ab15323, by Abcam (1 mentions)
Electrophoresis system, by Bio-Rad (1 mentions)
Ab64693, by Abcam (1 mentions)
Ab8226, by Abcam (1 mentions)
Ab6640, by Abcam (1 mentions)
Anti p nf κb p65, by Cell Signaling Technology (1 mentions)
Anti actinin 4, by Cell Signaling Technology (1 mentions)
Anti integrin β1, by Cell Signaling Technology (1 mentions)
2 protocols using ab186873
1
Comprehensive Protein Expression Analysis
2
Western Blot Analysis of Immune and Adhesion Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The protein lysates from the cells and kidney tissues were prepared following standard protocols, and the protein content was determined using the BCA protein assay kit (ThermoFisher). And then the proteins samples were separated by Bis-Tris Gel (Invitrogen) and transferred onto PVDF membranes (Millipore) using a wet- transfer system. Membranes were blocked in 5% BSA in TBS-T for 1 h at room temperature and were incubated with primary antibodies overnight at 4 °C. Then membranes were washed and incubated with secondary horseradish peroxidase-conjugated antibodies for 2 h at room temperature, and the signals were detected using an ECL advanced system (GE Healthcare). Intensity values expressed as the relative protein expression were normalized to β-actin. Primary antibodies used were anti-CD68 (ab955, Abcam); anti-F4/80 (ab6640, Abcam); anti-CD206 (ab64693, Abcam); anti-actinin 4 (15145, Cell Signaling Technology); anti-integrin α4 (8440, Cell Signaling Technology); anti-integrin β1 (4706, Cell Signaling Technology); anti-integrin αL (ab186873, Abcam); anti-integrin β2 (ab185723, Abcam); anti-NF-κB p65 (8242, Cell Signaling Technology); anti-NF-κB p-p65 (3033, Cell Signaling Technology); anti-GR (12041, Cell Signaling Technology); anti-β-actin (ab8226, Abcam). Secondary HRP-conjugated antibodies used were anti-mouse IgG, anti-rabbit IgG, and anti-rat IgG (Abcam).
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