The protein levels of CBS and CSE in the kidney were measured by Western immunoblotting analysis. In brief, kidney proteins (20 μg) were separated by electrophoresis in 10% SDS polyacrylamide gels. Proteins in the gel were transferred to a nitrocellulose membrane. The membrane was probed with (Abe and Kimura 1996 (link)) mouse anti‐CBS monoclonal (1:3,000; Abnova Corporation, Taipei, Taiwan) or rabbit anti‐CSE monoclonal antibodies (1:3,000; GeneTex, Irvine, CA) for rat proteins. HRP‐conjugated anti‐mouse or anti‐rabbit IgG antibodies (Cell Signaling Technology, Danvers, MA) were used as the secondary antibodies (1:5,000). The corresponding protein bands were visualized using enhanced chemiluminescence reagents and analyzed with a gel documentation system (Bio‐Rad Gel Doc1000). To confirm the equal loading of proteins for each sample, the same membranes were reprobed with mouse anti‐β‐actin monoclonal antibodies (1:5,000, Cell Signaling Technology). Proinflammatory factors (MCP‐1, IL‐6) in the kidney and plasma as well as plasma neutrophil gelatinase‐associated lipocalin (NGAL) were measured using the MesoScale Discovery electrochemiluminescence platform (Rockville, MD).
Mouse anti cbs monoclonal
Manufactured by Abnova
Sourced in Taiwan, Province of China
Mouse anti‐CBS monoclonal is a laboratory reagent for the detection and analysis of the cystathionine beta-synthase (CBS) protein. It is a monoclonal antibody produced in mice that specifically binds to the CBS protein. This product can be used in various immunoassay applications to identify and quantify the presence of CBS in biological samples.
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Lab products found in correlation
Gel doc 1000, by Bio-Rad (1 mentions)
Anti β actin mouse monoclonal antibody, by Cell Signaling Technology (1 mentions)
Hrp conjugated anti mouse or anti rabbit igg antibodies, by Cell Signaling Technology (1 mentions)
Fv1000 microscope, by Olympus (1 mentions)
Eight chamber cell culture slides, by BD (1 mentions)
Atto 488, by Merck Group (1 mentions)
2 protocols using mouse anti cbs monoclonal
1
Quantification of CBS and CSE Proteins in Kidney
2
Immunofluorescence Localization of CBS
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The A23 CBS WT, I278T or R125Q cells were grown in a complete MEM media on eight-chamber cell culture slides (Falcon) to 70% confluency. Cells were then fixed with 4% paraformaldehyde for 10 min and permeabilized by methanol. For visualization and localization of desired targets, we used the following antibodies (dilution, manufacturer): mouse anti-CBS monoclonal (1,000×, Abnova) or rabbit anti-CBS polyclonal (2,500×, lab-made), rabbit anti-BiP monoclonal (200×, Cell Signaling), rabbit anti-HSP70 polyclonal (200×, Cell Signaling) and mouse anti-20S proteosome monoclonal (200×, Enzo Life Sciences). Appropriate anti-mouse and rabbit IgG secondary antibodies conjugated to either Atto 488 (Sigma) or Alexa Fluor 647 fluorophores (Molecular Probes) were used for immunostaining, while DAPI Fluoromont chemical stain (SouthernBiotech) was used to visualize nuclei. Slides were analyzed using Olympus FV-1000 microscope with Fluoview software for image acquisition. Images were normalized to background fluorescence, quantified using ImageJ package and analyzed using GraphPad Prism software. Statistical significance was designated by asterisks as follows: *P < 0.05, **P < 0.01, and ***P < 0.001.
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