Vimentin Antibody

Immunoprecipitation of vimentin from HeLa cell lysate by antibody VI-10 and its detection by antibody VI-01. IgM heavy chain (76-92 kDa) and IgM light chain (25-30 kDa) indicated. Mr of vimentin is 57 kDa. Lr = lysate (reducing conditions); Lnr = lysate (non-reducing conditions); IPr = immunoprecipitate (reducing conditions); IPnr = immunoprecipitate (non-reducing conditions).

Separation of HeLa cells stained using anti-Vimentin (VI-01) purified antibody (concentration in sample 5 µg/ml, GAM APC, red-filled) from HeLa cells unstained by primary antibody (GAM APC, black-dashed) in flow cytometry analysis (intracellular staining of methanol permeabilisated cells).

Immunocytochemistry analysis of vimentin in methanol/acetone fixed murine 3T3 cells using mouse monoclonal antibody VI-01.

Immunohistochemistry staining (paraffin sections) of vimentin in human liver using mouse monoclonal antibody VI-01 (orb44570, diluted 1:400), detected with GAM IgM-Alexa Fluor®488 (diluted 1:200; green), cell nuclei stained with PI (1 µg/ml; orange).

Anti-Vimentin Purified (VI-01) works in WB application under both reducing and non-reducing conditions. Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of JAR, JEG3, HTr-8/SVneo, and HeLa cell lines mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) or non-reducing SDS-loading buffer. Samples were resolved using 10% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed simultaneously with mouse IgM monoclonal antibody VI-01 (1 µg/ml), and mouse IgG1 anti-tubulin monoclonal antibody TU-01 (1 µg/ml) used as the loading control. Subclass-specific secondary antibodies IRDye 800CW Goat-anti-Mouse IgG (green) and IRDye 680RD Goat-anti-Mouse IgM (red) were used for multiplex fluorescent Western blot detection. Vimentin was detected at ~55 kDa.