Anti-Vimentin Purified

• Catalog Number: orb44570
• Category: Antibodies
• Description: Mouse monoclonal antibody to Vimentin
• Clonality: Monoclonal
• Clone Number: VI-01
• Tested applications: FC, ICC, IHC-P, WB
• Reactivity: Mammal
• Isotype: Mouse IgM
• Immunogen: Pellet of porcine brain cold stable proteins after depolymerization of microtubules.
• Concentration: 1 mg/ml
• Dilution range: Immunocytochemistry: Staining technique: (a) fix cells for 10 min in methanol at -20°C and for 6 min in acetone at -20°C; (b) fix cells directly in methanol for 10 min at -20°C or in acetone for 10 min at -20°C. Positive control: 3T3 murine Swiss albino fibroblast cell line, RBL rat basophilic leukemia cell line. Flow cytometry: Recommended dilution: 1-5 µg/ml. Intracellular staining. Western blotting: Recommended dilution: 1 µg/ml.
• Purity: Purified by sequential steps of physicochemical fractionation (differential precipitation and solid-phase chromatography methods).
• Conjugation: Unconjugated
• Target: Vimentin
• Entrez: 7431
• UniProt ID: P08670
• RRID: AB_10997622
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: Tris buffered saline (TBS), pH 8.0, 15 mM sodium azide
• Alternative names: anti CTRCT30 antibody, anti Epididymis luminal pro
• Note: For research use only
• Application notes: Immunocytochemistry: Staining technique: (a) fix cells for 10 min in methanol at -20°C and for 6 min in acetone at -20°C; (b) fix cells directly in methanol for 10 min at -20°C or in acetone for 10 min at -20°C. Positive control: 3T3 murine Swiss albino fibroblast cell line, RBL rat basophilic leukemia cell line.
• Expiration Date: 12 months from date of receipt.

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.