Human/Mouse/Rat Vimentin Biotinylated Antibody

Catalog #: BAM2105 Datasheet / COA / SDS

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BAM2105

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Human/Mouse/Rat Vimentin Biotinylated Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Vimentin antibodies are ideal for immunocytochemistry colocalization studies in intermediate filaments. The unconjugated antibody detects human Vimentin in Western blots.

Source

Monoclonal Rat IgG_2A Clone # 280618

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human Vimentin
Ser2-Glu466
Accession # P08670

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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Vimentin in HeLa Human Cell Line. Vimentin was detected in formaldehyde fixed HeLa human cervical epithelial carcinoma cell line using Rat Anti-Human/Mouse/Rat Vimentin Biotinylated Monoclonal Antibody (Catalog # BAM2105) at 8 µg/mL overnight at 4 ° C. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (orange; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to intermediate filaments. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Vimentin

Vimentin is a 57 kDa class III intermediate filament (IF) protein that belongs to the intermediate filament family. It is the predominant IF in cells of mesenchymal origin such as vascular endothelium and blood cells (1-3). The human Vimentin cDNA encodes a 466 amino acid (aa) protein that contains head and tail regions with multiple regulatory Ser/Thr phosphorylation sites, and a central rod domain with three coiled-coil regions separated by linkers (1, 2). Human Vimentin shares 97-98% aa identity with mouse, rat, ovine, bovine, and canine Vimentin. Sixteen Vimentin coiled-coil dimers self-assemble to form intermediate (10-12 nm wide) filaments (4). These filaments then anneal longitudinally to form non-polarized fibers that support cell structure and withstand stress (4). IF fibers are highly dynamic, and half-life depends on the balance between kinase and phosphatase activity. For example, phosphorylation followed by dephosphorylation drives IF disintegration, followed by reorganization during mitosis (1, 5, 6). Interactions of head and tail domains link IFs with other structures such as actin and microtubule cytoskeletons (7). Vimentin is involved in positioning autophagosomes, lysosomes and the Golgi complex within the cell (8). It facilitates cell migration and motility by recycling internalized trailing edge integrins back to the cell surface at the leading edge (9-11). Vimentin helps maintain the lipid composition of cellular membranes, and caspase cleavage of Vimentin is a key event in apoptosis (8, 12). Phosphorylation promotes secretion of Vimentin by TNF-alpha -stimulated macrophages (13). Extracellular Vimentin has been shown to associate with several microbes, and appears to promote an antimicrobial oxidative burst (13, 14). Cell-associated Vimentin can also interact with NKp46 to recruit NK cells to tuberculosis-infected monocytes (15).

References