Mouse anti-Human Vimentin (unconjugated), Clone VIM3B4, IgG2a, Purified

Concentrated Antibody for Immunohistochemistry

SKU:
K024
Category
Quantity
0, 05 mg
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Background

Vimentin is an intermediate filament with a relative molecular mass of 57kD. It forms, together with microfilaments and microtubules, the cytoskeleton of non-epithelial cells, e.g. non-muscle mesenchymal cells and cells of mesenchymal origin including vascular smooth muscle, astrocytes and numerous cultured cell lines. It frequently co-occurs with other intermediate filaments such as desmin.

Expression of this protein is retained during the conversion of normal to neoplastic cells.

Synonyms

Vimentin

Volume/Quantity

0.05 mg

Product Form

Purified IgG – lyophilised

Reconstitution

Reconstituted with 1 ml distilled water

Preparation

Purified IgG prepared by affinity chromatography on Protein A from ascites fluid.

Clone Number

VIM3B4

Buffer

Phosphate buffered saline

Approx. Protein Concentrations

IgG concentration 0.05 mg/ml

Preservatives Stabilisers

0.09 mM Sodium Azide (NaN3)
0.5% Bovine Serum Albumin (BSA)

Immunogen

Vimentin purified from bovine lens

Isotype

IgG2a (Mouse)

Specificity

This antibody is positive with cells of mesenchymal derivation, including endothelial cells and certain smooth muscle cells of the vascular tract, fibroblasts, connective tissue, all types of blood cells, including thymocytes, interstitial and testicular Sertoli cells, ovarian follicle cells. It can also be used to detect coexpression of vimentin in combination with other intermediate filament proteins.
K024 can also stain permanent cell lines such as RD cells, glioma cells, fibroblasts (SV-80, 3T3), BHK. The antibody recognizes all vimentin expressing tumours such as sarcomas (including myosarcomas),
lymphomas, melanomas.

Fusion Partners

Spleen cells from immunised Balb C/J mice were fused with cells of the P3X63Ag8.653 mouse myeloma cell line

Suggested Working Dilution

FlowCytometry Not tested
Immunohistology-frozen Yes 1/100
Immunohistology-paraffin Yes 1/100
Immunohistology-resin Not tested
Immunofluorescence Yes
ELISA Yes
Immunoprecipitation Not tested

Western Blotting Yes
Radioimmunoassay Not tested

Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.

Immunohistology

Pre-treatment

Treatment prior to staining of paraffin sections with 0.1% Pronase (LINARIS CatNo E110) improves the immunostaining

Positive Control Tissue

Positive in RD cells, glioma cells, fibroblasts (SV-80), and MDCK cell line.

Recommended Secondary Reagents

F(ab’)2 rabbit anti-mouse IgG HRP conjugate – (LINARIS CatNo LST0013B)
ABC-Kit Mouse IgG-POD Labelling (LINARIS CatNo EDP4002)
DAB-Substrate for POD (LINARIS CatNo E108)
ABC-Kit Mouse IgG AP Labelling (LINARIS CatNo EDA5002)
BCIP/NBT Substrate for AP (LINARIS CatNo ESA5400)

Recommended Negative Controls

Mouse IgG2a Negative Control (LINARIS CatNo ITC0929)

Westernblotting

Chemiluminescent Substrate

DuoLuX Chemiluminescent Substrate for Alkaline Phosphatase (AP) (LINARIS CatNo ESA6605) and Peroxidase (POD) (LINARIS CatNo ESP6604).

Western Blotting Immunodetection Kit

ABC-AmP Chromogenic Detection Kit anti-Mouse IgG (LINARIS CatNo EDA6402) or anti-Rabbit IgG (LINARIS CatNo EDA6401)

Weight Standard

Biotinylated protein molecular weight marker (LINARIS CatNo RPB1400). Molecular weight range from 19kD to 222kD

Storage Conditions

Prior to reconstitution store at 2-8°C. Reconstituted antibody is stable for at least one month when stored
at 2-8°C. For long term storage, it is recommended that aliquots of the antibody are frozen at –20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.

Shelf Life

12 months from date of reconstitution.

Franke, W. W. & Moll, R. Differentiation 36, 145-63 (1987)

Heid, H. W. et al. Differentiation 37, 137-57 (1988)

Kasper, M. et al. Histochem. 93, 93-103 (1989)

Bohn, W. et al. Exp. Cell Res. 201, 1-7 (1992)

Specificity

Vimentin
Species Reactivity

Amphibien, Bovine, Chicken, Dog, Human, Monkey
Host / Source

Mouse
Isotype

IgG2a
Application

EIA, IF, IHC-F, IHC-P, WB
Clone

VIM3B4
Antigen

Human Vimentin
Quantity

0, 05 mg
Format

Purified
Storage Temperature

2-8 °C / -20 °C
Shipping Temperature

2-8 °C

Documents

pdf Datasheet