anti-Vimentin, VIM 3B4 + positive western blot control
The set contains enough Western blot control to perform 5 Western blots.
The anti-Vimentin, VIM 3B4 antibody is an excellent marker for mesenchymal cells and mesenchyme-derived tumors (sarcoma, lymphoma, melanoma). The antibody is highly specific for the intermediate filament protein vimentin present in all cells of mesenchymal origin.
|Quantity||50 µg anti-Vimentin + 50 µg western blot control|
|Application||ELISA, ICC, IHC, WB|
|Purification||Affinity chromatography, Cell culture lysate|
|Reactivity||Amphibia, Bovine, Chicken, Human, Monkey|
|Storage||Antibody: short term at 2-8°C; long term in aliquots at -20°C; western blot control: lyophilized at 2-8°C, reconstituted at -20°C; avoid freeze/thaw cycles for both components|
|Intended use||Research use only|
|Immunogen||Vimentin purified from bovine lens|
|Formulation||Antibody: lyophilized; reconstitute in 1 ml dist. water (final solution contains 0.09% sodium azide, 0.5% BSA in PBS buffer, pH 7.4); western blot control: lyophilized, reconstitute in 50 µl 1x SDS buffer (lysis buffer composition: PBS + Pefablock)|
|UniprotID||P48616 (Bovine),P09654 (Chicken),P08670 (Human),V9HWE1 (Human)|
|Tested applications||Tested dilutions|
|Immunocytochemistry (ICC)/ Immunofluorescence (IF)||anti-Vimentin: assay dependent|
|Immunohistochemistry (IHC) - frozen||anti-Vimentin: 1:100-1:200|
|Immunohistochemistry (IHC) - paraffin||anti-Vimentin: 1:100-1:200 (microwave treatment recommended)|
|ELISA||anti-Vimentin: assay dependent|
|Western Blot (WB)||anti-Vimentin: 1:500; western blot control: 10 µg total protein per lane|
The antibody is highly specific for the intermediate filament protein vimentin which is present in all cells of mesenchymal origin. VIM 3B4 has turned out to be the most avid mab to vimentin.
Polypeptide reacting: Mr 57 000 intermediate filament protein (vimentin) of mesenchymal cells.
Tumors specifically detected: sarcoma (including myosarcoma), lymphoma, melanoma.
The binding region of monoclonal antibody VIM3B4 has been characterized by Bohn et al.(1992). According to these authors, the epitope has been localized on the alpha-helical part of vimentin (rod domain coil 2). Due to an aa substitution at position of aa 353 in murine vimentin (that could explain for the weak cross-reaction of the antibody with murine vimentin) they were able to narrow down the binding region around position 353. These findings were confirmed by truncation mutagenesis experiments using human vimentin (Rogers et al., 1995).
Bohn W, Wiegers W, Beuttenmueller M, Traub P: Species-specific recognition patterns of monoclonal antibodies directed against vimentin. Exp Cell Res 201: 1-7 (1992).
Rogers KR, Eckelt A, Nimmrich V, Janssen K-P, Schliwa M, Herrmann H, Franke WW: Truncation mutagenesis of the non-alpha-helical carboxyterminal tail domain of vimentin reveals contributions to cellular localization but not to filament assembly. Eur J Cell Biol 66: 136-150 (1995).
Positive western blot control:
Whole Cell Lysate from SV80 Human Lung fibroblast (SV40-transformed) as positive western blot control.
SV80 whole cell lysate was prepared by homogenization in PBS containing Pefablock.
Protein concentration was determined using Bradford assay.
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- Whole cell lysate from SV80 human lung fibroblast (SV40-transformed) cell line
- Validated and tested western blot control for: anti-Vimentin mouse monoclonal antibody, VIM 3B4, Cat. No. 61013
- Purified, lyophilized
- Mouse monoclonal
- Suitable for ELISA, ICC/IF, IHC and WB
- Reacts with amphibia, bovine, chicken, human and monkey
- Isotype: IgG2a