Vimentin Mouse Monoclonal Antibody [A6-C1]
cat.: M1412-1
| Product Type: | Mouse monoclonal IgG2b, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | A6-C1 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 2ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 54 kDa |
| Isotype: | IgG2b |
| Immunogen: | Synthetic peptide within Human Vimentin aa 1-50 / 466. |
| Positive control: | HeLa cell lysate, C2C12 cell lysate, L6 cell lysate, HeLa, human kidney tissue. |
| Subcellular location: | Cytoplasm, cytoskeleton, Nucleus matrix, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:2,000 1:100-1:200 1:400 |
| Uniprot #: | SwissProt: P08670 Human | P20152 Mouse | P31000 Rat |
| Alternative names: | CTRCT30 Epididymis luminal protein 113 FLJ36605 HEL113 VIM VIME_HUMAN Vimentin |
Images
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Fig1:
Western blot analysis of Vimentin on different lysates with Mouse anti-Vimentin antibody (M1412-1) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: C2C12 cell lysate Lane 3: L6 cell lysate Lane 4: Daudi cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 54 kDa Exposure time: 24 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1412-1) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
All lanes: Western blot analysis of Vimentin with anti-Vimentin antibody (M1412-1) at 1:500 dilution. Lane 1: Wild-type Hela whole cell lysate (10 µg). Lane 2: Vimentin knockout Hela whole cell lysate (10 µg). M1412-1 was shown to specifically react with Vimentin in wild-type Hela cells. No band was observed when Vimentin knockout sample was tested. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (M1412-1, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Immunocytochemistry analysis of HeLa cells labeling Vimentin with Mouse anti-Vimentin antibody (M1412-1) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Vimentin antibody (M1412-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-Vimentin antibody (M1412-1) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1412-1) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.