WB result of Vimentin Rabbit mAb
Primary antibody: Vimentin Rabbit mAb at 1/500 dilution
Lane 1: Daudi whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: A549 whole cell lysate 20 µg
Negative control: Daudi whole cell lysate
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 54 kDa
Observed MW: 57 kDa
Exposure time: 150 s
Vimentin Recombinant Rabbit mAb (SDT-029-120)
Vimentin Recombinant Rabbit mAb (SDT-029-120)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Vimentin |
| Synonyms | VIM |
| Location | Cytoplasm, Cytoskeleton, Cell membrane |
| Accession | P08670 |
| Clone Number | SDT-029-120 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, IP, ICFCM |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Pr, Bv, Cz, Pg, Hm, Av |
| Purification | Protein A |
| Concentration | 0.25 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:500 | |
| IP | 1:25 | |
| IHC | 1:1000 | |
| ICC | 1:500 |
Background
Vimentin is a type III intermediate filament (IF) protein that is expressed in mesenchymal cells. IF proteins are found in all animal cells as well as bacteria. Intermediate filaments, along with tubulin-based microtubules and actin-based microfilaments, comprises the cytoskeleton. All IF proteins are expressed in a highly developmentally-regulated fashion; vimentin is the major cytoskeletal component of mesenchymal cells. Because of this, vimentin is often used as a marker of mesenchymally-derived cells or cells undergoing an epithelial-to-mesenchymal transition (EMT) during both normal development and metastatic progression. It can be used as a differential diagnosis between epithelial tumors and mesenchymal tumors, as well as for endometrial adenocarcinoma and cervical adenocarcinoma.
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Picture
Western Blot
WB result of Vimentin Rabbit mAb
Primary antibody: Vimentin Rabbit mAb at 1/500 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 54 kDa
Observed MW: 57 kDa
Exposure time: 120 s
WB result of Vimentin Rabbit mAb
Primary antibody: Vimentin Rabbit mAb at 1/500 dilution
Lane 1: rat brain lysate 40 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 54 kDa
Observed MW: 55 kDa
Exposure time: 180 s
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Vimentin antibody at 1/250 dilution (0.1 μg) / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
Vimentin Rabbit mAb at 1/25 dilution (1 µg) immunoprecipitating Vimentin in 0.4 mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using Vimentin Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: HeLa whole cell lysate 20 µg (Input)
Lane 2: Vimentin Rabbit mAb IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 54 kDa
Observed MW: 55 kDa
Exposure time: 60 s
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human kidney. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cervical squamous cell carcinoma. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung cancer. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse colon. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat kidney. Anti-Vimentin antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Vimentin antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
