VIM Monoclonal Antibody
Description
Vimentin Biology
Vimentin is a 57–58 kDa type III intermediate filament protein expressed in mesenchymal cells, including fibroblasts, endothelial cells, and lymphocytes. It stabilizes cytoskeletal interactions, regulates cell migration, and provides mechanical resilience to cellular stress . Overexpression is linked to epithelial-to-mesenchymal transition (EMT) in cancers, such as melanoma, sarcoma, and lymphoma .
| Property | Details |
|---|---|
| Function | Maintains cytoskeletal structure, regulates cell migration, and EMT |
| Expression | Mesenchymal cells, activated macrophages, metastatic cancer cells |
| Molecular Weight | 57–58 kDa |
| Key Domains | N-terminal head, rod domains (coils 1A, 1B, 2A, 2B), C-terminal tail |
Key VIM Antibody Clones
Monoclonal antibodies differ in epitope recognition, cross-reactivity, and applications:
Epitope Determination
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V9: Recognizes a 13-amino acid sequence (aa 411–423) in the C-terminal tail. Asparagine at position 417 is critical for binding .
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VIM 3B4: Targets the α-helical region (rod domain coil 2), with species-specific reactivity due to amino acid substitutions (e.g., murine vimentin) .
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VM1170: Binds full-length human VIM with no cross-reactivity to desmin, keratin, or neurofilaments .
Technical Uses
Diagnostic Utility
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Tumor Classification: VIM antibodies aid in diagnosing sarcoma, melanoma, and lymphoma by excluding epithelial markers like keratin .
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EMT Detection: Overexpression correlates with aggressive metastatic phenotypes in breast, lung, and oral cancers .
Species Reactivity
Interference Risks
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Non-Specific Binding: Some ascites-derived antibodies (e.g., SAB4200716) may require blocking steps to reduce background .
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Epitope Masking: Paraffin embedding or protease treatments may alter epitope accessibility, necessitating optimization .
Emerging Applications
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Targeted Therapies: Antibodies like hzVSF block vimentin-mediated viral entry (e.g., HBV) by targeting surface-exposed vimentin on infected cells .
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Biomarker Development: Serum vimentin levels correlate with liver and colon cancer progression, prompting ELISA-based screening tools .
Challenges
Product Specs
This product is a mouse monoclonal antibody generated against the recombinant protein corresponding to the full length of human vimentin. The VIM antibody is an unconjugated IgG isoform. Its quality has been rigorously validated through ELISA and WB testing. This antibody demonstrates reactivity with human, mouse, and rat samples. Importantly, it exhibits no cross-reactivity with other closely related intermediate filament (IF) proteins, such as keratin. The VIM antibody specifically binds to vimentin, a mesenchymal cell-expressed IF protein that plays a crucial role in maintaining cellular integrity, stabilizing cytoskeletal interactions, and providing resistance to cellular damage.
Target Background
Vimentins are class-III intermediate filaments found in various non-epithelial cells, particularly mesenchymal cells. Vimentin is associated with the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally. It collaborates with LARP6 in stabilizing type I collagen mRNAs for CO1A1 and CO1A2.
- This study investigated the impact of vimentin expression on the mechano-elastic and migratory properties of the highly invasive breast carcinoma cell line MDA231. Results demonstrated that vimentin stiffens cells and enhances cell migration in dense cultures, but exerts minimal or no effect on the migration of sparsely plated cells. PMID: 29022351
- Positive vimentin expression has been identified as a potential poor prognostic marker in gastric cancer (review, meta-analysis). PMID: 30078472
- High vimentin expression is associated with pancreatic cancer. PMID: 29956814
- HDAC inhibitors have been observed to augment both E-cadherin and vimentin expression, with their effects varying across different cholangiocarcinoma cell lines. Consequently, the clinical application of HDAC inhibitors in biliary cancer should be approached with caution. PMID: 29767267
- miR-373 has been shown to suppress gastric cancer metastasis by downregulating vimentin. PMID: 29257346
- Vimentin plays a role in the M2BP inhibition of HIV-1 virion production. M2BP mediates the interaction between HIV-1 Gag and Vimentin. PMID: 27604950
- Desmin, Glial Fibrillary Acidic Protein, Vimentin, and Peripherin are type III intermediate filaments that have roles in both health and disease (review). PMID: 29196434
- Silencing Vimentin in CNE2 cells leads to a reduction in microvessel density and expressions of VEGF, CD31, MMP2, and MMP9 in pulmonary metastatic tumors. PMID: 28744809
- A carcinoid-like/labyrinthine pattern of cell arrangement in vimentin/cytokeratin 20 expressing sebaceous neoplasms may represent a morphological phenotype of sebaceous mantles. PMID: 28027080
- Cell surface vimentin facilitates DENV-2 infection of vascular endothelial cells. PMID: 27910934
- HIF-1alpha expression was upregulated in the vasculogenic mimicry-positive CRC cell line HCT-116, influencing the expression of EMT-related markers Claudin-4, E-cadherin (E-cd), and Vimentin(VIM). PMID: 27869227
- Research findings indicate that vimentin in human GC tissues and cell lines is upregulated due to its de-ubiquitination after interactions with USP14 and miR-320a, potentially contributing to the aggressive nature of GC cells. PMID: 27448976
- Knocking down long pentraxin-3 (PTX3) or vimentin repressed oleate-induced head and neck squamous cell carcinomas (HNSCCs) invasion. PMID: 28489600
- Stromal vimentin expression emerges as a promising indicator for survival prediction and adjuvant chemotherapy response in patients with stage II colorectal cancer exhibiting high-risk factors for recurrence. PMID: 28611349
- Vimentin and its interaction with Shigella flexneri IpaC are not essential for effector translocation pore formation but are required for stable docking of Shigella flexneri to cells. Notably, stable docking triggers effector secretion. PMID: 27572444
- This study reports the elongation reaction of vimentin in both solution and in situ using time-resolved static and dynamic light scattering. PMID: 27655889
- Vimentin induced by exosomes is crucial for lung cancer to induce mesenchymal transition (EMT) in recipient bronchial epithelial cells (HBECs). PMID: 27363026
- Results demonstrate that vimentin silencing in ovarian cancer cells upregulates proteins involved in the exocytotic process, leading to a decrease in cellular cisplatin accumulation. PMID: 27322682
- The combined biomarkers E-cadherin, membranous epidermal growth factor receptor (EGFR), and vimentin exhibit stronger prognostic value for disease-free survival than any of the individual biomarkers. PMID: 27172790
- An AHR protein-vimentin protein complex is formed in the cytoplasm, resulting in proteasome degradation of vimentin. PMID: 27752740
- TRIM56 is the ubiquitin ligase that degrades vimentin in ovarian cancer cells, regulating cell migration and neoplasm invasiveness. PMID: 28771721
- TIS21 attenuates Doxorubicin-induced cancer cell senescence by inhibiting linear actin nucleation via the Nox4-ROS-ABI2-DRF signaling cascade. PMID: 27932314
- Findings suggest a mechanism whereby OPN induces epithelial-mesenchymal transition of hepatocellular carcinoma cells through increasing vimentin stability. PMID: 26824421
- Research indicates that miR-375 suppresses RUNX1 levels, leading to reduced vimentin and L-plastin expression. Knockdown of RUNX1, L-plastin, and vimentin resulted in significant reductions in cell invasion in vitro, highlighting the functional significance of miR-375 regulation of specific proteins involved in head and neck squamous cell carcinoma (HNSCC) invasion. PMID: 28499703
- Vimentin expression was identified as an adverse prognostic factor for DSS in TSCC patients, even after adjusting for cell differentiation, pathological stage, and expression levels of Snail, Twist, E-cadherin, and N-cadherin. Snail, E-cadherin, N-cadherin, and Vimentin were associated with tumorigenesis and pathological outcomes. PMID: 28570699
- Circulating anti-vimentin IgG autoantibody levels are significantly higher in idiopathic pulmonary fibrosis subjects compared to normal controls. PMID: 28754682
- The RhoA/ROCK and Raf-1/CK2 pathways are responsible for TNF-alpha-mediated endothelial cytotoxicity through regulation of the vimentin cytoskeleton. PMID: 28743511
- Findings suggest that islet cell expression of vimentin indicates a degree of plasticity and dedifferentiation with potential loss of cellular identity in diabetes. PMID: 28348116
- Research has identified a role for members of specific signaling pathways in the regulation of EGF-induced vimentin expression in the MDA-MB-468 breast cancer cell line. PMID: 27163529
- Changes in methylation levels in cfDNA are associated with hepatocellular carcinoma and could represent useful plasma-based biomarkers. PMID: 28333958
- Vimentin regulates the differentiation switch through modulation of K5/K14 expression. Moreover, due to a significant correlation between high vimentin-K14 expression and recurrence/poor survival in oral cancer patients, vimentin-K14 together may prove to be novel markers for prognostication in human oral cancer. PMID: 28225793
- Data indicates that ellagic acid (EA) down-regulates the expression of COX-2, NF-kappa B, vimentin, and up-regulates the expression of E-cadherin in pancreatic carcinoma PANC-1 cells. PMID: 28135203
- Bevacizumab treatment has been linked to structural protein abnormalities, with decreased GFAP and vimentin content and upregulated GFAP and vimentin mRNA expression. PMID: 28419863
- Reducing cell surface vimentin through small interfering RNA (siRNA) knockdown in HeLa and NIKS cells significantly increased human papillomavirus 16 infectious internalization, while vimentin overexpression had the opposite effect, identifying vimentin as a viral restriction factor. PMID: 28566373
- Data suggests that filamentous vimentin beneath the plasma membrane is involved in enhancing integrin adhesiveness, implying that regulating the vimentin-integrin interaction could potentially control cell adhesion. PMID: 27044755
- Vimentin depletion induces phosphorylation of the microtubule-associated GEF-H1 on Ser886, promoting RhoA activity and actin stress fiber assembly. PMID: 28096473
- Data shows that the filament elongation of both desmin and keratin K8/K18 proceeds very similarly to that of vimentin. PMID: 27304995
- The coexistence of vimentin-positive and Axl-high expression is a poor prognostic factor for primary breast cancer. Vimentin and Axl expression may contribute to the aggressive phenotype in breast cancer. PMID: 27506606
- Findings suggest that the HIF-1alpha-HDAC1 complex transcriptionally inhibits miR-548an expression during hypoxia, resulting in the upregulation of vimentin that facilitates pancreatic tumorigenesis. PMID: 27353169
- Rab7a depletion decreases the amount of active Rac1 but not its abundance and reduces the number of cells with vimentin filaments facing the wound, indicating that Rab7a plays a role in the orientation of vimentin filaments during migration. PMID: 27888097
- Over-expression of TNC, SMA, and vimentin were significantly correlated with lower overall survival in prostate cancer patients. PMID: 28341124
- Data demonstrates that VIM mRNA is regulated in melanoma by UNR protein. PMID: 27908735
- These findings suggest that Plk1 regulates smooth muscle contraction by modulating vimentin phosphorylation at Ser-56. PMID: 27662907
- Structural Dynamics of the Vimentin Coiled-coil Contact Regions Involved in Filament Assembly as Revealed by Hydrogen-Deuterium Exchange. ( PMID: 27694444
- This study provides the first evidence demonstrating that vimentin is critical for influenza viral infection as it facilitates endosomal trafficking and acidification, and mediates viral genome penetration into the cytoplasm to propagate the infection. PMID: 27423069
- RAP1 promotes colorectal cell migration through the regulation of Vimentin, suggesting that RAP1 may serve as a potential target for the diagnosis and therapy of CRC. PMID: 28381157
- High vimentin expression is associated with Non Small-cell Lung Cancer. PMID: 28373440
- Data demonstrates that keratinocyte migration requires the interaction between vimentin and keratins at the -YRKLLEGEE- sequence at the helical 2B domain of vimentin. PMID: 27072292
- High vimentin expression is associated with malignant pleural mesothelioma. PMID: 27646775
- Increased TET1 induced re-expression of vimentin through active DNA demethylation, causing partial epithelial-to-mesenchymal (EMT) in A2780 cells. PMID: 28150354
Q&A
What is the biological significance of Vimentin in cellular functions?
Vimentin serves multiple critical functions in cellular biology. It attaches to the nucleus, endoplasmic reticulum, and mitochondria, contributing to cell structural integrity. Research indicates that vimentin plays crucial roles in cell directional movement, orientation, and cell sheet organization. It also contributes to Golgi complex polarization at the cell migration front . Additionally, vimentin protects SCRIB from proteasomal degradation and helps localize it to intermediate filaments in a cell contact-mediated manner. It works with LARP6 in stabilizing type I collagen mRNAs for CO1A1 and CO1A2 . Understanding these functions is important when selecting vimentin as a target in your experimental system.
Which applications are most suitable for VIM monoclonal antibodies?
VIM monoclonal antibodies are versatile tools applicable across multiple laboratory techniques:
| Application | Typical Working Dilutions | Notes |
|---|---|---|
| Western Blotting | 0.01-2 μg/mL | Detects ~58kDa protein band |
| Immunohistochemistry | 5-20 μg/mL | Works on paraffin-embedded tissues |
| Immunocytochemistry/Immunofluorescence | 5-40 μg/mL | Excellent for filament visualization |
| Flow Cytometry | Optimal dilution should be experimentally determined | For intracellular staining |
For reliable results, it's recommended to optimize the antibody concentration for your specific experimental conditions and cell/tissue types . When using in flow cytometry, proper cell permeabilization is essential since vimentin is an intracellular protein .
How do I select the appropriate VIM monoclonal antibody clone for my research?
Selection of the appropriate VIM antibody clone depends on your experimental goals, target species, and application:
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Target epitope consideration: Different clones recognize different domains of vimentin. For example, mAb 3B4 targets the rod domain, while mAb V9 recognizes the tail domain .
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Species reactivity: Check the cross-reactivity profile of the antibody. For example, clone VI-RE/1 reacts with human samples , while clone VIM/1937R reacts with human, rat, porcine, bovine, canine, chicken, equine, and feline samples but not with mouse samples .
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Application compatibility: Verify that the clone has been validated for your specific application. Some clones perform better in certain applications than others .
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Conjugation needs: Determine if you need an unconjugated antibody or one conjugated to a fluorophore (like mFluor Violet 610) based on your detection method .
What controls should I include when using VIM monoclonal antibodies?
To ensure experimental validity:
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Positive control: Use cell lines or tissues known to express vimentin such as fibroblasts, endothelial cells, or mesenchymal cells. Melanoma cell lines are also strongly vimentin-positive .
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Negative control: Include hepatoma or seminoma samples, which are among the few tumor types that do not express vimentin . Mouse samples would serve as negative controls specifically for the VIM/1937R clone .
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Isotype control: Include an antibody of the same isotype (e.g., IgG1 or IgG2b) but irrelevant specificity to control for non-specific binding .
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Procedural control: Perform the experiment without the primary antibody to evaluate secondary antibody specificity and background signal levels.
How does epitope specificity affect the performance of different VIM monoclonal antibody clones?
The epitope specificity of VIM monoclonal antibodies significantly impacts their performance across different applications and research contexts. The V9 clone, one of the most widely used vimentin antibodies, recognizes a specific thirteen amino acid sequence (411ISLPLPNFSSLNL423) in the C-terminal region of human vimentin . Within this epitope, the asparagine residue at position 417 has been identified as critical for antibody binding .
This epitope specificity has several implications:
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Cross-reactivity patterns: Antibodies recognizing highly conserved epitopes will demonstrate broader cross-species reactivity. Comparing amino acid sequence alignments of vimentin across species can predict cross-reactivity .
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Application performance: The location of the epitope can affect antibody performance in different applications. C-terminal targeting antibodies like V9 may perform differently in applications where protein folding or fragmentation occurs compared to those targeting other domains .
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Sensitivity to post-translational modifications: If the epitope contains sites for phosphorylation, glycosylation, or other modifications, antibody binding may be affected by the modification status of vimentin in your sample .
Understanding the precise epitope of your chosen antibody can help explain inconsistent results across different experimental conditions or sample types.
What strategies should be employed to resolve false negative results with VIM monoclonal antibodies in immunohistochemistry?
False negative results in vimentin immunostaining may occur for several methodological reasons:
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Epitope masking: The formalin fixation process can mask epitopes through protein cross-linking. Implement optimized antigen retrieval methods:
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Antibody concentration: Titrate the antibody to determine optimal concentration. For vimentin antibodies, concentrations between 5-20 μg/mL are typically effective for IHC .
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Detection system sensitivity: Consider amplification systems like tyramide signal amplification (TSA) for weakly expressed vimentin.
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Sample preservation issues: Vimentin is relatively sensitive to prolonged storage. Use freshly cut sections when possible and store unstained slides properly .
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Alternative clone selection: If a particular clone fails consistently, try an alternative clone targeting a different epitope. For example, if the V9 clone (C-terminal specific) yields negative results, try a clone targeting the rod domain .
How can VIM monoclonal antibodies be utilized in multiplex immunostaining protocols?
Multiplex immunostaining with vimentin antibodies requires careful planning:
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Compatible antibody selection:
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Panel design for tumor classification:
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Technical optimization:
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Controls for multiplex validation:
What are the potential pitfalls in quantitative analysis of VIM expression using monoclonal antibodies?
Quantitative analysis of vimentin expression presents several challenges:
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Epitope availability variations:
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Antibody saturation considerations:
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Expression heterogeneity:
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Reference standards:
How does cross-species reactivity of VIM monoclonal antibodies impact experimental design in comparative studies?
Cross-species reactivity considerations are crucial when designing comparative studies:
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Epitope conservation analysis:
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Species-specific validation:
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Experimental design recommendations:
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Alternative approaches:
