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Alexa Fluor® 488 anti-Vimentin Antibody

Pricing & Availability
Clone
O91D3 (See other available formats)
Regulatory Status
RUO
Other Names
CTRCT30, HEL 113, Epididymis Luminal Protein 113
Isotype
Mouse IgG2a, κ
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Product Citations
publications
O91D3_A488_Vimentin_Antibody_102720
HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 0.5 µg/ml Alexa Fluor® 488 anti-Vimentin (clone O91D3) or Alexa Fluor® 488 Mouse IgG2a, κ Isotype Ctrl Antibody overnight at 4 degree. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
  • O91D3_A488_Vimentin_Antibody_102720
    HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with 0.5 µg/ml Alexa Fluor® 488 anti-Vimentin (clone O91D3) or Alexa Fluor® 488 Mouse IgG2a, κ Isotype Ctrl Antibody overnight at 4 degree. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
  • O91D3_A488_Vimentin_Antibody_2_031622
    Jurkat cells (filled histogram, positive control) and Daudi cells (open histogram, negative control) were fixed with Fixation Buffer (Cat. No. 420801), permeabilized using True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with Alexa Fluor® 488 anti-Vimentin (clone O91D3).
  • O91D3_A488_Vimentin_Antibody_3_031324
    IHC staining of Alexa Fluor® 488 anti-Vimentin (clone O91D3) on formalin-fixed paraffin-embedded human tonsil. Following antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902), the tissue was incubated with 5 µg/mL of Alexa Fluor® 488 anti-Vimentin (clone O91D3) (green) and co-stained with Alexa Fluor® 594 anti-Pan-Cytokeratin (clone AE-1/AE-3) (Cat. No. 914206) (magenta). Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured with a 10X objective. Scale bar: 50 µm
  • O91D3_A488_Vimentin_Antibody_4_101024
    Multiplexed IHC staining of Alexa Fluor® 488 anti-Vimentin (clone O91D3) on formalin-fixed paraffin-embedded human tonsil tissue, validated for use on the Canopy Cellscape™. The tissue was iteratively stained with Alexa Fluor® 488 anti-Vimentin (clone O91D3, green) and Alexa Fluor® 488 anti-CD3 (red) for one hour at room temperature. Nuclei were counterstained with DAPI. Images were captured with a 20X objective. Scale bar: 50 µm
Compare all formats See Alexa Fluor® 488 spectral data
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677809 25 µg £94
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Description

Vimentin are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is a widely expressed and highly conserved 54 kD protein that is constitutively expressed in mesenchymal cells, endothelial cells lining blood vessels, renal tubular cells, macrophages, neutrophils, fibroblasts, and leukocytes1,2. Vimentin is used as a marker of mesenchymal cells to distinguish them from epithelial cells3. Increased vimentin expression is frequently used as an EMT marker in cancer4. Autoantibodies to vimentin are commonly found in patients with autoimmune diseases such as Lupus5 and rheumatoid arthritis6, and also found after transplantation7.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Full length human vimentin produced in E. coli.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICC - Quality tested
ICFC, IHC-P - Verified
SB - Community Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, the suggested use of this reagent is 0.2 - 2 µg per mL. For intracellular flow cytometry using our True-Phos™ Perm Buffer, the suggested use of this reagent is ≤ 1.0 µg per million cells in 100 µL volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Blue Laser (488 nm)
Application Notes

While this clone recognizes mouse Vimentin, we do not recommend its usage for western blot due to poor affinity of the antibody for the protein. Additional reported applications for the relevant formats include: spatial biology (IBEX)1,2.

Additional Product Notes

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  2. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Cherkaoui S, et al. 2022. Mol Syst Biol. 18:e11033. PubMed
  2. Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed
  3. Buljan M, et al. 2020. Molecular Cell. 79(3):504-520. PubMed
  4. Nethisinghe S, et al. 2021. Int J Mol Sci. 22:. PubMed
  5. Rodríguez E, et al. 2022. Commun Biol. 5:41. PubMed
  6. Zhou WJ, et al. 2022. Sci Adv. 8:eabj2488. PubMed
RRID
AB_2650955 (BioLegend Cat. No. 677809)

Antigen Details

Structure
466 amino acids with a predicted molecular weight of approximately 54 kD.
Distribution

Cytoplasm.

Function
Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
Interaction
HCV core protein, LGSN, SYNM, PLEC, SLC6A4, STK33, LARP6, RAB8B, TOR1A, TOR1AIP1, and BCAS3.
Cell Type
B cells, Mesenchymal Stem Cells, Neural Stem Cells, Neutrophils
Biology Area
Cell Adhesion, Cell Biology, Cell Motility/Cytoskeleton/Structure, Immunology, Neuroscience, Neuroscience Cell Markers, Stem Cells
Molecular Family
Intermediate Filaments
Antigen References

1. Kidd ME, et al. 2014. Am. J. Respir. Cell Mol. Biol. 50:1.
2. Fuchs E, et al. 1994. Annu. Rev. Biochem. 63:345.
3. Zeisberg M, et al. 2009. J. Clin. Invest. 119:1429.
4. Scanlon CS, et al. 2013. J. Dent. Res. 92:114.
5. Thebault S, et al. 2002. J. Immunol. 169:4046.
6. Vossenaar ER, et al. 2004. Arthritis Res. Ther. 6:R142.
7. Rose ML. 2013. Hum. Immunol. 74:1459.

Gene ID
7431 View all products for this Gene ID
UniProt
View information about Vimentin on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Go To Top Version: 6    Revision Date: 10/10/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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