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Purified 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
Ave. Rating
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Product Citations
publications
a-O91D3_PURE_Vimentin_Antibody_1_113018
Total cell lysates (15 µg total protein) from Daudi (negative control), PC-3, Jurkat and NIH/3T3 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a nitrocellulose membrane, and probed with 0.25 µg/mL (1:2000 dilution) of Purified anti-Vimentin Antibody, clone O91D3, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular Weight marker. Predicted expression data was obtained from Human Protein Atlas.
  • a-O91D3_PURE_Vimentin_Antibody_1_113018
    Total cell lysates (15 µg total protein) from Daudi (negative control), PC-3, Jurkat and NIH/3T3 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a nitrocellulose membrane, and probed with 0.25 µg/mL (1:2000 dilution) of Purified anti-Vimentin Antibody, clone O91D3, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular Weight marker. Predicted expression data was obtained from Human Protein Atlas.
  • b-O91D3_PURE_Vimentin_Antibody_2_113018
    HeLa cells were stained with purified anti-Vimentin (clone O91D3) antibody, followed by staining with Alexa Fluor® 594 conjugated goat anti-mouse IgG antibody (red). Actin filaments were labeled with Alexa Fluor® 647 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 40X objective.
  • c-O91D3_PURE_Vimentin_ICFC_110121
    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 0.25 µg/test of purified anti-Vimentin antibody (clone O91D3) followed by PE goat anti-mouse IgG antibody (Cat. No. 405307).
  • d-O91D3_PURE_Vimentin_IHC-P_110121
    IHC staining of anti-Vimentin antibody (clone O91D3) on formalin-fixed paraffin-embedded human kidney (A) and brain (B) tissues. Following antigen retrieval using Sodium Citrate H.I.E.R. (Cat. No. 928502), the tissues were incubated with 5 µg/mL of the anti-Vimentin antibody overnight at 4°C. BioLegend’s Ultra Streptavidin HRP Kit (Multi-Species, DAB, Cat. No. 929501) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The images were captured with a 40X objective. Scale bar: 50 μm.
  • e-O91D3_PURE_Vimentin_IHC-F_110121
    IHC staining of anti-Vimentin antibody (clone O91D3) on frozen human spleen (A) and brain (B) tissues. Following fixation with Fixation Buffer (Cat. No. 420801) and permeabilization with 0.5% Triton X-100, the tissue sections were incubated with 5.0 µg/mL of anti-Vimentin antibody overnight at 4°C followed by incubation with Alexa Fluor® 594 Goat anti-mouse IgG antibody (Cat. No. 405326) for 2-hours at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) and the slides were mounted with ProLong™ Gold Antifade Mountant. These images were captured with a 40X objective. Scale bar: 50μm.
  • F_O91D3_PURE_Vimentin_Antibody_111723
    SeqIF™ (sequential immunofluorescence) staining on COMET™ of Purified anti-Vimentin (clone O91D3, yellow) on formalin-fixed paraffin-embedded human pancreatic carcinoma at 0.33 µg/mL. Alexa Fluor™ Plus 647 Goat anti-Mouse IgG antibody (Lunaphore, Cat. No. DR647MS) was used as a secondary antibody. Nuclei were counterstained with DAPI (blue). Tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing.
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677801 25 µg 101 CHF
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677802 100 µg 265 CHF
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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.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

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

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.25 - 2.5 µg per mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 µg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 5.0 µg/mL is suggested. For immunohistochemistry on frozen tissue sections, a concentration range of 1.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

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 applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ 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. Rodríguez E, et al. 2022. Commun Biol. 5:41. PubMed
  2. Lin CY, et al. 2022. Int J Mol Sci. 23:. PubMed
RRID
AB_2565911 (BioLegend Cat. No. 677801)
AB_2565911 (BioLegend Cat. No. 677802)

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: 7    Revision Date: 11.17.2023

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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