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Alexa Fluor® 647 anti-Vimentin Antibody

Pricing & Availability
Clone
W16220A (See other available formats)
Regulatory Status
RUO
Other Names
Vim, CTRCT30, HEL 113, Epididymis Luminal Protein 113
Isotype
Rat IgG2a, κ
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Product Citations
publications
W16220A_A647_Vimentin_Antibody_102320
NIH/3T3 cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 6 minutes, and blocked with 5% FBS for 30 minutes. Cells were then intracellularly stained with 2.0 µg/mL (1:250 dilution) Alexa Fluor® 647 rat IgG2a, κ isotype ctrl antibody (Cat. No. 400526) (panel A) or Alexa Fluor® 647 anti-vimentin antibody (clone W16220A) (panel B) overnight at 4°C. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • W16220A_A647_Vimentin_Antibody_102320
    NIH/3T3 cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 6 minutes, and blocked with 5% FBS for 30 minutes. Cells were then intracellularly stained with 2.0 µg/mL (1:250 dilution) Alexa Fluor® 647 rat IgG2a, κ isotype ctrl antibody (Cat. No. 400526) (panel A) or Alexa Fluor® 647 anti-vimentin antibody (clone W16220A) (panel B) overnight at 4°C. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • W16220A_A647_Vimentin_Antibody_2_120621
    EL-4 cells (positive target) were fixed with fixation buffer (Cat. No. 42080) and permeabilized with True-Phos™ Perm Buffer (Cat. No. 425401), then intracellularly stained with 0.125 µg/test of Alexa Fluor® 647 anti-Vimentin antibody (clone W16220A) (filled histogram) or Alexa Fluor® 647 rat IgG2a, κ isotype ctrl (open histogram).
  • W16220A_A647_Vimentin_Antibody_3_120621
    A20 cells (negative target) were fixed with fixation buffer Cat. No. 420801 and permeabilized with True-Phos™ Perm Buffer (Cat. No. 425401), then intracellularly stained with 0.125 µg/test of Alexa Fluor® 647 anti-Vimentin antibody (clone W16220A) (filled histogram) or Alexa Fluor® 647 rat IgG2a, κ isotype ctrl (open histogram).
Compare all formats See Alexa Fluor® 647 spectral data
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699307 25 µg £109
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699308 100 µg £254
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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
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial mouse Vimentin (224-446 a.a.) recombinant protein expressed 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® 647 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 - Verified
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


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

View full statement regarding label licenses
Excitation Laser
Red Laser (633 nm)
Application Notes

This antibody does not cross-react with human (in-house tested).

May be weakly reactive with human Vimentin by WB.

Additional Product Notes

This product has been verified for IHC-P (Immunohistochemistry - formalin-fixed paraffin-embedded tissues) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

RRID
AB_2888890 (BioLegend Cat. No. 699307)
AB_2888890 (BioLegend Cat. No. 699308)

Antigen Details

Structure
466 amino acids with a predicted molecular weight of 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.
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
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
22352 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: 3    Revision Date: 01/24/2024

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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