PerCP/Cyanine5.5 anti-Vimentin

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_PerCPCyanine55_Vimentin_Antibody_1_050124 — Daudi cells (negative control, open histogram) or Jurkat cells (positive control, filled histogram) were fixed and permeabilized using the True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with PerCP/Cyanine5.5 anti-Vimentin (clone O91D3) or PerCP/Cyanine5.5 Mouse IgG2a, κ Isotype Control (open histogram, dashed line) (representative histogram for both cell lines) (Cat. No. 400257).

Compare all formats See PerCP/Cyanine5.5 spectral data

Cat #	Size	Price	Save
677813	25 tests	¥38,720
677814	100 tests	¥86,460

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 leukocytes^1,2. 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

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 and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with PerCP/Cyanine5.5 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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: ICFC - Quality tested
SB - Community Verified
Recommended Usage: Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Phos™ Perm Buffer in Cell Suspensions Protocol. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

* PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.
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 intracellular flow cytometry (ICFC), fixation/permeabilization using True-Phos™ Perm Buffer (Cat. No. 425401) is recommended.

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.
RRID: AB_3106228 (BioLegend Cat. No. 677813)
AB_3106228 (BioLegend Cat. No. 677814)

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

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Related FAQs

How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?: PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.
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.

Other Formats

View All Vimentin Reagents Request Custom Conjugation

Description	Clone	Applications
Alexa Fluor® 594 anti-Vimentin	O91D3	ICC,IHC-P,SB
Purified anti-Vimentin	O91D3	WB,ICC,ICFC,IHC-P,IHC-F,SB
Direct-Blot™ HRP anti-Vimentin	O91D3	WB
Alexa Fluor® 647 anti-Vimentin	O91D3	ICC,IHC-P,SB
Alexa Fluor® 488 anti-Vimentin	O91D3	ICC,ICFC,IHC-P,SB
TotalSeq™-Bn1302 anti-Vimentin	O91D3	SB
PerCP/Cyanine5.5 anti-Vimentin	O91D3	ICFC,SB

Certificate of Analysis

Go To Top Version: 2 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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