BD Pharmingen™ Alexa Fluor® 488 Mouse Anti-Human Vimentin
Clone RV202 (RUO)
Flow cytometric analysis of vimentin in human embryonic stem (ES) cells differentiated to a neural fate. H9 human ES cells (WiCell, Madison, WI) passage 48 were differentiated with media containing Noggin (R&D Systems) for 17 days (Yuan SH, Martin J, Elia J, et al, 2011), fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050), and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557721) or Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, solid line). Histograms were derived from gated events based on light scattering characteristics of the H9-derived ectoderm. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of vimentin in mesenchymal stem cells (MSC). MSC (Lonza) were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050), and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (dashed line, Cat. 557721) or Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, solid line). Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Immunofluorescent analysis of vimentin in mesenchymal stem cells (MSC). MSC (Lonza), passage 6, were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 and stained with Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, pseudo-colored green). Counter-staining of cell nuclei was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
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Flow cytometric analysis of vimentin in human embryonic stem (ES) cells differentiated to a neural fate. H9 human ES cells (WiCell, Madison, WI) passage 48 were differentiated with media containing Noggin (R&D Systems) for 17 days (Yuan SH, Martin J, Elia J, et al, 2011), fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050), and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557721) or Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, solid line). Histograms were derived from gated events based on light scattering characteristics of the H9-derived ectoderm. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of vimentin in mesenchymal stem cells (MSC). MSC (Lonza) were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050), and stained with matching concentrations of either Alexa Fluor® 488 Mouse IgG1, κ isotype control (dashed line, Cat. 557721) or Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, solid line). Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Immunofluorescent analysis of vimentin in mesenchymal stem cells (MSC). MSC (Lonza), passage 6, were fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 and stained with Alexa Fluor® 488 Mouse Anti-Vimentin monoclonal antibody (Cat. No. 562338, pseudo-colored green). Counter-staining of cell nuclei was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
Product Details
BD Pharmingen™
Human (QC Testing)
Mouse IgG1
Purified Cow Lens Vimentin
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_10896994
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Triton is a trademark of the Dow Chemical Company.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor® 488 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557721
562338 Rev. 1
Antibody Details
RV202
Intermediate filaments (IF) are a subset of cytoskeletal proteins which function to give overall structural integrity to the plasma membrane as well to organize cells into specific tissues. IF proteins can be divided into six major types based upon the similarity in sequence. Vimentin belongs to the type III category of IF proteins which are expressed in leukocytes, blood vessel endothelial cells, some epithelial cells, and mesenchymal cells. Vimentin is also expressed together with several other IF proteins during the early stages of development. Vimentin is exchanged for the tissue-specific intermediate filament type as differentiation proceeds. Vimentin migrates in SDS/PAGE as a ~57 kDa protein.
562338 Rev. 1
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
562338 Rev.1
Citations & References
View product citations for antibody "562338" on CiteAb
Development References (6)
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Broers JL, Carney DN, Klein Rot M, et al . Intermediate filament proteins in classic and variant types of small cell lung carcinoma cell lines: a biochemical and immunochemical analysis using a panel of monoclonal and polyclonal antibodies. J Cell Sci. 1986; 83:37-60. (Clone-specific: Immunofluorescence, Western blot). View Reference
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Lodish HF. Molecular cell biology, 4th ed.. New York: W.H. Freeman; 2000:795-847.
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Pieper FR, Schaart G, Krimpenfort PJ, et al. Transgenic expression of the muscle-specific intermediate filament protein desmin in nonmuscle cells. J Cell Biol. 1989; 108(3):1009-1024. (Clone-specific: Electron microscopy, Immunofluorescence, Immunohistochemistry, Western blot). View Reference
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Raats JM, Pieper FR, Vree Egberts WT, Verrijp KN, Ramaekers FC, Bloemendal H. Assembly of amino-terminally deleted desmin in vimentin-free cells. J Cell Biol. 1990; 111(5):1971-1985. (Clone-specific: Immunohistochemistry). View Reference
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Viebahn C, Lane EB, Ramaekers FC. Keratin and vimentin expression in early organogenesis of the rabbit embryo. Cell Tissue Res. 1988; 253(3):553-562. (Clone-specific: Immunofluorescence). View Reference
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Yuan SH, Martin J, Elia J, et al. Cell-Surface Marker Signatures for the Isolation of Neural Stem Cells, Glia and Neurons Derived from Human Pluripotent Stem Cells. PLoS ONE. 6(3)(Methodology: Cell culture). View Reference
562338 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.