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Fibronectin_A647_TDS_Flow.png

Flow cytometric analysis of fibronectin in human mesenchymal stem cells (MSC).  MSC (Lonza), passage 6, were dissociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Anti-Fibronectin monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD LSRFortessa™ II flow cytometry system. BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) is also suitable for permeabilization.

Fibronectin_A647_TDS_Flow.png

Flow cytometric analysis of fibronectin in human mesenchymal stem cells (MSC).  MSC (Lonza), passage 6, were dissociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Anti-Fibronectin monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD LSRFortessa™ II flow cytometry system. BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) is also suitable for permeabilization.

Fibronectin_A647_TDS_D12_Merged.png

Immunofluorescent analysis of fibronectin in human 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® 647 Mouse Anti-Fibronectin monoclonal antibody (pseudo-colored red) at 1.2 µg/ml. Cell nuclei were stained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™Software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.

Fibronectin_A647_TDS_D12_Merged.png

Immunofluorescent analysis of fibronectin in human 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® 647 Mouse Anti-Fibronectin monoclonal antibody (pseudo-colored red) at 1.2 µg/ml. Cell nuclei were stained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™Software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.

Fibronectin_A647_TDS_Flow.png Fibronectin_A647_TDS_Flow.png Fibronectin_A647_TDS_D12_Merged.png Fibronectin_A647_TDS_D12_Merged.png Red-Cap-Red-Label.jpg

Flow cytometric analysis of fibronectin in human mesenchymal stem cells (MSC).  MSC (Lonza), passage 6, were dissociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Anti-Fibronectin monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD LSRFortessa™ II flow cytometry system. BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) is also suitable for permeabilization.

Flow cytometric analysis of fibronectin in human mesenchymal stem cells (MSC).  MSC (Lonza), passage 6, were dissociated and fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ isotype control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Anti-Fibronectin monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MSC. Flow cytometry was performed on a BD LSRFortessa™ II flow cytometry system. BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) is also suitable for permeabilization.

Immunofluorescent analysis of fibronectin in human 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® 647 Mouse Anti-Fibronectin monoclonal antibody (pseudo-colored red) at 1.2 µg/ml. Cell nuclei were stained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™Software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.

Immunofluorescent analysis of fibronectin in human 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® 647 Mouse Anti-Fibronectin monoclonal antibody (pseudo-colored red) at 1.2 µg/ml. Cell nuclei were stained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™Software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.

Product Details
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BD Pharmingen™
FN; LETS
Human (QC Testing), Mouse, Rat, Dog, Chicken, Cow (Reactivity Confirmed in Development)
Mouse IgG1, κ
Human Fibronectin
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_2738004
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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. 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.
  7. Triton is a trademark of the Dow Chemical Company.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. 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.
  10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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563098 Rev. 1
Antibody Details
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10/Fibronectin

The 240-kDa dimeric fibronectin protein exists in two forms: a soluble protomer in body fluids and an insoluble multimer in the extracellular matrix. The latter is the primary functional form and creates a substrate for cell migration, a role which makes fibronectin vital to embryogenesis and wound response. Fibronectin mediates cytoskeletal organization, cell attachment, and cellular signaling through interactions with cellular receptors. Although various isoforms of fibronectin are derived by alternative splicing, they share a common N-terminus which is a critical region for cell surface binding in an initial step of multimer assembly. Further polymerization steps are regulated by fibronectin/integrin interactions and result in generation of the complex fibrils that constitute the fibronectin matrix.

563098 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Alexa Fluor™ 647
653 nm
669 nm
563098 Rev.1
Citations & References
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View product citations for antibody "563098" on CiteAb

Development References (5)

  1. Chen H, Mosher DF. Formation of sodium dodecyl sulfate-stable fibronectin multimers. Failure to detect products of thiol-disulfide exchange in cyanogen bromide or limited acid digests of stabilized matrix fibronectin. J Biol Chem. 1996; 271(15):9084-9089. (Biology). View Reference
  2. Danen EH, Sonneveld P, Brakebusch C, Fassler R, Sonnenberg A. The fibronectin-binding integrins alpha5beta1 and alphavbeta3 differentially modulate RhoA-GTP loading, organization of cell matrix adhesions, and fibronectin fibrillogenesis. J Cell Biol. 2002; 159(6):1071-1086. (Clone-specific: Immunofluorescence). View Reference
  3. Rhee CS, Sen M, Lu D, et al. Wnt and frizzled receptors as potential targets for immunotherapy in head and neck squamous cell carcinomas. Oncogene. 2002; 21(42):6598-6605. (Clone-specific: Western blot). View Reference
  4. Sechler JL, Takada Y, Schwarzbauer JE. Altered rate of fibronectin matrix assembly by deletion of the first type III repeats. J Cell Biol. 1996; 134(2):573-583. (Biology). View Reference
  5. Zuk A, Bonventre JV, Brown D, Matlin KS. Polarity, integrin, and extracellular matrix dynamics in the postischemic rat kidney. Am J Physiol. 1998; 275(3):C711-C731. (Clone-specific: Immunohistochemistry). View Reference
View All (5) View Less
563098 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.

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.