Western blot analysis of Fibronectin. A-431 Cell Lysate (Cat. No. 611447) was blotted with Purified Mouse Anti-Fibronectin monoclonal antibody at dilutions of 1:5000 (Lane 1), 1:10,000 (Lane 2), and 1:20,000 (Lane 3).
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 Purified Mouse Anti-Fibronectin monoclonal antibody (Cat. No.610077, pseudo-colored green) at 2.5 µg/ml. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies), and 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.
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). Cells were stained with Purified Mouse IgG1, κ isotype control (dashed line, Cat. No. 349040) or Purified Mouse Anti-Fibronectin monoclonal antibody (solid line, Cat. No. 610077) at matched concentrations. The secondary antibody was FITC Goat Anti-Mouse Ig (Cat No. 554001). Histograms were derived from gated events based on light scattering characteristics of MSCs. 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.
Western blot analysis of Fibronectin. A-431 Cell Lysate (Cat. No. 611447) was blotted with Purified Mouse Anti-Fibronectin monoclonal antibody at dilutions of 1:5000 (Lane 1), 1:10,000 (Lane 2), and 1:20,000 (Lane 3).
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 Purified Mouse Anti-Fibronectin monoclonal antibody (Cat. No.610077, pseudo-colored green) at 2.5 µg/ml. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies), and 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.
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). Cells were stained with Purified Mouse IgG1, κ isotype control (dashed line, Cat. No. 349040) or Purified Mouse Anti-Fibronectin monoclonal antibody (solid line, Cat. No. 610077) at matched concentrations. The secondary antibody was FITC Goat Anti-Mouse Ig (Cat No. 554001). Histograms were derived from gated events based on light scattering characteristics of MSCs. 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.
Product Details
BD Transduction Laboratories™
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), Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot (Tested During Development)
240 kDa
250 µg/ml
AB_397486
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Triton is a trademark of the Dow Chemical Company.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
Purified Mouse IgG1, κ Isotype Control RUO (GMP)
Size
100 Tests
Cat No.
349040
FITC Goat Anti-Mouse Ig RUO
Size
0.5 mg
Cat No.
554001
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HRP Goat Anti-Mouse Ig RUO
Size
1 mL
Cat No.
554002
610077 Rev. 3
Antibody Details
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.
610077 Rev. 3
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
610077 Rev.3
Citations & References
Development References (5)
-
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
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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
-
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
-
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
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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
610077 Rev. 3
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.
