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anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free)

AG-20B-6001PF-C100 100 µg INQ
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Additional Information

Product Data
Synonyms FN; Cold-insoluble Globulin; CIG; Fibronectin (EIIIA)
Properties
Clone IST-9
Isotype Mouse IgG1
Source/Host Purified from concentrated hybridoma tissue culture supernatant.
Immunogen/Antigen Recombinant Extra Domain A (EDA) region of human cellular fibronectin.
Application ELISA
Functional Application: Inhibits binding of Fibronectin EDA domain to α5-β1, α4-β1 and α9-β1 integrins. Inhibits cell adhesion mediated by α5-β1, α4-β1 and α9-β1 integrins. Blocks the differentiation of fibroblasts into myo-fibroblasts and the TGF-β1-triggered enhancement of α-smooth muscle actin and collagen type I.
Immunocytochemistry: 1:200
Immunohistochemistry: (frozen sections: 1-2μg/ml and paraffin sections: 2-10μg/ml)
Western Blot: 1:1000
Crossreactivity Chicken, Cow, Dog, Human, Monkey, Mouse, Pig, Rat
Specificity Recognizes an epitope located in the EDA sequence of human cellular fibronectin (FN). Does not detect extracellular FN (plasma FN). Cross-reacts with mouse, rat, monkey, pig, dog, cow and chicken fibronectin EDA (according to literature).
Purity ≥95% (SDS-PAGE)
Purity Detail Epitope affinity purifed on recombinant fragment of fibronectin EDA.
Endotoxin Content <0.001EU/μg purified protein (LAL test; Lonza).
Concentration Lot dependent
Formulation Liquid. In 20mM sodium phosphate buffer, pH7.6 containing 150mM sodium chloride.
Product Type Monoclonal Antibody
Shipping and Handling
Shipping BLUE ICE
Short Term Storage +4°C
Long Term Storage -20°C
Handling Advice After opening, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.
Use/Stability Stable for at least 1 year after receipt when stored at -20°C.
Documents
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Product Description

Fibronectin is a major component of the extracellular matrix that binds to integrins, cell surfaces and various compounds including collagen, fibrin, heparin, DNA and actin. It occurs in two main forms: plasma and cellular fibronectin. EDA (Extra Domain A)-containing cellular fibronectin (EDA-FN) plays a major role in cell adhesion, growth, migration and differentiation and it is important for processes such as wound healing and embryonic development. It is associated with fibroblast differentiation by increasing α-SMA expression, collagen deposition, cell contractility and focal adhesion kinase (FAK) activation. EDA-FN promotes cell spread by interacting with the integrins α5-β1, α4-β1 and α9-β1 and has been shown to promote angiogenesis, lymphangiogenesis and metastasis of colorectal tumors. In transformed or tumor-derived cells the EDA segment is about 10-times higher than in fibronectin from normal fibroblasts and it may therefore be a significant marker for malignancy.

Product References

  1. Monoclonal antibodies in the analysis of fibronectin isoforms generated by alternative splicing of mRNA precursors in normal and transformed human cells: L. Borsi, et al.; J. Cell Biol. 104, 595 (1987)
  2. Localization of the cellular-fibronectin-specific epitope recognized by the monoclonal antibody IST-9 using fusion proteins expressed in E. coli: B. Carnemolla, et al.; FEBS Lett. 215, 269 (1987)
  3. Molecular and immunologic differences in canine fibronectins from articular cartilage and plasma: N. Burton-Wurster & G. Lust; Arch. Biochem. Biophys. 269, 32 (1989)
  4. Coordinate oncodevelopmental modulation of alternative splicing of fibronectin pre-messenger RNA at ED-A, ED-B, and CS1 regions in human liver tumors: F. Oyama, et al.; Cancer Res. 53, 2005 (1993)
  5. Endogenous fibronectin of blood polymorphonuclear leukocytes: immunochemical characterization and subcellular localization: R. Salcedo, et al.; Exp. Cell Res. 233, 25 (1997)
  6. The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-beta1: G. Serini, et al.; J. Cell Biol. 142, 873 (1998)
  7. Identification of two amino acids within the EIIIA (ED-A) segment of fibronectin constituting the epitope for two function-blocking monoclonal antibodies: Y.F. Liao, et al.; J. Biol. Chem. 274, 17876 (1999)
  8. Engagement of alpha4beta7 integrins by monoclonal antibodies or ligands enhances survival of human eosinophils in vitro: J. Meerschaert, et al.; J. Immunol. 163, 6217 (1999)
  9. Segmental antigen challenge increases fibronectin in bronchoalveolar lavage fluid: J. Meerschaert, et al.; Am. J. Respir. Crit. Care Med. 159, 619 (1999)
  10. Deletion of the alternatively spliced fibronectin EIIIA domain in mice reduces atherosclerosis: M.H. Tan, et al.; Blood 104, 11 (2004)
  11. Fibronectin-alpha4beta1 integrin interactions regulate metalloproteinase-9 expression in steatotic liver ischemia and reperfusion injury: C. Moore, et al.; Am. J. Pathol. 170, 567 (2007)
  12. Identification of the peptide sequences within the EIIIA (EDA) segment of fibronectin that mediate integrin alpha9beta1-dependent cellular activities: A.V. Shinde, et al.; J. Biol. Chem. 283, 2858 (2008)
  13. Fibronectin extra domain A (EDA) sustains CD133+/CD44+ subpopulation of colorectal cancer cells: J. Ou, et al.; Stem Cell Res. 11, 820 (2013)
  14. Irigenin, a novel lead from Western Himalayan chemiome inhibits Fibronectin-Extra Domain A induced metastasis in lung cancer cells: A. Amin, et al.; Sci. Rep. 6, 37151 (2016)
  15. Detection of soluble ED-A+ fibronectin and evaluation as novel serum biomarker for cardiac tissue remodeling: B. Ziffels, et al.; Dis. Mark. 2016, ID3695454 (2016)
Immunohistochemical analysis on paraffin-embedded sections of human melanoma SK-MEL-28 using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF).
Immunohistochemical analysis on paraffin-embedded sections of human melanoma SK-MEL-28 using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF).
Immunocytochemical analysis of human esophageal epithelial cells using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF). An anti-mouse IgG secondary antibody was used. Nuclei were counterstained with propidium iodide.
Immunocytochemical analysis of human esophageal epithelial cells using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF). An anti-mouse IgG secondary antibody was used. Nuclei were counterstained with propidium iodide.
Western blot analysis of human dermal fibroblast whole cell lysate, untreated (left) and treated with TGF-&beta;1 (right) using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF). Lysates/proteins at 10 µg per lane.  A goat anti-mouse HRP-conjugated monoclonal secondary antibody was used.
Western blot analysis of human dermal fibroblast whole cell lysate, untreated (left) and treated with TGF-β1 (right) using anti-Fibronectin (EDA), mAb (blocking) (IST-9) (preservative free) (AG-20B-6001PF). Lysates/proteins at 10 µg per lane. A goat anti-mouse HRP-conjugated monoclonal secondary antibody was used.
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