Mouse anti fibronectin

Manufactured by Merck Group

Sourced in Japan, United Kingdom, Sao Tome and Principe

Mouse anti-fibronectin is a laboratory reagent used to detect and quantify the presence of fibronectin, a glycoprotein found in the extracellular matrix. It is a monoclonal antibody raised against fibronectin and can be used in various immunoassay techniques to identify and measure fibronectin levels in biological samples.

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Close spelling variants of this product

Fibronectin (1886 citations) Anti-fibronectin (91 citations) Mouse anti-human fibronectin antibody (2 citations) Mouse monoclonal anti-fibronectin (2 citations)

7 protocols using mouse anti fibronectin

Immunolabeling of Neural Markers

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The following antibodies were used as primary antibodies for immunocytostaining and for immunohistochemistry: rabbit anti‐s100b (dilution = 1:1,000) (Dako, Ely, UK), rabbit anti‐p75 (1:500) (Cell Signaling Technology, Danvers, MA, USA), rabbit anti‐GAP43 (1:200) (Cell Signaling Technology), rabbit anti‐glial fibrillary acidic protein (GFAP) (1:200) (Abcam, Cambridge, UK), rabbit anti‐NG2 (1:1,000) (Millipore, Cambridge, UK), mouse anti‐myelin basic protein (MBP) (1:100) (Chemicon, Chandlers Ford, UK), goat anti‐protein zero (P0) (1:500) (Abcam), rabbit anti‐Tuj‐1 (1:100) (Chemicon), mouse anti‐neurofilament (NF) (1:1,000) (Sigma), rabbit anti‐PGP9.5 (1:500) (Abcam), mouse anti‐fibronectin (Millipore), mouse anti‐Nanog (1:200) (ReproCELL, Yokohama, Japan), mouse anti‐MAP‐2 (1:200) (Abcam), and rabbit anti‐NeuN (1:500) (Abcam) antibodies. As secondary antibodies, Alexa Fluor 546‐conjugated anti‐rabbit and anti‐mouse antibodies and Alexa Fluor 670‐conjugated anti‐rabbit and anti‐mouse antibodies (1:1,000) (Life technologies, Carlsbad, CA, USA) were used.

Sowa Y., Kishida T., Tomita K., Yamamoto K., Numajiri T, & Mazda O. (2017). Direct Conversion of Human Fibroblasts into Schwann Cells that Facilitate Regeneration of Injured Peripheral Nerve In Vivo. Stem Cells Translational Medicine, 6(4), 1207-1216.

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Immunofluorescence Analysis of Embryonic Gonads

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Control and DMRT1 over-expressing tissues were fixed for 15 minutes in 4% PFA/PBS at room temperature, prior to processing for tissue section immunofluorescence, as described previously (Smith et al. 2003 (link)). Controls comprised either uninfected stage-matched embryos, or those infected with RCASBP.EGFP. At least three embryos per time point and/or treatment were examined. Since the embryonic gonad is elongated and sausage-shaped, serial longitudinal sections were cut, which allowed a more thorough analysis of over-expression along the length of the gonad, compared with transverse sections. Briefly, 10 µm sections were cut on a cryostat, permeablised in PBS+1% Triton X-100 and blocked in PBS+2% BSA for 1 hour. Primary antibodies were added overnight at 4°C and were either raised in-house (rabbit anti-chicken aromatase at 1:5000, rabbit anti-chicken DMRT1 at 1:5000; rabbit anti-HEMGN at 1:300), or were obtained commercially (Charles River Services; rabbit anti-p27 (1:1000), Santa Cruz; goat anti-human AMH polyclonal (1:1000), Serotec; mouse antifibronectin (1:500), Millipore; goat anti-GFP (1:500), Millipore; Rabbit anti-human SOX9 (1:6000)). Alexa-fluor secondary antibodies were used (donkey or goat anti-rabbit, mouse or goat 488 or 594; Molecular Probes). Sections were counterstained with DAPI and images collected on a Zeiss microscope equipped with fluorescence optics.

Lambeth L., Raymond C.S., Roeszler K.N., Kuroiwa A., Nakata T., Zarkower D, & Smith C.A. (2014). Over-expression of DMRT1 induces the male pathway in embryonic chicken gonads. Developmental biology, 389(2), 160-172.

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Immunoblotting for Protein Detection

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Standard immunoblotting procedures51 (link) were used to detect LMP1 (with mouse CS1-4 at 1:5), activin A (with mouse anti-inhibin βA at 1:50 [Serotec, Oxfordshire, UK]), β-actin (with mouse anti-β-actin at 1:20,000 [Sigma]), fibronectin (with mouse anti-fibronectin at 1:100 [Sigma]), phosphorylated Smad2 Ser465/Ser467 (with rabbit anti-pSmad2 at 1:250 [Cell Signaling Technology, Hitchin, UK]), Smad2 (with rabbit anti-Smad2 at 1:250 [Cell Signaling Technology]), Smad4 (with rabbit anti-Smad4 at 1:250 [Cell Signaling Technology]), phosphorylated JNK/SAPK (with rabbit anti-pJNK/SAPK at 1:500 [Cell Signaling Technology]), JNK/SAPK (with rabbit anti-JNK/SAPK at 1:500 [Cell Signaling Technology]).

Morris M.A., Dawson C.W., Laverick L., Davis A.M., Dudman J.P., Raveenthiraraj S., Ahmad Z., Yap L.F, & Young L.S. (2016). The Epstein-Barr virus encoded LMP1 oncoprotein modulates cell adhesion via regulation of activin A/TGFβ and β1 integrin signalling. Scientific Reports, 6, 19533.

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Multicolor Immunofluorescence Staining of Cardiomyocytes

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Coverslips with cardiomyocytes and fibroblasts were fixed and permeabilized with 4% formaldehyde and 0.1% Triton-X 100 in PBS for 15 min and then washed 3 times with PBS. Coverslips were then stained with dilutions of 1:200 DAPI, 1:75 phalloidin conjugated to Alexa Fluor 488, and 1:200 primary antibodies, mouse anti-sarcomeric-α-actinin (Sigma A7811) or mouse anti-fibronectin (Sigma F3648). Samples were incubated with these stains for 60 min and then washed 3 times with PBS. Samples were then incubated in 1:100 dilution of goat anti-mouse secondary antibodies conjugated to Alexa Fluor 555 for 60 min and the washed 3 times in PBS. Finally, coverslips were mounted for imaging onto glass slides with ProLong preservative. All samples were imaged using confocal laser scanning microscopy (LSM 700, Carl Zeiss Microscopy, LLC).

Batalov I., Jallerat Q., Kim S., Bliley J, & Feinberg A.W. (2021). Engineering aligned human cardiac muscle using developmentally inspired fibronectin micropatterns. Scientific Reports, 11, 11502.

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Immunophenotyping of GS-1 Cell Line

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Immunophenotyping of the GS-1 cell line was carried out at the 30th passage following the method described previously [10 (link)] with some modifications.
Briefly, the cells were grown on coverslips in six-well plates for 24 hr and then fixed with 4% paraformaldehyde at 4°C for 1 hr and permeabilized with 0.2% Triton-X 100 at room temperature
for 15 min. After blocking with 2% bovine serum albumin (BSA) in PBS at room temperature for 2 hr, the cells were then incubated with four different primary antibodies of
mouse-anti-cytokeratin, mouse-anti-vimentin, mouse-anti-fibronectin and rabbit-anti-desmin (Sigma-Aldrich Co., St. Louis, MO, U.S.A.) at a dilution of 1:200 at room temperature for 2 hr to
characterize the cell line. Subsequently, the cells were rinsed three times with PBS and were incubated with the appropriate secondary antibodies of goat-anti-mouse FITC-conjugated or
goat-anti-rabbit FITC-conjugated (Sigma-Aldrich) at a dilution of 1:320 in PBS containing 1% BSA at room temperature for 1 hr. In control coverslips, only PBS with 1% BSA was used in place
of the primary antibodies. Diaminophenylindole at a final concentration of 1 µg/ml was used to stain DNA in nuclei. All samples were observed with a Carl
Zeiss fluorescence microscope.

HUANG S.M., KUO S.T., KUO H.C, & CHANG S.K. (2018). Assessment of fish iridoviruses using a novel cell line GS-1, derived from the spleen of orange-spotted grouper Epinephelus coioides (Hamilton) and susceptible to ranavirus and megalocytivirus. The Journal of Veterinary Medical Science, 80(11), 1766-1774.

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Immunohistochemical Analysis of Embryos

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Immunohistochemistry was performed according to standard protocols. Fibronectin staining was performed according to a protocol provided by Scott Holley. Stained embryos were mounted in 1% low-melt agarose and imaged on an LSM800 confocal microscope or Spinning Disk confocal microscope (Zeiss). Primary antibodies used were chicken anti-GFP (1:200, Aves Labs), rabbit anti-phospho-Histone H3 (1:2000, Millipore), rabbit anti-cleaved Caspase-3 (1:200, Cell Signaling), mouse anti-Fibronectin (1:100, Sigma). Secondary antibodies (Life Technologies) were used at 1:300. Positive cells were counted using FIJI/Image J.

Collins M.M., Maischein H.M., Dufourcq P., Charpentier M., Blader P, & Stainier D.Y. (2018). Pitx2c orchestrates embryonic axis extension via mesendodermal cell migration. eLife, 7, e34880.

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Fibroblast Extracellular Matrix Characterization

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Human dermal fibroblasts cultured on glass coverslips were fixed and permeabilized in 4% PFA with 0.1% Triton X and blocked with 1% BSA, before staining with mouse anti‐vimentin (Sigma‐Aldrich), mouse anti‐fibronectin (Sigma‐Aldrich) and rabbit anti‐collagen type I (Abcam) for 1 h at 37°C. After washing the primary antibody, the cells were incubated with secondary antibodies conjugated with: rabbit Alexa 488 or mouse Alexa 568 (Thermo Scientific). The coverslips were mounted with Fluoroshield with DAPI (Thermo Scientific) and visualized using a Zeiss Observer D1 microscope.

Tutuianu R., Rosca A., Albu Kaya M.G., Pruna V., Neagu T.P., Lascar I., Simionescu M, & Titorencu I. (2020). Mesenchymal stromal cell‐derived factors promote the colonization of collagen 3D scaffolds with human skin cells. Journal of Cellular and Molecular Medicine, 24(17), 9692-9704.

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