ECM-producing HPSC cultures were fixed, permeabilized and processed as published (30 ). Samples were blocked using Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE) containing 1% (v/v) donkey serum then incubated with either rabbit polyclonal anti-human fibronectin (1∶200) or 1A4 mouse anti-αSMA (1∶300), both from Sigma-Aldrich and diluted in blocking buffer. Fluorophore cross-linked secondary antibodies (Jackson ImmunoResearch Laboratories Inc.) were incubated at 1∶100 dilution. Samples were mounted using Prolong Gold anti-fading reagent from Life Technologies (Carlsbad, CA) and scanned using a confocal spinning disk Ultraview (Perkin-Elmer Life Sciences, Boston, MA) microscope. Images were acquired with a 60× 1.45 PlanApo TIRF oil immersion objective, under identical exposure conditions using Volocity 6.3.0 (Perkin-Elmer Life Sciences).
Rabbit polyclonal anti human fibronectin
Manufactured by Merck Group
Rabbit polyclonal anti-human fibronectin is a laboratory reagent used for the detection and quantification of human fibronectin. It is produced by immunizing rabbits with purified human fibronectin, and the resulting polyclonal antibodies are specific for this protein. This product is intended for research use only.
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Lab products found in correlation
2 protocols using rabbit polyclonal anti human fibronectin
1
Quantifying ECM Protein Expression in HPSCs
2
Quantifying ECM Protein Expression in HPSCs
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ECM-producing HPSC cultures were fixed, permeabilized and processed as published (30 ). Samples were blocked using Odyssey® Blocking Buffer (LI-COR Biosciences, Lincoln, NE) containing 1% (v/v) donkey serum then incubated with either rabbit polyclonal anti-human fibronectin (1∶200) or 1A4 mouse anti-αSMA (1∶300), both from Sigma-Aldrich and diluted in blocking buffer. Fluorophore cross-linked secondary antibodies (Jackson ImmunoResearch Laboratories Inc.) were incubated at 1∶100 dilution. Samples were mounted using Prolong Gold anti-fading reagent from Life Technologies (Carlsbad, CA) and scanned using a confocal spinning disk Ultraview (Perkin-Elmer Life Sciences, Boston, MA) microscope. Images were acquired with a 60× 1.45 PlanApo TIRF oil immersion objective, under identical exposure conditions using Volocity 6.3.0 (Perkin-Elmer Life Sciences).
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