Immunofluorescence Staining for Protein Localization

Immunofluorescence was performed as previously described [2 (link),3 (link),36 (link)] with antibodies recognizing the following antigens: Myosin Heavy Chain (DSHB; MF20), Sox9 (Novus; NBP1-85551), EGFP (Aves Lab; GFP-1020), and Islet 1 (R&D; AF1837). Secondary antibodies (Jackson Immunoresearch) included anti-chicken FITC (703-095-155), anti-mouse TRITC (715-025-150), anti-goat TRITC (705-025-147), and anti-rabbit Cy5 (711-605-152). Nuclei were visualized using DAPI (Invitrogen; Slowfade Gold Antifade Reagent with DAPI; catalogue#: S36938) and fluorescence visualized using a Zeiss AxioImager II microscope.

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Deepe R.N., Drummond J.R., Wolters R.A., Fitzgerald E.A., Tarolli H.G., Harvey A.B, & Wessels A. (2022). Sox9 Expression in the Second Heart Field; A Morphological Assessment of the Importance to Cardiac Development with Emphasis on Atrioventricular Septation. Journal of Cardiovascular Development and Disease, 9(11), 376.

Publication 2022

GFP-1020 AF1837 Slowfade Gold Antifade Reagent with DAPI AxioImager II microscope

Antibodies Antigens Aves Chicken Dapi Fitc Fluorescence Goat Gold Immunofluorescence Microscope Mouse Myosin heavy chain Novus Nuclei Rabbit Sox9 Tritc

Corresponding Organization : Medical University of South Carolina

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Variable analysis

independent variables

Antibodies recognizing the following antigens: Myosin Heavy Chain (DSHB; MF20), Sox9 (Novus; NBP1-85551), EGFP (Aves Lab; GFP-1020), and Islet 1 (R&D; AF1837)

dependent variables

Fluorescence visualization

control variables

Immunofluorescence protocol as previously described [2 (link),3 (link),36 (link)]
Secondary antibodies (Jackson Immunoresearch) included anti-chicken FITC (703-095-155), anti-mouse TRITC (715-025-150), anti-goat TRITC (705-025-147), and anti-rabbit Cy5 (711-605-152)
Nuclei visualized using DAPI (Invitrogen; Slowfade Gold Antifade Reagent with DAPI; catalogue#: S36938)
Fluorescence visualized using a Zeiss AxioImager II microscope

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