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Visualization of Focal Adhesions in 3T3 Cells Using Anti-Vinculin and Phalloidin Staining

This study focuses on the visualization of focal adhesions in 3T3 cells by employing immunohistochemical techniques. We utilized anti-vinculin monoclonal antibody as a primary antibody, followed by a goat anti-mouse secondary antibody coupled with FITC, to specifically detect vinculin protein. Additionally, phalloidin conjugated to TRITC was used to visualize actin filaments, allowing for an in-depth analysis of cellular adhesion dynamics. Various experimental groups were prepared to compare control and experimental conditions, highlighting the presence of focal adhesions effectively.

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Visualization of Focal Adhesions in 3T3 Cells Using Anti-Vinculin and Phalloidin Staining

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  1. BIOTECH 2 IMMUNOHISTOCHEM LAB SPRING 12 2/28/2012

  2. 3T3 CELLS To visualize focal adhesions having vinculin protien • PRIMARY AB: anti-vinculinmAb • Secondary Ab:goat anti mouse coupled to FITC To visualize actin as phallodin binds actin Phallodin coupled to TRITC

  3. Phallodin coupled to TRITC [To visualize actin as phallodin binds actin]

  4. PRIMARY AB: anti-vinculinmAbSecondary Ab:goat anti mouse To visualize focal adhesions having coupled to FITC vinculin protein

  5. Example of what you should have seen

  6. Grp 1 : control primary +control secondary+DAPI DAPI DAPI FITC FITC TRITC TRITC Grp 2 : control primary + phallodin TRITC

  7. Grp 3 : CONTROL PRIMARY+SECNDRY AB-FITC+DAPI DAPI FITC TRITC DAPI FITC TRITC Grp 4 : anti-vinculin+contl secondary+ DAPI

  8. Grp 5: anti vinculin + sec ab-FITC DAPI DAPI FITC FITC TRITC TRITC Grp 6 : anti vinculin+ phallodin+sec ab-FITC +DAPI

  9. Grp 7 : contrl primary +phallodin-TRITC + DAPI DAPI DAPI FITC FITC TRITC TRITC Grp 8 :anti-vinculin +phallodin+ secondary-ab-FITC

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