Journal: Molecular Vision

Article Title: Structural basement membrane components and corresponding integrins in Schlemm's canal endothelia

doi:

Figure Lengend Snippet: Antibodies used for western blotting.

Article Snippet: Laminin α5 was labeled with rabbit anti-Laminin α5 IgG (Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:50 in PBS and incubated overnight at 4 0C, followed by incubation in Alexa-647 goat anti-rabbit IgG (Invitrogen, Austin, TX) diluted 1:150 for 3 h. Integrin α6 was labeled with rat anti-Integrin α6 (gift of Damsky Lab) diluted 1:50 in dPBS and incubated overnight at 4 °C, followed by incubation in Cy3 goat anti-rat IgG (Abcam, Cambridge, MA) diluted 1:100 in dPBS, for 3 h at RT.

Techniques: Western Blot

Journal: Molecular Vision

Article Title: Structural basement membrane components and corresponding integrins in Schlemm's canal endothelia

doi:

Figure Lengend Snippet: Laminins-332 and −511 in human conventional outflow pathway and cultured cells. A : Confocal immunofluorescence microscopy of endothelial laminins (LM) in radial sections through human conventional outflow tissues. Shown are representative images taken from one human donor eye of three that were examined. B : western blot analysis of endothelial laminin levels by different strains of cultured SC and TM cell monolayers isolated from different individual eye donors. Samples for laminin α5 blots were obtained after cell lysates were first immunoprecipitated with anti-laminin γ1 IgGs.

Article Snippet: Laminin α5 was labeled with rabbit anti-Laminin α5 IgG (Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:50 in PBS and incubated overnight at 4 0C, followed by incubation in Alexa-647 goat anti-rabbit IgG (Invitrogen, Austin, TX) diluted 1:150 for 3 h. Integrin α6 was labeled with rat anti-Integrin α6 (gift of Damsky Lab) diluted 1:50 in dPBS and incubated overnight at 4 °C, followed by incubation in Cy3 goat anti-rat IgG (Abcam, Cambridge, MA) diluted 1:100 in dPBS, for 3 h at RT.

Techniques: Cell Culture, Immunofluorescence, Microscopy, Western Blot, Isolation, Immunoprecipitation

Journal: Molecular Vision

Article Title: Structural basement membrane components and corresponding integrins in Schlemm's canal endothelia

doi:

Figure Lengend Snippet: En face confocal immunofluorescence microscopy of Schlemm’s canal inner wall. Shown is labeling of the inner wall of SC (viewed from luminal side) for integrin α6 ( A , green), laminin α5 ( B , red), CD31 ( C , blue). Panel D is a merged image of all three proteins plus nuclei (brown). Shown is representative image from a human donor eye of 6 total eyes that were examined.

Article Snippet: Laminin α5 was labeled with rabbit anti-Laminin α5 IgG (Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:50 in PBS and incubated overnight at 4 0C, followed by incubation in Alexa-647 goat anti-rabbit IgG (Invitrogen, Austin, TX) diluted 1:150 for 3 h. Integrin α6 was labeled with rat anti-Integrin α6 (gift of Damsky Lab) diluted 1:50 in dPBS and incubated overnight at 4 °C, followed by incubation in Cy3 goat anti-rat IgG (Abcam, Cambridge, MA) diluted 1:100 in dPBS, for 3 h at RT.

Techniques: Immunofluorescence, Microscopy, Labeling