Hematoxylin & Eosin staining was performed as described (Wang et al., 2008 (link)). For flat mount staining of the choroidal vasculature, pigment bleaching in RPE cells and choroidal melanocytes was performed as described (Kim and Assawachananont, 2016 ). Briefly, pigmented eyes were fixed in 4% paraformaldehyde PBS for 30 min to 2 h. After washing with PBS three times, the cornea and lens were removed before 3% hydrogen peroxide bleaching at 55 °C for 30 min to 2 h depending on the stage, or at 4 °C for several days. Immunofluorescence staining was performed using standard procedures. Primary antibodies used include: Beta-galactosidase (LacZ) from chicken (1:300, Abcam), Endomucin from rat (1:200, Santa Cruz Biotechnology), PECAM-1 from rat (1:50, BD Pharmingen), ICAM-2 from rat (1: 200, BD Pharmingen). These three EC markers, Endomucin, PECAM-1 and ICAM-2, were used interchangeably, but some antibodies worked better than others due to different sample processing. Secondary antibodies include: goat antichicken (or rat) AlexaFluor 488 (1:500, Invitrogen) and goat anti-rat AlexaFluor 594 (1:500, Invitrogen).
Icam 2 from rat
Manufactured by BD
ICAM-2 from rat is a laboratory reagent used for research purposes. It is a cell surface glycoprotein involved in cell-cell adhesion.
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2 protocols using icam 2 from rat
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Choroidal Vasculature Flat Mount Staining
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Choroidal Vasculature Flat Mount Staining
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Hematoxylin & Eosin staining was performed as described (Wang et al., 2008 (link)). For flat mount staining of the choroidal vasculature, pigment bleaching in RPE cells and choroidal melanocytes was performed as described (Kim and Assawachananont, 2016 ). Briefly, pigmented eyes were fixed in 4% paraformaldehyde PBS for 30 min to 2 h. After washing with PBS three times, the cornea and lens were removed before 3% hydrogen peroxide bleaching at 55 °C for 30 min to 2 h depending on the stage, or at 4 °C for several days. Immunofluorescence staining was performed using standard procedures. Primary antibodies used include: Beta-galactosidase (LacZ) from chicken (1:300, Abcam), Endomucin from rat (1:200, Santa Cruz Biotechnology), PECAM-1 from rat (1:50, BD Pharmingen), ICAM-2 from rat (1: 200, BD Pharmingen). These three EC markers, Endomucin, PECAM-1 and ICAM-2, were used interchangeably, but some antibodies worked better than others due to different sample processing. Secondary antibodies include: goat antichicken (or rat) AlexaFluor 488 (1:500, Invitrogen) and goat anti-rat AlexaFluor 594 (1:500, Invitrogen).
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