Immunofluorescence staining was performed for each tumor specimen using antibodies against CD31 (anti-mouse, 1:200) (Millipore, Billerica, MA), glial fibrillary acidic protein (GFAP) (anti-rabbit, 1:1000) (Abcam, Cambridge, UK), and aquaporin 4 (AQ4) (anti-rabbit, 1:200) (Millipore). AQ4 is a marker of astrocytic foot processes and has been previously used as a reliable measure of astrocyte-endothelial cell relationship integrity [6 (link),7 (link)]. Double immunofluorescence staining was performed for CD31 and GFAP; and CD31 and AQ4, using a protocol described previously [5 (link)]. The following secondary antibodies were used at 1:1000 dilution:donkey serum anti-mouse IgG absorbing at a wavelength of 568 nm (Life Technologies, Carlsbad, CA) and donkey serum anti-rabbit IgG absorbing at a wavelength of 488 nm (Life Technologies). Immunofluorescent images were captured at ×40 magnification using a Leica (Wetzlar, Germany) LSM 510 confocal microscope, while H and E images were obtained at ×40 magnification using a Nikon (Tokyo, Japan) Eclipse Ci microscope with a Nikon DS-Fi2 camera.
Donkey serum anti rabbit igg
Manufactured by Thermo Fisher Scientific
Donkey serum anti-rabbit IgG is a laboratory reagent used for the detection and quantification of rabbit immunoglobulin G (IgG) in various applications such as immunoassays and Western blotting. It is derived from donkey serum and contains antibodies that specifically bind to rabbit IgG, allowing for the identification and measurement of rabbit IgG in samples.
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2 protocols using donkey serum anti rabbit igg
1
Quantifying Astrocyte-Endothelial Interactions in Tumor Specimens
2
Quantifying Astrocyte-Endothelial Interactions in Tumor Specimens
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Immunofluorescence staining was performed for each tumor specimen using antibodies against CD31 (anti-mouse, 1:200) (Millipore, Billerica, MA), glial fibrillary acidic protein (GFAP) (anti-rabbit, 1:1000) (Abcam, Cambridge, UK), and aquaporin 4 (AQ4) (anti-rabbit, 1:200) (Millipore). AQ4 is a marker of astrocytic foot processes and has been previously used as a reliable measure of astrocyte-endothelial cell relationship integrity [6 (link),7 (link)]. Double immunofluorescence staining was performed for CD31 and GFAP; and CD31 and AQ4, using a protocol described previously [5 (link)]. The following secondary antibodies were used at 1:1000 dilution:donkey serum anti-mouse IgG absorbing at a wavelength of 568 nm (Life Technologies, Carlsbad, CA) and donkey serum anti-rabbit IgG absorbing at a wavelength of 488 nm (Life Technologies). Immunofluorescent images were captured at ×40 magnification using a Leica (Wetzlar, Germany) LSM 510 confocal microscope, while H and E images were obtained at ×40 magnification using a Nikon (Tokyo, Japan) Eclipse Ci microscope with a Nikon DS-Fi2 camera.
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