Tissues were fixed for 12–24 h in 10% formalin, embedded in paraffin, and sectioned. Sections were incubated overnight at 4°C with rat anti-Mac2 (CL8942AP; Cedarlane). Biotinylated rabbit anti-rat (E0467; Dako) was used as secondary antibody. The ABC peroxidase kit (Pierce) was used for immunodetection, and sections were counterstained in Mayer’s hematoxylin.
E0467
Manufactured by Agilent Technologies
The E0467 is a laboratory equipment product from Agilent Technologies. It serves as a core component for various analytical and testing applications. The device's primary function is to provide precise and reliable measurements, though its specific intended use is not detailed here.
Automatically generated - may contain errors
Lab products found in correlation
Cl8942ap, by Cedarlane (2 mentions)
Ab13524, by Abcam (1 mentions)
Z0334, by Agilent Technologies (1 mentions)
L5391, by Merck Group (1 mentions)
Eclipse 90i optical microscope, by Nikon (1 mentions)
Sc 19992, by Santa Cruz Biotechnology (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
3 protocols using e0467
1
Macrophage Immunohistochemistry Protocol
2
Quantitative Immunohistochemical Analysis of Lysosomal Markers
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Formalin-fixed, paraffin-embedded tissue sections were incubated overnight at 4°C with rat anti-LAMP1 (SC-19992, Santa Cruz Biotechnology, 1:100), rat anti-LAMP2 (ab13524, Abcam, 1:500), rabbit anti-LIMP2 (NB400; Novus Biologicals, 1:100), rabbit anti-GFAP (Z0334, Dako Cytomation, 1:1000) and BSI-B4 lectin (L5391, Sigma, 1:100). For bright-field immunostaining, biotinylated rabbit anti-rat IgG (E0467, Dako, 1:300) and biotinylated goat anti-rabbit IgG (31820, Vector Laboratories, 1:300) were used as secondary antibodies. Bright-field sections were stained with 3,3-diaminobenzidine (Sigma) and counterstained with haematoxylin. Images were obtained with an Eclipse 90i optical microscope (Nikon). LIMP2, LAMP2, GFAP and BSI-B4 signals were quantified with the NIS-Elements Advanced Research 2.20 software (Nikon) in 3-5 20× images of each brain region of each animal using the same signal threshold settings for all images. The percentage positive area of each image was then calculated.
3
Immunohistochemical Analysis of Adipocytes
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Tissues were fixed for 12-24 h in 10% formalin, embedded in paraffin and sectioned. Sections were incubated overnight at 4ºC with rat anti-Mac2 antibody (CL8942AP; Cedarlane). Biotinylated rabbit anti-rat (E0467; Dako) was used as secondary antibody. The ABC peroxidase kit (Pierce) was used for immunodetection, and sections were counterstained in Mayer's hematoxylin. Morphometric analysis of adipocyte size was performed in WAT sections stained with hematoxylin-eosin as previously described [34] . A minimum of four animals per group was used and at least 250 adipocytes per animal were analyzed.
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