Peroxidase conjugated donkey anti goat igg

Phospholipid was purchased from Avanti (Shanghai, China); Cholesterol and glutaric anhydride were purchased from Aladdin (Shanghai, China); Sodium cholate was purchased from Tokyo Chemical Industry (Tokyo, Japan); Human apoA-I standard was purchased from Merck (Branchburg, NJ); FIV precipitate was kindly provided from Tonglu Biotechnology Co. Ltd. (Hangzhou, China); LCAT colorimetric assay kit was purchased from GenMed Scientifics Inc. (Arlington, MA); Goat anti-human apoA-I antibody and peroxidase-conjugated donkey anti-goat IgG were purchased from Abcam (Cambridge, UK). Mouse brain microvascular endothelial cells (bEND.3) and rat glioma cells (C6) were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). The rest were commercially qualified reagents.

Anti-ASC (Cell Signaling Technology, 67824, 1:1,000), anti-NLRP3 (Cell Signaling Technology, 15101, 1:1,000), anti-cleaved IL-1β (Cell Signaling Technology, 52718, 1:1,000), anti-pro–IL-1β (Cell Signaling Technology, 31202, 1:1,000), anti-DGAT1 (Gene-Protein Link, 1:1,000), anti–β-actin (Cell Signaling Technology, 3700, 1:2,000), anti-pro–caspase-1/cleaved caspase-1 p20 (AdipoGen Life Sciences, AG-20B-0042, 1:1,000), anti-NEK7 (Santa Cruz, sc-50756, 1:1,000), anti-GSTO1 (Proteintech, 15124-1-AP, 1:1,000), anti-GSH (ViroGen, 101-A, 1:1,000), anti-calnexin (LifeSpan Biosciences, LS-B9772-200, 1:1,000), anti-FACL4 (ABclonal, A20414, 1:1,000), peroxidase-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch, 111-035-144, 1:5,000), anti-mouse IgG (Jackson ImmunoResearch, 111-035-146, 1:5,000), peroxidase-conjugated donkey anti-goat IgG (Abcam, ab97110, 1:5,000), and Alexa Fluor 488–conjugated chicken anti-goat IgG (Invitrogen, 21467, 1:1,000) were used in our experiments.

The gonad–mesonephros complexes were fixed in modified Zamboni fixative containing 4% paraformaldehyde at 4°C for 24 hr, as described previously [9 (link),
22 (link)]. The specimens were dehydrated with an ethanol series followed by xylene and then embedded in paraffin. Then, 5-μm-thick sections at 25-μm
intervals were cut by a sliding microtome and placed on slide glasses that had been precoated with 2% 3-aminopropyltriethoxysilan (Shin-Etsu Chemical, Tokyo, Japan). The sections were then
stored at −18°C until use. After deparaffinization and hydration, the sections were stained with hematoxylin and eosin (HE) for general histology and measurement of MD diameter.
Immunohistochemical staining was carried out as described previously [9 (link), 22 (link)]. For the primary antibody, an
anti-AMH goat polyclonal antibody (1:12,000; sc-6886; Santa Cruz Biotechnology, Dallas, TX, USA) was used. For the secondary antibody, peroxidase-conjugated donkey anti-goat IgG (1:200,
ab97112; Abcam, Cambridge, UK) was used.