Dab abc detection ihc kit

Localization of S1 protein expression was investigated in both kidney and lung tissues from all groups at week 6 after the first vaccination. IHC using rabbit-specific HRP/DAB (ABC) detection IHC kit (Abcam, Cambridge, UK) was adapted, with using antinovel coronavirus spike protein S1 polyclonal antibodies (1:1000; Sino-Biological, PA, USA). A 5-µm thick section of formalin was fixed, and paraffin-embedded tissues were cut and loaded on specially coated slides. The prepared slides were put in an oven at 70°C for 30 min followed by 15 min in a xylene solution for deparaffinization. The sections were dried and surrounded by liquid blocker (Dako pen). The antigen retrieval (pretreatment) step was executed by immersing the sections in 1× reveal Decloaker reagent (Biocare Medical, Pacheco, CA, USA) and heating in a microwave several times for 30 min then cool at room temperature for 15-20 min. After that, staining procedure was carried out by following the manufacturer’s instructions of the ABC kit. To eliminate the effect of internal renal biotin, 0.05% avidin was applied (Sigma-Aldrich, St. Louis, MO, USA) only on kidney sections for 15 min at room temperature [30 (link),31 (link)]. The immunoreactivity of the expressed S1 protein was visualized using a light microscope at different magnifications.

Proliferative-phase endometrial tissues from 10 patients with RIF and 10 FER controls were collected and washed with PBS immediately. Tissues were fixed in 4% paraformaldehyde, paraffin-embedded, and serially sectioned. Immunostaining was performed according to the instruction of the mouse and rabbit specific HRP/diaminobenzidine (DAB) (ABC) Detection IHC Kit (ab64264, Abcam). The slides were incubated with primary antibody against NSD2 (1:100, ab223694, Abcam), H3K36me2 (1:1000, ab176921, Abcam), minichromosome maintenance complex component 7 (MCM7) (1:500, 11225-1-AP, Proteintech), or Rabbit IgG (1:500, #3900, Cell Signaling Technology) diluted in PBS overnight at 4°C. The primary antibodies used in the study were all commercial antibodies reported by other empirical studies (Zhu et al. 2019 (link), Dai et al. 2020 (link), Ganig et al. 2021 (link), Acke et al. 2022 (link)). For each immunohistochemical run, a slice from each group was randomly selected as the negative control. The negative control was tested by incubation with immunoglobulin G (IgG) from the corresponding species. Next day, they were incubated with secondary antibodies, stained with DAB, and counterstained with hematoxylin. The slides were then dehydrated, cleared, and mounted. The images were captured under a microscope. See Supplementary methods for the immunohistochemical scoring method.