Cy5 affinipure donkey anti rabbit igg

Placentas were fixed in 4% paraformaldehyde in phosphate buffered saline overnight and then processed into paraffin blocks. Midline cross-sections were cut 5 μm thick. To determine phenotypical differences of the placentas, H and E staining was performed by the UNC Animal Histopathology and Lab Medicine Core and then imaged using a Zeiss Axio Image.A2. For immunofluorescence identification of uterine natural killer (uNK) cells, dendritic (CD11c) cells, and smooth muscle actin (αSMA), placentas were de-paraffinized using a clearing reagent and then rehydrated using a series of ethanol washes. Following antigen retrieval, sections were then blocked with 5% normal donkey serum and incubated overnight at 4°C with the primary antibody CD11c (Proteintech, 17342-1-AP;1:50) or anti-α-Smooth Muscle Actin (Sigma-Aldrich, clone 1A4, A2547; 1:100) and incubated the next day with secondary antibodies including Cy™5 AffiniPure donkey anti-rabbit IgG (Jackson ImmunoResearch; 711-175-152; 1:200), Cy™2 AffiniPure donkey anti-rabbit IgG (Jackson ImmunoResearch; 711-225-152; 1:200), FITC-conjugated DBA lectin (Sigma-Aldrich; L9142; 1:200), and Hoechst (Sigma-Aldrich; B1155; 1:500). Images were acquired using an IX83 Olympus microscope with a Hamamatsu digital camera with cellSens dimension software (Olympus).