The study included 36 males and 14 females, and the breeds presented included Netherland dwarfs (n = 8), Holland lop-eared rabbits (n = 8), mini-rabbits (n = 10), and other breeds (n = 24).
The proximity ligation assay probe-coupled antibodies used were anti-goat plus, anti-goat minus, anti-rabbit plus, anti-rabbit minus, anti-mouse plus and anti-mouse minus.
When I was 9, I started earning money by breeding mini lop rabbits and selling them through the local newspaper.
To test for the effects of drugs on the swimming behavior of Ciona larvae, video recordings of larval movements were made at five-minute intervals from individual larvae in filtered seawater with a high speed video (Fast-Cam Rabbit mini 2, Photoron Tokyo, Japan) at 400 to 500 fps mounted on a binocular microscope and stored for later analysis.
To detect primary antibodies with the in situ proximity ligation assay (PLA), the PLA probes mouse PLUS and rabbit MINUS (Abnova, UK) were added at a 1∶5 dilution in antibody dilution buffer (Olink Bioscience, Sweden) for 60 min at 37°C.
After washing, the slides were incubated with Duolink PLA Rabbit MINUS and PLA Mouse PLUS proximity probes (Olink Bioscience) and a proximity ligation was performed using the Duolink Detection Reagent Kit (Olink Bioscience) according to the manufacturer's protocol.
pKT2-PGKi contains the human PGK promoter, a kind gift of Dr. Scott McIvor (University of Minnesota) in front of the mini-intron, MCS, and rabbit beta-globin 3'UTR found in mini-CAGs.
No notable differences were found in the mini-pig, monkey, and rabbit mechanical and biochemical properties, suggesting that these species, despite morphological differences, may serve as suitable animal models for studying structure-function relationships of the human facet joint.
PLA secondary probes anti-rabbit MINUS and anti-mouse PLUS (Duolink kit, Sigma), containing the secondary antibodies conjugated with oligonucleotides, were diluted 1 5 in Duolink antibody diluent.
After washing with TBS-T2 (TBS with 0.1% Triton X-100), the PLA probes anti-rabbit MINUS (Olink; 92005) and anti-mouse PLUS (Olink; 92001) were added at 1 5 in blocking solution for 1 h at room temperature (22°C).
After 1 hr primary antibody incubation at room temperature, the PLA probe anti-mouse Plus and anti-rabbit Minus were diluted in PBS containing 1% BSA and added to cells for 1 hr at 37°C in a humidified chamber.
