Figure 1.
The isolated inclusion bodies obtained from 6 g of IBD+ liver.
The arrows showing pelleted inclusion bodies in 1% Sarkosyl after centrifugation.
Figure 2.
The H&E stained IB prep on a microscopic slide.
The semi-purified inclusion bodies were used to immunize mice for antibody production. Bar = 20 µm.
Figure 3.
The resolved IB preps and liver homogenates on a NuPAGE.
Ten microliters of protein were loaded on each lane. Lane 1 to 3 are three different IB preps obtained from three IBD positive boa constrictors. Lane 4 is the liver homogenate from the same boa as lane 3. Lane 5 and 6 are liver homogenates from two IBD negative boas. Lane 7 is the IB prep derived from an IBD negative boa, which no pellet were left after incubating with 1% sarkosyl. Lane 8 is HB served as a blank control. The IB prep from #08-76 showed a major intense band approximately at 68 KDa (arrow), showing that the IBDP is concentrated in the IB prep compared to the liver homogenate (Lane 4). M. Molecular weight marker (Invitrogen, Mark 12).
Figure 4.
Immuno-gold labeling of IBD affected liver and kidney from boa constrictors.
A. gold particles conjugated with purified anti-IBDP MAB labeling an inclusion body within liver of a boa constrictor. Bar = 2 µm. B. gold particles conjugated with polyclonal antibody (Mouse 1 serum) labeling an inclusion body within kidney of a boa constrictor. Bar = 1 µm. C. gold particles conjugated with polyclonal antibody (Mouse 1 serum) labeling an inclusion bodies within kidney of a boa constrictor. Bar = 2 µm.
Figure 5.
Pancreas of an IBD positive boa constrictor stained with standardized IHC condition.
The tissue was fixed and stained under standardized IHC staining conditions using NovaRED as substrate. The negative control was stained with non-specific mouse antibody. The cell nucleus stained dark blue with hematoxylin, and the inclusion bodies are indicated by arrows. A. The inclusion bodies were not stained in the negative control slide. Bar = 20 µm.. B. The inclusion bodies stained dark red by anti-IBDP MAB. Bar = 20 µm. C. The inclusion bodies were not stained in the negative control slide. Bar = 100 µm. D. The positive stained pancreas by anti-IBDP MAB. Bar = 100 µm.
Figure 6.
Mean IHC score of liver, kidney, and pancreas over fixation time in 10% NBF.
Figure 7.
Differences in IHC staining between Trilogy and double AR treatment on prolong fixed tissues.
Tissue fixed up to 50 days (Block 9) in 10% NBF was stained with standardized Trilogy AR treatment (left) and double AR treatment (right). The inclusion bodies are indicated by arrows. Bar = 40 µm. A. In pancreas, the staining of inclusion bodies decreased when double AR was used. B. In kidney, the staining of inclusion bodies decreased when double AR was used. C. In liver, the staining of inclusion bodies improved when double AR was used.