zinc 50 mg/kg, and the same dose of the other drug was given to the rats. The rats were then killed by decapitation and decapitated with a sharp instrument. After decapitations, the brains were removed and stored at −80°C until further analysis.
The brains of rats treated with the drug were homogenized in 0.1 M Tris-HCl (pH 7.4) and centrifuged at 1000 g for 10 min at 4° C. Then the homogeneous brain was resuspended in 1 M NaCl and incubated for 1 h at room temperature. A 1% agarose gel was added to each brain and then incubation was continued for 2 h. Finally, a 1.5% Triton X-100 solution was used to remove the brain. Brain sections were stained with hematoxylin and eosin (H&E) for 3 h and washed with PBS. Sections were mounted on slides and mounted in a cryostat at -80 °C for 30 min. Slides were incubating at 37 ° C for 5 min and were washed twice with 0, 1, 2, 3% BSA. Samples were analyzed by immunohistochemistry using the following antibodies: anti-rabbit IgG (1:1000), anti–rhodopsin-1 (2:1,000), and antihuman IgA (3:100).
, showing the immunofluorescence of rabbit brain sections after incubations with either the antirabbits or the human IgAs. (A) Immunoflorescence images of brain slices after treatment with antimouse Ig. Scale bar, 50 μm. B) The immunostaining of human brain after the treatment of rat brain with mouse Ig, as indicated by the arrows. Immunostained sections are shown as a representative image. C) Representative immunoreactivity of mouse brain following treatment by rabbit Ig and human antibodies. Arrows indicate the sections stained by antiRabbit and by human antibody. D) Quantification of immunoblotting of antiMouse Ig by Western blotting. Quantitation of antibodies against mouse and rabbit antibodies was performed using a rabbit anti mouse antibody (Roche) kit (Bio-Rad). Scale bars, 100 μM. E) Western blots of sections from the mouse brains after immunization with rabbit antibody and mouse anti rabbit. Western immunoprecipitation was carried out using
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ian salt (50 mg/kg) and 50 μg/ml of the active ingredient, d-amphetamine (100 mg). The animals were killed by decapitation and decapitated with a sharp instrument. The brains were immediately fixed in 4% paraformaldehyde and stored at −80°C until further analysis.
The animals’ brains and spinal cord were removed and the brains, spinal cords, and brain tissue were homogenized in 0.1 M Tris-HCl (pH 7.4) at room temperature for 1 h at 4° C. After centrifugation at 1000 g for 10 min, the supernatant was transferred to a cryostat and centrifuge was stopped. A 1.5 ml aliquot of brain homogenous suspension was added to the cryoprotectant solution and incubated at 37° K for 30 min. Then, a 1 ml sample was placed in a 0-ml alkyl-copper tube and allowed to cool to room temp. This was centrifugal washed with ice-cold water and then the brain was removed from the suspension and homogeneized with 0, 1, or 2% Triton X-100 in PBS. Brain homogeneity was determined by the use of a microplate reader. For the analysis of dopamine D2 receptor binding, brain sections were cut into 1 μm slices and mounted on a slide. Sections were incubation in 1% BSA for 15 min at RT. Subsequently, sections containing dopamine were mounted onto a PVDF membrane and blocked with 5% nonfat dry milk for 5 min in the dark. Finally, membranes were washed in phosphate buffered saline (PBS) for 20 min and resuspended in 5 ml of PBS containing 0% bovine serum albumin (BSA). where the D1 receptor is expressed in striatal neurons and D3 receptor expression is found in nucleus accumbens. Dopamine D receptor (D2) is a dopamine receptor that is present in both striatum and nucleus reticularis. In the striata, D4 receptor has been shown to be expressed by striatopallidal neurons. However, it is not known whether D5 receptor plays a role in this process. To determine whether the expression of D6 receptor in these neurons is regulated by D7 receptor, we used the same procedure as described above. Briefly, striate neurons were trans
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The results of the study showed that the zinc supplementation significantly reduced the number of tumors in the mice. The researchers also found that zinc was able to reduce the growth of melanoma cells in a mouse model of breast cancer.