Western blot protocol for detecting ATP10B in mouse/rat brain v1

Author(s):  
María Sanchiz Calvo ◽  
eduard.bentea not provided ◽  
Veerle Baekelandt

Protocol for detection of ATP10B in rat and mouse brain tissue by Western blotting

2019 ◽  
Vol 15 (3) ◽  
pp. 251-257
Author(s):  
Bahareh Sadat Yousefsani ◽  
Seyed Ahmad Mohajeri ◽  
Mohammad Moshiri ◽  
Hossein Hosseinzadeh

Background:Molecularly imprinted polymers (MIPs) are synthetic polymers that have a selective site for a given analyte, or a group of structurally related compounds, that make them ideal polymers to be used in separation processes.Objective:An optimized molecularly imprinted polymer was selected and applied for selective extraction and analysis of clozapine in rat brain tissue.Methods:A molecularly imprinted solid-phase extraction (MISPE) method was developed for preconcentration and cleanup of clozapine in rat brain samples before HPLC-UV analysis. The extraction and analytical process was calibrated in the range of 0.025-100 ppm. Clozapine recovery in this MISPE process was calculated between 99.40 and 102.96%. The limit of detection (LOD) and the limit of quantification (LOQ) of the assay were 0.003 and 0.025 ppm, respectively. Intra-day precision values for clozapine concentrations of 0.125 and 0.025 ppm were 5.30 and 3.55%, whereas inter-day precision values of these concentrations were 9.23 and 6.15%, respectively. In this study, the effect of lipid emulsion infusion in reducing the brain concentration of drug was also evaluated.Results:The data indicated that calibrated method was successfully applied for the analysis of clozapine in the real rat brain samples after administration of a toxic dose to animal. Finally, the efficacy of lipid emulsion therapy in reducing the brain tissue concentration of clozapine after toxic administration of drug was determined.Conclusion:The proposed MISPE method could be applied in the extraction and preconcentration before HPLC-UV analysis of clozapine in rat brain tissue.


1972 ◽  
Vol 247 (8) ◽  
pp. 2322-2327
Author(s):  
Frederico A. Cumar ◽  
John F. Tallman ◽  
Roscoe O. Brady

1970 ◽  
Vol 245 (6) ◽  
pp. 1388-1393
Author(s):  
Michael P. Lerner ◽  
Terry C. Johnson

1989 ◽  
Vol 9 (1) ◽  
pp. 87-95 ◽  
Author(s):  
Michihiro Kirikae ◽  
Mirko Diksic ◽  
Y. Lucas Yamamoto

We examined the rate of glucose utilization and the rate of valine incorporation into proteins using 2-[18F]fluoro-2-deoxyglucose and L-[1-14C]-valine in a rat brain tumor model by quantitative double-tracer autoradiography. We found that in the implanted tumor the rate of valine incorporation into proteins was about 22 times and the rate of glucose utilization was about 1.5 times that in the contralateral cortex. (In the ipsilateral cortex, the tumor had a profound effect on glucose utilization but no effect on the rate of valine incorporation into proteins.) Our findings suggest that it is more useful to measure protein synthesis than glucose utilization to assess the effectiveness of antitumor agents and their toxicity to normal brain tissue. We compared two methods to estimate the rate of valine incorporation: “kinetic” (quantitation done using an operational equation and the average brain rate coefficients) and “washed slices” (unbound labeled valine removed by washing brain slices in 10% thrichloroacetic acid). The results were the same using either method. It would seem that the kinetic method can thus be used for quantitative measurement of protein synthesis in brain tumors and normal brain tissue using [11C]-valine with positron emission tomography.


2008 ◽  
Vol 50 (4) ◽  
pp. 759-767 ◽  
Author(s):  
Dixie-Ann Persaud-Sawin ◽  
Samantha Lightcap ◽  
G. Jean Harry
Keyword(s):  

2002 ◽  
Vol 323 (3) ◽  
pp. 207-210 ◽  
Author(s):  
Noriko Mochizuki-Oda ◽  
Yosky Kataoka ◽  
Yilong Cui ◽  
Hisao Yamada ◽  
Manabu Heya ◽  
...  

1993 ◽  
Vol 60 (5) ◽  
pp. 1639-1649 ◽  
Author(s):  
H.-P. Lipp ◽  
D. P. Wolfer ◽  
W. X. Qin ◽  
C. B. Klee ◽  
C. W. Heizmann

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