Detection of two Prorocentrum species using sandwich hybridization integrated with nuclease protection assay

Harmful Algae ◽  
2006 ◽  
Vol 5 (3) ◽  
pp. 300-309 ◽  
Author(s):  
Qingsong Cai ◽  
Rongxiu Li ◽  
Yu Zhen ◽  
Tiezhu Mi ◽  
Zhigang Yu
Keyword(s):  
Protection Assay ◽  
Sandwich Hybridization ◽  
Nuclease Protection Assay

Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH)

Hydrobiologia ◽  
2006 ◽  
Vol 575 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Yu Zhen ◽  
Zhigang Yu ◽  
Qingsong Cai ◽  
Tiezhu Mi ◽  
Rongxiu Li
Keyword(s):  
Protection Assay ◽  
Sandwich Hybridization ◽  
Nuclease Protection Assay

S1 Nuclease Protection Assay

1986 ◽  
pp. 276-284 ◽  
Author(s):  
Leonard G. Davis ◽  
Mark D. Dibner ◽  
James F. Battey
Keyword(s):  
Protection Assay ◽  
S1 Nuclease ◽  
Nuclease Protection Assay

scholarly journals Regulation of thrombopoietin levels by c-mpl-mediated binding to platelets

Blood ◽  
1996 ◽  
Vol 87 (6) ◽  
pp. 2154-2161 ◽  
Author(s):  
PJ Fielder ◽  
AL Gurney ◽  
E Stefanich ◽  
M Marian ◽  
MW Moore ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Blood Cells ◽  
Platelet Rich Plasma ◽  
Plasma Concentrations ◽  
Mrna Levels ◽  
Protection Assay ◽  
S1 Nuclease ◽  
Intravenous Injections ◽  
Nuclease Protection Assay ◽  
Specific Uptake

The involvement of platelets and the c-mpl receptor in the regulation of thrombopoietin (TPO) plasma concentrations and tissue mRNA levels was investigated in both normal mice and mice defective in c-mpl (c-mpl- /-). Although c-mpl-/- mice have fewer platelets and higher plasma TPO activity than normal mice, there was no increase in TPO mRNA levels as measured by an S1 nuclease protection assay. After the intravenous injection of 125I-TPO, specific uptake of radioactivity by the spleen and blood cells was present in the normal mice, but absent in the c-mpl- /- mice. Platelet-rich plasma (PRP) from normal mice was able to bind and internalize 125I-TPO, whereas PRP from c-mpl-/- mice lacked this ability. Analysis of 125I-TPO binding to normal PRP indicated that binding was specific and saturable, with an approximate affinity of 560 pmol/L and 220 receptors per platelet. PRP from normal mice was also able to degrade 125I-TPO into lower molecular weight fragments. After the intravenous injections, c-mpl-/- mice cleared a dose of 125I-TPO at a much slower rate than did normal mice. Injection of washed platelets from normal mice into c-mpl-/- mice resulted in a dramatic, but transient, decrease in plasma TPO levels. These data provide evidence that platelets regulate plasma TPO levels via binding to the c-mpl receptor on circulating platelets.

scholarly journals The mitochondrial housekeeping gene 16S is inappropriate as an internal standard in comparative studies of rare mitochondrial transcripts using S1-nuclease protection assays

2010 ◽  
Vol 1 (1) ◽  
pp. 2
Author(s):  
Sandra Ebert ◽  
Line Breumlund ◽  
Roland Nau ◽  
Uwe Michel
Keyword(s):  
Internal Standard ◽  
Human Neuroblastoma Cell ◽  
Protection Assay ◽  
Human Neuroblastoma ◽  
Rnase Protection ◽  
S1 Nuclease ◽  
Total Rna ◽  
16S Rna ◽  
Nuclease Protection Assay ◽  
Mitochondrial Transcripts

The analysis of rare mitochondrial transcripts derived from the L-strand of the mitochondrial genome requires a sensitive method such as the S1-nuclease protection assay. We examined whether the ribosomal mitochon­drial transcript 16S is suitable as an internal standard in a multiplex S1-nuclease protection assay for the measurement of different mitochondrial transcripts. For reliable quantification of rare mitochondrial transcripts with the RNase protection assay, a minimum of 2 μg of total RNA is necessary. Standard curves of 16S RNA produced with total RNA from human kidney, liver, brain, and a human neuroblastoma cell line (SH-SY5Y) revealed dose-response relationships that were saturated already at less than 0.5 μg of total RNA. Therefore, 16S is inappropriate as an internal standard for analyzing mitochondrial transcripts with RNase protection assays when more than 0.5 μg of total RNA have to be analyzed.

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