scholarly journals Electrophoretic Mobility Shift Assay and Dimethyl Sulfate Footprinting for Characterization of G-Quadruplexes and G-Quadruplex-Protein Complexes

2019 ◽  
pp. 201-222 ◽  
Author(s):  
Buket Onel ◽  
Guanhui Wu ◽  
Daekyu Sun ◽  
Clement Lin ◽  
Danzhou Yang
Keyword(s):  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Protein Complexes ◽  
Dimethyl Sulfate ◽  
Mobility Shift ◽  
G Quadruplex ◽  
Mobility Shift Assay

Evaluation of electrophoretic mobility shift assay as a method to determine plasma protein binding of siRNA

Bioanalysis ◽  
2019 ◽  
Vol 11 (21) ◽  
pp. 1927-1939 ◽  
Author(s):  
Carrie Rocca ◽  
Sean Dennin ◽  
Yongli Gu ◽  
Joohwan Kim ◽  
Samantha Chigas ◽  
...  
Keyword(s):  
Protein Binding ◽  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Small Interfering Rnas ◽  
Mobility Shift ◽  
Mobility Shift Assay

Aim: The electrophoretic mobility shift assay (EMSA) was evaluated as an alternative to ultrafiltration (UF) to assess plasma protein binding (PPB) of small interfering RNAs (siRNA) and antisense oligonucleotides (ASO). Results & methodology: EMSA analysis showed that PPB depended on siRNA and plasma concentration. Conversely, when analyzed by ultrafiltration, siRNA bound the filtration device nonspecifically and PPB remained >98% across physiologically relevant siRNA concentrations. Using EMSA, siRNA exhibited charge-based interactions with plasma proteins, while ASO remained highly bound to plasma proteins or albumin in the presence of 500 mM salt. Conclusion: PPB characteristics of siRNA and ASO can be distinguished using EMSA. Characterization of siRNA PPB by EMSA enhances our knowledge of siRNA absorption, distribution, metabolism and excretion and advanced development of RNA interference therapeutics.

scholarly journals Characterization of Tubulin Isotype-Specific Antibodies by Electrophoretic Mobility Shift Assay

BioTechniques ◽  
1998 ◽  
Vol 25 (6) ◽  
pp. 940-942 ◽  
Author(s):  
Monica L. Fanarraga ◽  
Kerman Aloria ◽  
Jesus Avila ◽  
Juan C. Zabala
Keyword(s):  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Mobility Shift ◽  
Specific Antibodies ◽  
Tubulin Isotype ◽  
Mobility Shift Assay

A fluorescence based non-radioactive electrophoretic mobility shift assay

2000 ◽  
Vol 78 (2) ◽  
pp. 163-170 ◽  
Author(s):  
K Ruscher ◽  
M Reuter ◽  
D Kupper ◽  
G Trendelenburg ◽  
U Dirnagl ◽  
...  
Keyword(s):  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Mobility Shift ◽  
Mobility Shift Assay

scholarly journals Human replication-dependent histone H3 genes are activated by a tandemly arranged pair of two CCAAT boxes

2004 ◽  
Vol 384 (2) ◽  
pp. 317-326 ◽  
Author(s):  
Heiner KOESSLER ◽  
Joerg KAHLE ◽  
Christa BODE ◽  
Detlef DOENECKE ◽  
Werner ALBIG
Keyword(s):  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Histone H3 ◽  
Maximal Activity ◽  
Significant Loss ◽  
Tata Box ◽  
Mobility Shift ◽  
Nuclear Factor Y ◽  
Binding Factor ◽  
Mobility Shift Assay

We have analysed the transcriptional regulation of the human histone H3 genes using promoter deletion series, scanning mutagenesis, specific mutagenesis and electrophoretic mobility-shift assay experiments. The promoters of five of the six examined histone H3 genes showed near-maximal activity at lengths of 133–227 bp: H3/d 198 bp, H3/h 147 bp, H3/k 133 bp, H3/m 227 bp, H3/n 140 bp (exception H3/i). To search for functional cis-elements within these regions, we performed scanning mutagenesis of the two histone H3 promoters H3/k and H3/m. Mutagenesis revealed that the functional framework of the histone H3 promoters consists of a TATA box and two tandemly arranged CCAAT boxes in relatively fixed positions. Alterations of the distance between the CCAAT boxes and of the distance between the CCAAT boxes and the TATA box resulted in significant loss of activity. In electrophoretic mobility-shift assay experiments, the factor CBF (CCAAT-binding factor)/NF-Y (nuclear factor-Y) bound to isolated CCAAT boxes of the H3/k promoter. This suggests that an initiation complex is formed on the histone H3 promoter that has a defined structure and limited flexibility, consisting of two molecules of CBF/NF-Y and further (general or specific) transcription factors.

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