rna-isolation-purification-bacteria-gram-negative-salmonella-enterica

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I would like to regulate the expression of a gene and in order to do that, I have purchased specific siRNA. After optimizing my transfection protocol and using electroporation I have achieved a 60-70% reduction of the gene of interest. However, I cannot observe a significant reduction of mRNA expression but only a reduction of protein. What might be the problem? Could the problem be in my cell treatment method?

Discussions siRNA/RNAi/miRNA transfection human

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Proteins Western blotting tubulin gamma

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Get tips on using γ Tubulin Antibody (TU-30): sc-51715 to perform Western blotting tubulin gamma

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Get tips on using Monoclonal Anti-γ-Tubulin antibody produced in mouse to perform Western blotting tubulin gamma

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The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

DNA DNA methylation profiling Gene specific profiling MG-63 RANKL

Get tips on using Rn_LOC311846_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - Astrocytes LRRC8A

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Get tips on using Rn_Sod2_4 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - PC12 SOD2

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