Protein Expression Eukaryotic cells HEK293

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Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - Human eutopic endometrial stromal cells

Products Sigma-Aldrich CelLytic™ NuCLEAR™ Extraction Kit

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

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

DNA Site Directed Mutagenesis (SDM) Human Point mutation PC-3 Speckle-Type POZ protein (SPOP)

Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform Microarray Human - Endometrial stromal cells Expression array

Products Thermo Fisher Scientific GeneChip® Human Genome U133 Plus 2.0 Array

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

DNA Microarray Comperative genomic hybridization Human Blood cells

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Circumvallate papillae

Products Sigma-Aldrich RIPA Buffer

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - 3T3-L1

Products Sigma-Aldrich RIPA Buffer

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - THP-1

Products Sigma-Aldrich RIPA Buffer

Get tips on using Trichloroacetic acid to perform Protein isolation Mammalian cells - THP-1

Products Sigma-Aldrich Trichloroacetic acid

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - CHO-K1

Products Sigma-Aldrich RIPA Buffer

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