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Site Directed Mutagenesis (SDM) Human

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Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

RNA siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line) SIRT1

Get tips on using siGENOME Human FTO (79068) siRNA - Individual to perform siRNA / miRNA gene silencing Human - SHSY5Y FTO

Products Horizon Discovery Ltd. siGENOME Human FTO (79068) siRNA - Individual

Get tips on using Human Syndecan-1 DuoSet ELISA to perform ELISA Human - SDC1

Products R&D Systems Human Syndecan-1 DuoSet ELISA

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

RNA siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line) HSPA5 (GRP78)

Get tips on using Human SDC1 / Syndecan-1 ELISA Kit to perform ELISA Human - SDC1

Products Sigma-Aldrich Human SDC1 / Syndecan-1 ELISA Kit

Get tips on using esiRNA human PTPN3 (esiRNA1) to perform siRNA / miRNA gene silencing Human - A2780 PTPN3

Products Sigma-Aldrich esiRNA human PTPN3 (esiRNA1)

Get tips on using ON-TARGETplus Human THBS2 siRNA to perform siRNA / miRNA gene silencing Human - Aortic smooth muscle cell TSP-2

Products Dharmacon ON-TARGETplus Human THBS2 siRNA

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

Proteins ELISA Human SDC1

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Cellular assays TUNEL assay cell type HeLa cells human cervical cancer

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Cellular assays TUNEL assay cell type PANC-1 human pancriatic cancer

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