Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Get tips on using ON-TARGETplus Human ITGB4 (3691) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - DU145 ITGB4
Get tips on using FITC Rat Anti-Human CD49f to perform Flow cytometry Anti-bodies Human - CD49f/ITGA6
Get tips on using ON-TARGETplus Human ITGB4 (3691) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - CAL-27 ITGB5
Get tips on using Live and Dead Cell Assay (Abcam) to perform Live / Dead assay mammalian cells - rabbit bone marrow mesenchymal stem cells
Get tips on using ON-TARGETplus Human ITGB3 (3690) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MDA-MB-231 β3 integrin/ITGB3
Get tips on using ON-TARGETplus Human ITGB1 (3688) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MDA-MB-231 β1 integrin/ITGB1
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.
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.
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.
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