Immunohistochemistry 53BP2 phospho (ser-25)

Get tips on using ON-TARGETplus Human SAMHD1 (25939) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - HFF1 SAMHD1

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - H1299 GAPDH

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - A549 GAPDH

Get tips on using ON-TARGETplus Rat Smad3 (25631) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad3

Get tips on using ON-TARGETplus Rat Hspa5 (25617) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - AR42J Grp78/Hspa5

Get tips on using ON-TARGETplus Rat Pld2 (25097) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Pld2

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - Calu-3 GAPDH

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.

Get tips on using RNAprotect Tissue Reagent (250 ml) to perform RNA stabilization Sorted 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.