CRISPR Rat Deletion INS-1 832/13

Get tips on using PM-1 Subcutaneous Preadipocyte Medium to perform Stem cell culture media hAdipose derived stem cells

Get tips on using Anti-Beclin 1 (Human) pAb to perform Autophagy assay cell type - Mouse white adipose tissue

Get tips on using Cell Proliferation Reagent WST-1 to perform Cell cytotoxicity / Proliferation assay cell type - L-02

Get tips on using Ccl2 siRNA to perform siRNA / miRNA gene silencing Rat - Neuronal cells MCP-1

Get tips on using SNAI 1 siRNA and shRNA Plasmids (h) to perform siRNA / miRNA gene silencing Human - MDA-MB-468 SNAI 1

Get tips on using SNAI 1 siRNA and shRNA Plasmids (h) to perform siRNA / miRNA gene silencing Human - MDA-MB-231 SNAI 1

Get tips on using Monoclonal Anti-Caveolin-1 antibody produced in mouse to perform Western blotting Caveolin-1

Reporter gene assays are designed to test the regulation of the expression of a gene of interest. This is usually done by linking the promoter of the gene of interest with a gene such as a firefly luciferase, which can be easily detected by addition of luciferin that leads to an enzymatic reaction to produce luminescence. The enzymatic reaction can be correlated to the expression of the gene of interest. Another luciferase gene that can be used is Renilla luciferase. For an appropriate luciferase assay: 1. the reporter should express uniformly in all cells, 2. specifically respond to effectors that the assay intends to monitor, 3. have low intrinsic stability to quickly reflect transcriptional dynamics. It is important to have an equal number of cells plated in each testing condition to avoid any incorrect readouts. Reporter assays could be single or dual reporter assays. The reporter could be both luciferases. Most dual-luciferase assays involve adding two reagents to each sample and measuring luminescence following each addition. Adding the first reagent activates the first luciferase reporter reaction; adding the second reagent extinguishes first luciferase reporter activity and initiates the second luciferase reaction. Dual-luciferase assays have some advantages, including 1. reduces variability, 2. reduces background, 3. normalizes differences in transfection efficiencies between samples.

Get tips on using Human Chitinase 3-like 1 DuoSet ELISA to perform ELISA Human - Chitinase-3-Like Protein-1 (CHI3L1) or YKL-40

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.