siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - mouse pancreatic stellate cells

Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells

Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

Get tips on using ONE-Glo™ Luciferase Assay System to perform Reporter gene assay luciferase - BHK-21 baby hamster kidney 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.

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

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.

Get tips on using Senescence Cells Histochemical Staining Kit to perform Cell cycle assay mouse - L929

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.