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.
Get tips on using BrdU Cell Proliferation Assay Kit to perform Cell cytotoxicity / Proliferation assay cell type - BxPc-3 human primary pancreatic adenocarcinoma
Get tips on using CD273 (PD-L2) Antibody, anti-mouse, PerCP-Vio® 700 to perform Flow cytometry Anti-bodies Mouse - CD273/PD-L2
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 Monoclonal Anti-γ-Tubulin antibody produced in mouse to perform Western blotting tubulin gamma
Get tips on using Monoclonal Anti-Caveolin-1 antibody produced in mouse to perform Western blotting Caveolin-1
Get tips on using PI/RNase Staining Buffer to perform Cell cycle assay mouse - C2C12
Get tips on using PI/RNase Staining Buffer to perform Cell cycle assay mouse - L929
Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.
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