Flowcytometry TLR4 (CD284)

Get tips on using Anti-Human CD284 (TLR4) to perform Flowcytometry TLR4 (CD284) - Mouse / IgG1, kappa Human Brilliant violet 421

Get tips on using TLR4 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - Neuro 2a TLR4

Get tips on using Tlr4 Mouse siRNA Oligo Duplex to perform siRNA / miRNA gene silencing Mouse - BV2 TLR4

Get tips on using Anti-Human CD282 (TLR2) FITC to perform Flowcytometry TLR2 (CD282) - Mouse / IgG1, kappa Human FITC

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.

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

Get tips on using Anti-Human CD3 PE-Cyanine7 to perform Flowcytometry CD3 - Mouse / IgG1, kappa Human PE-Cyanine7

Get tips on using CD14 Antibody, Pacific Orange™ conjugate to perform Flowcytometry CD14 - Mouse / IgG2a Human Pacific Orange™

Get tips on using Alexa Fluor 700-labeled anti-CD16 to perform Flowcytometry CD16 - Mouse /IgG1, kappa Human Alexa Fluor 700

Get tips on using Anti-Human CD56 (NCAM) APC-eFluor® 780 to perform Flowcytometry CD56 (NCAM) - Mouse / IgG1, kappa Human APC-eFluor 780