CRISPR Rat Deletion INS-1 832/13

Get tips on using BioMag Goat Anti-Rat IgG (500 ml) to perform Cell Isolation Mouse T cells

Get tips on using PU.1 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - RAW264.7 PU.1

Get tips on using p-c-Jun Antibody (KM-1): sc-822 to perform Western blotting C-Jun

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 Purified anti-mouse/rat/human FOXP3 Antibody to perform Western blotting FOXP3

Get tips on using Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit to perform ELISA Mouse - OPN

Get tips on using Rat CRP/C Reactive Protein PicoKine™ ELISA Kit to perform ELISA Rat - C-Reactive Protein/CRP

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 Human/Mouse/Rat/Canine ALCAM/CD166 Antibody to perform Immunohistochemistry Mouse - CD166 / ALCAM

Get tips on using Rat/Mouse Cytochrome c Quantikine ELISA Kit to perform ELISA Mouse - Cytochrome c