siRNA / RNAi /miRNA transfection Mouse Primary Splenocytes

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using Monoclonal Mouse Anti-Human Cytokeratin (Concentrate) Clone AE1/AE3 to perform Immunohistochemistry Mouse - Cytokeratin

Get tips on using Purified Rat Anti-Mouse CD24 Clone M1/69 (RUO) to perform Immunohistochemistry Mouse - CD24

Get tips on using Purified Rat Anti-Mouse CD24 Clone M1/69 (RUO) to perform Immunohistochemistry Mouse - CD24

Get tips on using Mouse GFR alpha-3/GDNF R alpha-3 Antibody to perform Immunohistochemistry Mouse - Gfrα3

Get tips on using Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA to perform ELISA Mouse - HSP70

Get tips on using Mouse TIM-1/KIM-1/HAVCR Quantikine ELISA Kit to perform ELISA Mouse - KIM-1

Get tips on using Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit to perform ELISA Mouse - IGF-I

Get tips on using INTERFERin® to perform siRNA / miRNA gene silencing Human - 501 Mel and SK Mel 28 FANCD2 Polymer / Lipid

Flow cytometry is an immunophenotyping technique whereby sing cell suspensions are stained for either cell surface markers or intracellular proteins by fluorescently-labelled antibodies and analyzed with a flow cytometer, where fluorescently-labelled molecules are excited by the laser to emit light at varying wavelengths, which is then detected by the instrument. There are several key criteria which are required to be kept in mind while designing a flow experiment- 1. Antibody titration (optimal dilution of antibodies should be calculated in order to avoid over- or under- saturated signals for proper detection of surface and intracellular markers), 2. Precision (3 or more replicates of the sample should be used per experiment), 3. Specificity (proper isotype controls should be included in the experiment), 4. Day-to-day variability (experiments should be repeated 3 or more times to ensure consistency and avoid variability due to flow cytometer settings), 5. Antibody interaction (Fluorescence minus one or FMO should be used, which is the comparison of signals from panel minus one antibody vs. the full panel), and 6. Antibody stability (fluorescently-labelled antibodies should be stored at 4C).