crispr-mouse-activation-3t3-l1-c-ebp

Get tips on using Cell Comb™ Scratch Assay to perform Wound healing assay cell type - mouse 3T3-L1

Get tips on using pMSCV-LTR-dCas9-VP64-BFP to perform CRISPR Mouse - Activation Neuro-2a Sim1

Get tips on using DeadEnd™ Colorimetric TUNEL System to perform TUNEL assay cell type - 3T3 L1 mouse adipose tissue

Get tips on using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 to perform ChIP Mouse - 3T3-L1 cells

Get tips on using QuikChange Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Mouse - 3T3-L1 Clk1

Get tips on using Silencer® Select- Gdf10 siRNA to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 BMP-3b/GDF10

Get tips on using Mouse CRP / C Reactive Protein / PTX1 PicoKine™ ELISA Kit to perform ELISA Mouse - 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.

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).

Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - 3T3 L1 mouse adipose tissue