Site Directed Mutagenesis (SDM) Human Deletion PC-3

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 MYH9 CRISPR/Cas9 KO Plasmid (h) to perform CRISPR Rat - Deletion PC12 myosin IIA (Myh9)

Get tips on using Hs_TET3_2 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - HCT-116 TET3(TET methylcytosine dioxygenase 3)

Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Mouse - Deletion 3T3-L1 SWELL1

Get tips on using Low Input Quick Amp Labeling Kits to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp

Get tips on using Ovation® RNA Amplification System V2 to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp

Get tips on using miRNA Complete Labeling and Hyb Kit to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp

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.

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 pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion AR42J FICD