siRNA / miRNA gene silencing Human hES cell line H1 (WA01)

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Human/Mouse NLRP3/NALP3 Antibody to perform Western blotting NLRP3

Get tips on using Human/Mouse/Rat SOX2 Antibody to perform Western blotting SOX2

Get tips on using Human Ubiquitin/Ubiquitin+1 Antibody to perform Western blotting Ubiquitin

Get tips on using Human BDNF ELISA Kit (ab212166) to perform ELISA Mouse - BDNF

Get tips on using Human/Mouse BDNF DuoSet ELISA to perform ELISA Mouse - BDNF

Get tips on using Human SCF ELISA Kit (ab100636) to perform ELISA Rat - SC

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.

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Get tips on using Human VWF / von Willebrand Factor ELISA Kit to perform ELISA Human - VWF-A2