Site Directed Mutagenesis (SDM) Rat Point mutation Rat-2

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #13082 to perform Autophagy assay cell type - RAW 264.7

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

Get tips on using ON-TARGETplus Human PLK1 (5347) siRNA - Individual to perform siRNA / miRNA gene silencing Human - MIA PaCa-2 PLK-1

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product is observed in the experimental sample, add more DNA to the PCR reaction or increase the number of amplification cycles. Furthermore, if you have any problem with antibodies, make sure to use the ChIP-validated antibody.

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 DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - SK-MEL-2 human melanoma

Get tips on using Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - Sao2-2, MG-62

Get tips on using CyQUANT® Cell Proliferation Assay Kit to perform Cell cytotoxicity / Proliferation assay cell type - Saos-2

Get tips on using QIAamp 96 Virus QIAcube HT Kit (5) to perform RNA isolation / purification Viral - SARS-CoV-2

Get tips on using VWR Life Science RIPA Lysis Buffer, Biotechnology Grade to perform Protein isolation Mammalian cells - Caco-2