Get tips on using Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb #2078 to perform Western blotting IKK Alpha
Get tips on using Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb #4806 to perform Western blotting NF-kB
Get tips on using CONFIRM anti-Estrogen Receptor (ER) (SP1) Rabbit Monoclonal Primary Antibody to perform Immunohistochemistry Human - ER
Get tips on using Anti-trimethyl-Histone H3 (Lys4) Antibody, clone MC315, rabbit monoclonal to perform ChIP Anti-bodies H3K4me3
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.
Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.
Get tips on using Anti Type X Collagen (raised against rat) pAb (Rabbit, Antiserum) to perform Immunohistochemistry Mouse - Col X
Get tips on using Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 568 conjugate to perform Flowcytometry Secondary Antibody - Goat Mouse Alexa Fluor 568
Get tips on using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb #9728 to perform ChIP Anti-bodies H3K27me2
Get tips on using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb #5326 to perform ChIP Anti-bodies H3K4me1
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