shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y)

Get tips on using Human TNF alpha ELISA Kit (ab181421) to perform ELISA Human - TNF-alpha

Get tips on using Human TNF-alpha Quantikine ELISA Kit to perform ELISA Human - TNF-alpha

Get tips on using Human PDGF BB ELISA Kit (ab100624) to perform ELISA Human - PDGF-BB

Get tips on using Human HO 1 ELISA Kit (ab133064) to perform ELISA Human - HO-1

Get tips on using Human Dkk-1 ELISA Kit (RAB0143) to perform ELISA Human - Dkk-1

Get tips on using Human Dkk-1 DuoSet ELISA (DY1906) to perform ELISA Human - Dkk-1

Get tips on using Human Cytochrome c Quantikine ELISA Kit to perform ELISA Human - Cytochrome C

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using Human ICAM1 ELISA Kit (CD54) (ab100640) to perform ELISA Human - ICAM-1/CD54

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.