Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Cells - Immortalized cell lines HeLa
Get tips on using ApopTag® Peroxidase In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - FaDu human squamous cell carcinoma
Get tips on using BrdU Cell Proliferation Assay to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma
Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma
Get tips on using Cell Proliferation ELISA, BrdU to perform Cell cytotoxicity / Proliferation assay cell type - MDA-MB-231 breast adenocarcenoma
Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - LTEP-a-2 lung adenocarcenoma
Get tips on using Cell Counting Kit-8 to perform Cell cytotoxicity / Proliferation assay cell type - SMMC-7721, Huh7, Hep3B, 293T
Get tips on using Purified Mouse Anti-Human ZO-1 Clone 1/ZO-1 (RUO) to perform Western blotting ZO-1
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.
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.
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