siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α

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Get tips on using Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade to perform ChIP H3K36Me3 - Sheep Rat -NA-

Products Abcam Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade

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.

RNA RNA isolation / purification Cells immortalized EBL (embryonic lung cell)

Get tips on using Recombinant Anti-SOX9 antibody [EPR14335] (ab185230) to perform Immunohistochemistry Rat - Sox9

Products Abcam Recombinant Anti-SOX9 antibody [EPR14335] (ab185230)

Get tips on using Recombinant Anti-PRMT5 antibody [EPR5772] (ab109451) to perform Immunohistochemistry Rat - PRMT5

Products Abcam Recombinant Anti-PRMT5 antibody [EPR5772] (ab109451)

Get tips on using 17 beta Estradiol ELISA Kit (ab108667) to perform ELISA Rat - Estradiol

Products Abcam 17 beta Estradiol ELISA Kit (ab108667)

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Cellular assays ROS assay cell type human primary corneal epithelial cells

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Proteins ChIP Mouse 3T3-L1 cells

Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Whole genome profiling - rat liver tissue

Products Zymo Research EZ DNA Methylation kit

Get tips on using LC3B Antibody to perform Autophagy assay cell type - Rat cerebral coritcal tissue

Products Novus Biologicals LC3B Antibody

Get tips on using Minimum Essential Medium Eagle to perform Stem cell culture media Rat BMSC

Products Sigma-Aldrich Minimum Essential Medium Eagle

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