Get tips on using Purified Rat Anti-Mouse IFN-γ to perform Flow cytometry Anti-bodies Mouse - IFN-γ
Get tips on using APC Rat Anti-Mouse IFN-γ to perform Flow cytometry Anti-bodies Mouse - IFN-γ
Get tips on using Purified Rat Anti-Mouse Siglec-F to perform Flow cytometry Anti-bodies Mouse - Siglec F
Get tips on using BV421 Rat Anti-Mouse Siglec-F to perform Flow cytometry Anti-bodies Mouse - Siglec F
Get tips on using PE anti-mouse/rat CD29 Antibody to perform Flow cytometry Anti-bodies Mouse - CD29/β1-Integrin
Get tips on using ScriptSeq Complete Kit (Human/Mouse/Rat) to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)
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/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA to perform ELISA Rat - HSP70
ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.
Get tips on using Purified anti-mouse/rat/human FOXP3 Antibody to perform Western blotting FOXP3
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