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Site Directed Mutagenesis (SDM) Human

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Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Products STEMCELL technologies MammoCult™ Human Medium Kit

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres

Products STEMCELL technologies MammoCult™ Human Medium Kit

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.

Cellular assays TUNEL assay cell type HNSCC Detroit 562 human head and neck tumor cells

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

RNA RNA isolation / purification Tissue Human Seminal vesicles

Get tips on using p62 (human) polyclonal antibody to perform Immunohistochemistry Mouse - p62

Products Enzo Life Sciences p62 (human) polyclonal antibody

Get tips on using Human MMP-1 Antibody to perform Western blotting MMP1

Products R&D Systems Human MMP-1 Antibody

Get tips on using Human GDNF DuoSet ELISA to perform ELISA Rat - GDNF

Products R&D Systems Human GDNF DuoSet ELISA

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 primary human pancreatic stellate cells

Get tips on using Human VEGF PicoKine™ ELISA Kit to perform ELISA Human - VEGF

Products BosterBio Human VEGF PicoKine™ ELISA Kit

Get tips on using Human Sonic Hedgehog ELISA Kit (ab100639) to perform ELISA Human - ShhN

Products Abcam Human Sonic Hedgehog ELISA Kit (ab100639)

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