{"id":480,"date":"2018-06-26T17:41:43","date_gmt":"2018-06-26T21:41:43","guid":{"rendered":"https:\/\/www.lifetein.com\/blog\/?p=480"},"modified":"2024-11-16T22:02:36","modified_gmt":"2024-11-17T03:02:36","slug":"how-to-measure-the-cellular-uptake-of-peptides","status":"publish","type":"post","link":"https:\/\/www.lifetein.com\/blog\/how-to-measure-the-cellular-uptake-of-peptides\/","title":{"rendered":"How to measure the cellular uptake of peptides?"},"content":{"rendered":"

TMR\/Cy3\/Cy5 was introduced for the fluorescent label with peptides to evaluate each peptide’s cell-penetrating ability and intracellular distribution.<\/p>\n

    \n
  1. Cells (HeLa or Huh-7) were seeded on 24-well culture plates (40,000 cells\/well) and incubated in 400 \u03bcL of DMEM containing 10% fetal bovine serum (FBS).<\/li>\n
  2. The medium was then replaced with fresh medium containing 10% FBS and Tetramethylrhodamine carboxylic acid (TMR)-labeled peptides.<\/li>\n
  3. The solution was added to each well at an appropriate concentration (e.g., 0.5 uM, 1 uM, 1.5 uM, 2 uM).<\/li>\n
  4. After 1, 2, 3, or 4 hours of incubation, the medium was removed, and cells were washed with ice-cold PBS and trypsin.<\/li>\n
  5. After adding medium containing 10% FBS, cells were centrifuged at 1600 rpm for 3 min at 4 \u00b0C. The cell pellets obtained were suspended in ice-cold PBS, centrifuged at 1600 rpm for 3 min at 4 \u00b0C, and then treated with Cell lysis buffer.<\/li>\n
  6. The fluorescence intensity of each lysate was measured using a spectrofluorometer. The amount of protein in each well was concomitantly determined using the BCA protein assay.<\/li>\n
  7. The results are presented as the mean and standard deviation from 3 samples.<\/li>\n<\/ol>\n

    Reference: https:\/\/www.nature.com\/articles\/srep19913#s1<\/a><\/p>\n

    \"Cy5

    Cy5 labeled peptide in cells<\/p><\/div>\n","protected":false},"excerpt":{"rendered":"

    TMR\/Cy3\/Cy5 was introduced for the fluorescent label with peptides to evaluate each peptide’s cell-penetrating ability and intracellular distribution. 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