The use of biological buffers such as TRIS-HCl and TRIS in cell therapy
Release time:
2022-04-19
During cell therapy, biological buffers are sometimes used to separate cells for culture. Now, we will introduce three biological buffers used in cell therapy: TRIS-HCL, TRIS, and CHAPS.
For ester blot analysis: lyse brain tissue and co-cultured cells in ice-cold lysis buffer (20 mM Tris–HCl, pH 7.5, 1 mM EDTA, 5 mM MgCl, 1 mM dithiothreitol, 0.1 mM benzyl) Sulfonyl fluoride plus protease inhibitor cocktail. Lysates were centrifuged.
Preparation of fluorescently labeled FCS: The sample was then added to 0.5 g of FITC dissolved in 5 ml of DMSO. After shaking vigorously for 10 min, the reaction was incubated at 4°C for 16 h and then stopped by the addition of 0.1 volume of 1 M Tris-HCl buffer (pH 8.0).
For tumor immunotherapy: First add manganese chloride (MnCl2) solution to OVA solution and incubate for 20 min with stirring. The DAP solution was dropped into the above solution, and the pH of the mixture was adjusted to pH 7.8-8 by tris(hydroxymethyl)aminomethane-hydrogen chloride (Tris-HCl) buffer.
For the treatment of hepatoma cells: Enzyme assay with artificial substrate For MMP-1 (collagenase-1) assay, the enzymatic activity was measured in reaction buffer [300 mM NaCl, 10 mM CaCl2, 0.005% Brij35, 0.01% NaN3 and 50 mM Tris -HCl (pH 7.5)] using 20 μM MOAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2 as substrate (incubate at 25°C for 90 minutes).
Preparation for antigen-targeted cancer immunotherapy, recombinant large HSP, and tumor protein antigens in lysis buffer, binding buffer, wash buffer, and elution buffer.
To examine the role of exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs) and NPCs in their interactions with the corresponding cells. BM-MSCs, NPCs and Exosomes, EDTA, sodium orthovanadate, sodium fluoride, leupeptin, and 1 mM PMSF.
To assess stem cell DNA integrity for cardiac cell therapy, for cell lysis buffer, dissolve 100 mL of lysis buffer (2.5 M NaCl, 0.1 M EDTA, 10 mM Tris, 1% Triton X-100) in deionized water ( DIH2O). Then, 10 M NaOH was added to adjust the pH to 10.0. Cool the buffer to 4°C.
For Parkinson's disease, PVDF membranes were washed with buffer (0.05% Triton and 0.9% NaCl in 50 mM Tris, pH 7.3), immersed in blocking solution for 60 minutes, and then reacted with secondary antibodies for 1 hour at room temperature.
CHAPS:
For separation of HLA ligands. HLA class I molecules were isolated using standard immunoaffinity purification. Lyse snap-frozen GSC pellets or snap-frozen primary glioblastoma tissue samples in 10 mM CHAPS/PBS containing 1× protease inhibitors.
For potent apo-IDO1 inhibitor cancer immunotherapy. Recombinant IDO1 activity assay Recombinant IDO1 activity was measured at 25°C or 37°C as previously described: 100 nM holo-IDO1 in 100 mM potassium phosphate buffer (pH 7.2) and 1 mM CHAPS supplemented with 0.5% (v:v ) Catalase (25 µl) was added to the inhibitor in DMSO vehicle (0.5 µl DMSO volume) and incubated at the indicated time and temperature (variable).
Hubei Xindesheng is a manufacturer engaged in the production of biological buffers. At present, it produces many types of biological buffers, including TRIS buffer, TRIS-HCL buffer, CHAPS buffer, etc. It is a manufacturer integrating R&D and production. , If you have purchase needs, please contact us!
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