*Washing Buffer Recipe: To make the citrate-phosphate buffer, pH 5, mix 4.7 g citric acid (MW=192) with 9.2 g dibasic sodium phosphate (Na2HPO4) dehydrate (MW=178). Fill up to 1 L with distilled w...
However, if the breaking buffer is used for S. cerevisiae, the EDTA should be omitted. In other words, if working with Pichia, the breaking buffer should contain ETDA (see Page 59 of the Pichia manual for ...
Empty CellSolutionRecipe A.For sucrose cushion; 6% sucrose, 0.06% TX-114 in PBS 10 ml PBS 0.6 g sucrose 50 μl 11.4% TX-114 (see below) B.0.5% TX-114 in PBS 10 ml PBS 500 μl 11.4% TX-114 (see below) C.Pre-condensed 11.4% TX-114 in PBSSee text ...
Low affinity or transient, and may be lost during the stringent washing procedures with lysis buffer. Although spectral counts have a correlation with protein abundance other methods, for example, SILAC offer more precise strategies for quantitative proteomics53. Nevertheless, by mapping the IAV–host...
Immunoblotting analysis was performed following the procedure outlined in our previous study [34]. To perform immunoprecipitation, add the primary antibody to the cell lysate and rock overnight at 4 °C. Protein A beads were thoroughly washed with lysis buffer, and the supernatant was analyzed ...
To make sure the proteins are fit and ready and in sufficient quantities, your extraction technique requires a fine balance of reagents, temperatures and procedures. Formulate your recipe by choosing from decades-old physical methods as well as newer detergent- and enzyme-based approaches. ...
Recombinant proteins and cell extracts were incubated 1 hour and 30 minutes at 4°C, and washed four times in lysis buffer. In vitro binding assay To remove contaminant bacterial nucleic acids, recombinant proteins were pretreated with nucleases (0.25 U/μl DNase I and 0.2 μg/μl RNase) ...
(PBS). Total cell lysates were obtained by lysing the oocytes in Pierce™ IP lysis buffer with added Halt protease inhibitor cocktail (Thermo Fisher Scientific). Surface proteins were purified with the Pierce™ Cell Surface Protein Isolation Kit (Thermo Fisher Scientific). Purified surface ...
(0.5 M in Tris pH 8.0 0.1 mM) and twice with lysis buffer, boiled with Laemmli sample buffer, and then were subjected to western blot analysis. Antibodies used for western blot analysis and immunoprecipitations were: α-FLAG antibody purchased from Sigma-Aldrich Co. (F1804) with 1:...
Recipe 1: HEN Buffer1 Hepes-NaOH pH 7.7250mM EDTA1mM Neocuproine (Sigma)0.1mM Recipe 2: CHAPS Stock Prepare a 10% solution in dH2O. Recipe 3: MMTS Stock Prepare a 2-M solution in dimethylformamide. Recipe 4: Blocking Buffer2 HEN buffer9 volumes ...