where U represents a full buffer set.
To detect intracellular Bcl6, cells were stained in the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) after surface staining.
The pH stability of ScLac was investigated by incubating an aliquot of the purified enzyme preparation in the same buffer set used to determine the pH optima for 0, 1, 6 and 24 h (Fig. 4a).
Endotoxins were removed in an additional step (Endofree buffer set; Qiagen, Santa Clarita, CA).
Intracellular Foxp3 staining was done with Alexa Fluor647- or Alexa Fluor488-labeled anti-Foxp3 using Foxp3 Staining Buffer Set (eBioscience).
After that cells were washed and stained using a permeabilisation buffer (Foxp3 Staining Buffer Set, e-Biosciences).
The Strep purification was done using the Strep-tag® Protein Purification Buffer Set and according to manufactures protocol (IBA).
Intracellular FOXP3 staining was performed using the FOXP3 Fix/Perm Buffer set (Biolegend) according to the manufacturer's recommendations.
For intracellular analysis of Foxp3, cells were fixed/permeabilized using the Foxp3 Staining Buffer Set (eBioscience) according to the manufacturer's instructions.
In some experiments, cells were fixed by a staining buffer set (eBioscience), then intracellularly stained with the antibodies to Foxp3 (PCH101; eBioscience) and Ki67 (BD6; Biosciencesces).
Foxp3 staining was performed, after staining of CD3, CD4 and CD25 surface antigens, using the Foxp3 buffer set from eBioscience according to the manufacturer instructions.
