We found minimal change in the effect estimate when we added the diagnoses of chronic conditions to the model or when we included the road buffer traffic marker (Table 3).
Cells were then washed in FACScan buffer, and surface marker detection of CD8 and intracellular cytokine staining for IFN-γ were performed as described [ 31].
Western blotting kits, sample buffer, molecular weight marker, nitrocellulose membrane, and filter paper for Western blotting were obtained from Bio-Rad LaBathtories (Corston, Bath, UK).
Protein samples dissolved in the sample buffer and the marker protein were denatured at 100 °C for 5 min before being loaded onto the gel.
When mitochondrial swelling was induced by incubating the mitochondria in hypoosmotic buffer, immunoreactivities of marker proteins of the mitochondrial intermembrane space (adenylate kinase 2 (AK2), cytochrome c) were reduced, whereas STAT3 signal intensity was unchanged (Fig. 3c).
The markers of buffer were indicated following: circles, acetate; squares, citrate; diamonds, phosphate, isosceles triangles, borate-NaOH and right triangles, bicarbonate-NaOH.
Then, 2 μl of the colloidal gold markers in buffer were added and samples blotted and vitrified as for the cellular specimens.
Cells were incubated for 30 min in PBS supplemented with 2% FCS and 0.05% NaN3 (FACS buffer) with antibodies against markers for immune cell subpopulations and for lymphocyte activation.
New England Biolabs supplied all the enzymes and their buffers, protein and DNA markers, plasmids and competent cells.
New England Biolabs supplied all the other enzymes and their buffers, protein and DNA markers, plasmids, and competent cells.
