Rabbit anti-b-tubulin antibody (#2146) was purchased from CST Japan K.K. (Chiba, Japan). The concentration used in immunofluorescence staining and Western blot analysis was in accordance with the recommended protocol for each antibody. Oil-red O staining Cultured cells were washed with PBS and...
Apo(a) in normal human serum can be detected at normal size with the same monoclonal antibody. Moreover, purchased monoclonal antibody to Lp(a) had been prepared to be highly specific to human apo(a) and to be not reactive to plasminogen. According to our results, it was suggested that ...
Transplacentally acquired maternal antibody can provide a short-term protective umbrella, so such examples tend to be more severe if the mother is undergoing a primary infection and the infant is born without maternal antibodies. The tendency for very young infants with respiratory syncytial virus ...
If utilizing alternate cell type, excessive primary/secondary antibody or dye concentrations for cell type – titrate dilutions to optimize signal. Additional wash steps after the primary/secondary antibody incubations may help decrease background fluorescence due to non- specific binding. Check for ...
(H, I) Immunofluorescence assay for mitochondria using a mitochondrial-specific antibody Tom-20. The average length of the mitochondria was measured, and this parameter was used to quantify mitochondrial fission. (J, K) The expression of MIEF1 was also evaluated via Western blo...
They were then washed with TBS-T and incubated with the corresponding peroxidase-conjugated secondary antibody (anti-rabbit, anti-goat or anti-mouse) used at 1:8000 dilution. The chemiluminescent signal induced by ECL reagent (GE Healthcare) was detected in a ChemiDoc image analyzer (Bio-Rad)...
For cytoskeletal proteins, either a rabbit anti-b-tubulin (diluted 1:10 in 3% BSA in PBS and revealed by a Texas red-conjugated anti-rabbit-IgG antibody) for 1 h or a 10 mM FITC-conjugated phalloidin for 20 min were used. All the incubations were performed at room tem- perature. ...
Assay of Reactive Oxygen Species (ROS) Generation The DCFH-DA method was used to determined the effects of 1–3 on ROS generation as reported68. In this method, the stable non-fluorescent DCFH-DA was hydrolyzed by esterases in cells to non-fluorescent DCFH, which was rapidly oxidized by ROS...
Moreover, unlike the previous study [45], where the antibody used was only capable of measuring TG2-L expression, the antibody used in the current study proved to be cross-reactive to both TG2-L and TG2-S isoforms, confirming that TG2-S levels were also unchanged during the development...
Figure 1. HepG2 cells’ viability upon 24 h exposure to paraquat (PQ) (A) and to Ginkgo biloba leaf extract (B). Cell viability was assessed using Alamar Blue assay (see Methods), data are expressed as % of control (Cont.; non-treated cells) and are shown as mean ± S.D (n =...