Reactions were stopped by SDS sample buffer addition and analyzed in western blot assays with a His-tag antibody.
Changes in [Ca2+] in the cytoplasmic buffer upon IP3 (7.5 μM) addition in the presence or absence of ruthenium red (3 μM) was measured by fura2 in a fluorometer (Csordás et al., 2006; Roy et al., 2009b).
Unlike in buffer, addition of exogenous H1 or RanBP7 did not have dramatic effects on chromatin area (Figure 2B), possibly due to the presence of the embryonic linker histone H1M which binds chromatin tightly and does not interact strongly with RanBP7, as well as myriad other chromatin proteins present in the extract [22].
As expected, in cells that were AA-starved in KRB buffer, addition of L-leucine or L-isoleucine, but not L-glutamine, was sufficient to restore mTOR localization to LAMP2+ LE/Ly vesicles in uninfected cells (Fig. 5A).
However, EDTA (2 mM) addition in the buffer could not substitute the tartaric acid precipitation and the AI significantly increased when a desalting column was used for buffer exchange as an alternative to the time-consuming, two-step dialysis.
Several reverse crosslinking temperatures (90 °C, 80 °C, 65 °C, and 56 °C), incubation times (1 and 3 h), and buffer addition (saline sodium citrate) were studied in 33 samples from 5 patients (Supplementary Table S1) using two DNA extraction kits (Supplementary Table S2).
Intake of sodium bicarbonate was higher (P<0.005) when provided free access than when incorporated into the protein pellet (13.9 g/d vs. 5.0 g/d for BF and BI) but it did not affect dry matter intake (797 g/d) or average daily gain (328 g/d vs. 289 g/d for male and female lambs), although barley grain intake tended (P=0.052) to decrease in response to buffer addition.
Mitochondrial light scattering changes 40 sec after the addition in the experimental buffer were used to generate the data shown in the figure.
In hardware, packet loss may happen due to overflow at a finite-depth transmit buffer in addition to the packet corruption in the channel.
We also added mimic miRNA probes with 0, 1, 2 or 3 mismatches to the hybridisation buffer, in addition to the DIG-labeled detection probe, in a competition assay.
