To control for potential interactions between the TRAg-coated Nile Red beads and the anti-CD71 beads used to enrich for reticulocytes, the assay was also perfomed with reticulocytes enriched using Percoll alone. Both PVP01_0000100 and the positive control PvRBP2b bound to Percoll-enriched reticul...
To compare the anti-amyloid activity, such as inhibition of Aβ42 oligomer and fibril formation by Cur or SLCP, the dot blot assay was used [14]. Briefly, Aβ42 peptide was dissolved in HFIP, vortexed for 1 min and allowed to solubilize for 30 min at room temperature. Then, the HFIP...
The successful generation of oligomeric ADDLs was verified by electron microscopy and a dot blot using the oligomer-specific antibody A11 (Additional file 1: Figure S2a and b). The generated ADDLs bound to mHippoE-14 cells (Additional file 1: Figure S2c). An MTT assay further- more ...
Worm pellets were lysed by sonication and the protein concentration of the lysate was determined using a Pierce BCA protein assay kit (Thermo Scientific, 23227). The fatty acid C13:0 (NuChek Prep; dissolved in methanol) was added to each sample to serve as the internal reference control for ...
Assuming that this may lead to elimination of free AA as a substrate for 15LO, we determined binding of recombinant purified PEBP1 with phospholipids and AA using a dot-blot assay and found that only AA demonstrated marked binding (Figure 6C). Furthermore, on native PAGE gels, free AA ...
After protein concentrations were determined using Pierce BCA Protein Assay Kit (Thermo Fisher Scientific), cell lysates were mixed with Laemmli sample buffer and heated for 10 min at 75 °C before SDS–PAGE. For immunoprecipitation in Fig. 2d, cells were lysed in 500 μl of lysis ...
These cells were subsequently examined for their cholesterol content using a sensitive cholesterol oxidase-based assay using the Amplex Red cholesterol kit (Molecular Probes, Eugene, OR). Statistics Analysis Significant statistical differences between groups were conducted using Student's t test and ...
An MTT assay analysis indicated that after 24 h, cells treated with DFO or Dp44mT showed a slight decrease in cell proliferation. After Dp44mT treatment for 48 h, cell proliferation was reduced by 69%, whereas DFO treatment elicited a decrease in cell proliferation by 45%. After 72 h, ...
HeLa cells were transfected with or without ATG14 siRNA for 48 h and analyzed via western blot. For imaging assay, LDs were labeled with BODIPY-493/503. The nuclei were stained with DAPI. Number of LDs in each cell was counted from 50 cells of three independent experiments.j,kHeLa ...
In contrast to injury-driven lymphangiogenesis, which is promoted by the release of VEGF-C by inflammatory cells26, in this micropocket assay an inflammatory response is not a prerequisite for neovascularization27 and can therefore be used to determine the direct effect of VEGF-A on ...