FLUORESCENCEPROTEINSSIGNAL-to-noise ratioWESTERN immunoblottingUBIQUITIN ligasesRecombinant proteins in complex solutions are typically detected with tag-specific antibodies in Western blots. Here we describe an antibody-free alternative in which tagged proteins are detected directly in polyacrylamide gels. For...
A tunable broadband Ti:sapphire laser (Chameleon, Coherent) was used at 780 nm to capture Alexa Fluor 488 and 546 fluorescence. For each mouse, four to eight z-stacks with a step size of 0.51 µm were recorded from different positions in the bone marrow. Proplatelet-forming MKs ...
Fluorescence of the SYPRO Orange protein gel stain (S6650, S6651; Protein Detection on Gels, Blots and Arrays—Section 9.3) has been used to follow isothermal protein denaturation (Monitoring Protein-Folding Processes with Environ...
Fluorescence detection multiplexing achieves more data from your sample with iBright. Multiplexing can capture up to four proteins in a single blot for more meaningful and representative experiments, a must for western blot publication. For today’s top journals, total protein normalization and high-qu...
RNA fluorescence in situ hybridization (FISH) enables investigation of gene expression in the context of tissue type. Despite development of FISH methods for plants, easy and reliable whole mount FISH protocols have not yet been reported. Results We adapt a 3-day whole mount RNA-FISH method for...
Fluorescence-activated cell sorting (FACS) We analyzed the infiltration of immune cell subsets in CT26 transplanted tumor tissue by fluorescence-activated cell sorting (FACS) analysis. After the mice were euthanized, the tumor tissues were stripped and cut into pieces, then digested at 37 °C for...
We study the structure and the time evolution of transient gels formed in colloid-polymer mixtures, by means of fluorescence Confocal Scanning Laser Microscopy (CSLM). This technique is used in conjunction with novel colloidal silica particles containing a fluorescent core. The confocal micrographs rev...
The goal of skin resurfacing is the rejuvenation of the skin by stimulation of its regenerative potential. The outer layer of epidermis is ablated in order to improve the quality of the skin. Sun damage is removed and age-related changes are improved in skin resurfacing. In general, there ar...
After two washes in washing buffer (PBS containing Triton 0.4%), secondary fluorescence-conjugated antibodies were added in a dark humid box and incubated for 1 h at room temperature. Slides were washed twice with washing buffer and nuclei were stained with NucBlue Live ReadyProbe Reagent ...
the protease concentration range was 1–5 nM. We monitored the increase of fluorescence intensity with a Tecan infinite 2000 Pro plate reader (Tecan, Switzerland) in continuous mode (excitation at 370 nm, emission at 460 nm) and calculated the corresponding kcat/KMvalues as earlier describ...