Q&A Oct 26, 2025

Organoid Seeding Protocol on Chips MEA Chip Preparation Please refer to the supplier’s protocol for chip surface preparation.For MaxOne single-well chips (Maxwell Biosystems), the preparation includes:Cleaning with 1% Terg-a-zyme solution for a minimum of 2 hours at room temperature.Washing thoroughly with distilled water.Sterilizing with 70% ethanol.Drying under laminar airflow in a sterile cell culture hood.Poly-L-Ornithine Hydrobromide Coating Dissolve Poly-L-Ornithine hydrobromide (Sigma P36...

Q&A Oct 17, 2025

I have experience with intravital imaging using the Zeiss Lightsheet Z.1 with zebrafish embryos and larvae at early developmental stages. In addition, I have also performed intravital imaging of living killifish, using a transparent line as described in the respective publication. For all experiments, I used the standard chamber (there is no extra-large chamber available, and it would not fit into the setup anyway), which means that the samples need to be small enough to fit into the chamber. Th...

Q&A Oct 13, 2025

 AbstractThe ABCC11 gene, through a single nucleotide polymorphism (SNP; rs17822931), is established as a primary determinant of human axillary secretion. Such a composition dictates the presence of both a binary phenotype of earwax (dry/wet) and the absence or presence thereof of characteristic body odor. However, significant phenotypic heterogeneity exists within genotypic groups, suggesting a more complex genetic architecture. This paper proposes a functional genomic model that expands upon t...

Q&A Sep 20, 2025

Abstract Divalent metal ion transporters are conserved across all domains of life and play essential roles in diverse processes such as manganese acquisition during nutritional immunity in bacteria [1] and iron homeostasis in higher eukaryotes [2], [3]. Traditional techniques, such as electrophysiological assays, are often unsuitable due to the slow kinetics of many membrane transporters. To overcome these limitations and to investigate both the activity and ion selectivity of transporters, also...

Q&A Sep 18, 2025

If tissue detachment occurs during 4i we recommend the following:Pipette very gently. If the tissue does not show signs of detaching after the first iteration of 4i but later comes off, likely the problem is pipetting too forcefully and not with the mounting. This is the most common cause of tissue detaching.Dehydrate the tissue longer. After mounting and removing the PBS the tissue can be left to dry out completely (1hour or even overnight). While this is quite effective, it can mask some epito...

Q&A Sep 12, 2025

 Load the needle with a few uls of whatever you’re injecting.For fluorescent tracers, I usually start with 5 mg/ml concentration of tracer and add my phenol red as I would for single cell injections to see where things goPlace the larvae upside down in agarose injection molds (we now use these 3D printed molds: thingiverse.com/thing:4968318), with their hearts facing up. I like to have the higher edge of the mold on the opposite side of where there needle is coming from.Gently pierce the cardiac...

Q&A Sep 11, 2025

We perform PCA using all backbone atom (N,O,C,CA) of the full degradation machinery complex using Cpptraj. Input file is attached below. Then using Normal Model Wizard (NMwiz) plugin in VMD to visualize and observe the essential motion. Specifically, PCA allow us to observe motion by moving your protein along the direction of eigenvector in each eigenmodes.   Below are input file of performing PCA: parm protein.prmtop                           # load your prmtop heretrajin dBET1_r2_400ns_nowat.d...

Q&A Sep 10, 2025

Alexandra Graninger1, 2  and Prasanna Satpute-Krishnan2, * 1Neuroscience Program, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.2Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.*For correspondence: prasanna.satpute@gmail.com AbstractThe antibody-uptake assay is a commonly used technique to monitor endocytosis of integral membrane proteins including transmembrane and glycosylphosphatidylinositol-an...

Q&A Sep 9, 2025

SummaryThis protocol outlines the bioinformatics workflow for hierarchical clustering of H3K27ac peaks. Software and datasetsbwa (https://bio-bwa.sourceforge.net/)samtools (https://www.htslib.org/)bedtools (https://bedtools.readthedocs.io/en/latest/)Picard (https://broadinstitute.github.io/picard/)MACS (https://github.com/macs3-project/MACS)fastp (https://github.com/OpenGene/fastp)fastqc (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)UCSC tools, bedgraphTobigWig (https://hgdownload....

Q&A Aug 26, 2025

Liposome reconstitution of SLC26A6 Materials:1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), (Avanti Polar Lipids)1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG), (Avanti Polar Lipids)Triton X-100 (Sigma)Biobeads SM-2 adsorbents (BioRad Laboratories)Polycarbonate Membranes 400 nm (Sigma)Diethyl ether (Sigma)Avestin Extruder kit (Sigma)      Cary 60 UV-Vis Spectrophotometer (Aligent) Lipid Extraction:Pool desired Lipids in Glass-flask at a ratio of 3:1 POPE:PO...