Q&A Apr 30, 2026

Proteomic profiling of hypusinated protein candidates using a clickable spermidine probe Author List: Tian Zhang1,2,4, Jianlong Li1,2,4, Xinbo Hu1,4, Zhaoyin Wang1, Yaoyang Zhang1,3, Junying Yuan1,3,6*, Bing Shan1,3,5* Affiliations:1Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China;2University of Chinese Academy of Sciences, Beijing, China;3Shanghai Key Laboratory of Aging Studies, Shanghai, China;4Th...

Q&A Apr 29, 2026

ABSTRACT:Quantitative assessment of cartilage architecture is essential for understanding skeletal development, tissue homeostasis, and disease progression; however, standardized and accessible workflows for reproducible morphometric analysis remain limited. This paper outlines a reproducible protocol using FIJI (Fiji Is Just ImageJ) software to quantitatively measure cartilage properties from histological images of stained tissue sections.The protocol enables the measurement of key parameters, ...

Q&A Apr 27, 2026

Key featuresProvides an efficient protocol for CT infection in McCoy B cells, followed by lugol stain.Offers a low-cost option, in comparison to fluorophore-conjugated secondary antibodies. KeywordsChlamydia trachomatis; McCoy B cells, Cell culture; Lugol iodine staining AbstractChlamydia trachomatis (CT) is an obligate intracellular bacterium that requires growth inside a mammalian host cell for propagation and survival. CT is the most common etiological agent of bacterial sexually transmitted ...

Q&A Apr 26, 2026

Intrathecal procedureCapture the monkey at least one week before the treatment. The monkeys will be individually homed and enrichment toys will be provided. Fast the monkey at least 12 h before the procedure (will begin in the morning). Sedate the monkey with 6.7 mg/kg ketamine (intramuscular) and allow it to fall into deep sleep (at least 15 minutes). Clean the area around the injection site with 70 % ethanol before administering 0.03 mg/kg buprenorphine hydrochloride (intramuscular) for pain m...

Q&A Apr 22, 2026

Protein extraction1. HeLa cells are grown in DMEM in 6 cm dishes at 37°C in a 5% CO2 incubator. 2. Remove DMEM. 3. Add 1 mL of ice-cold PBS. 4. The cells are harvested using a scraper and centrifuged at 2,000 × g for 2 min at 4°C. 5. Remove supernatants. 6. Freeze cell pellets once at -30°C or -80°C. 7. Solubilize the cell pellets with 120 uL of ice-cold Lysis Buffer (containing 0.2% DDM) for 20 minutes on ice. 8. Divide the cell suspension into 90 uL and the remainder (~30 uL). 9-1. Centrifuge ...

Q&A Apr 21, 2026

DPPH Radical Scavenging Assay for Evaluation of Antioxidant Activity in Plant ExtractsAbstractThis protocol describes the determination of antioxidant activity using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay. The method is based on the reduction of DPPH radicals by antioxidants, resulting in a color change from purple to yellow, which is measured spectrophotometrically at 517 nm. The antioxidant capacity is expressed as percentage inhibition and IC₅₀ values.Materials and ...

Q&A Apr 21, 2026

Detailed Protocol: PBMC Isolation, Culture, and RNA ExtractionI. PBMC IsolationCollect peripheral blood samples in EDTA tubes. Gently invert the tubes several times to ensure proper mixing. Dilute blood 1:2 with HEPES-containing buffer. For each 9 mL blood tube, bring the final volume to 25 mL in a 50 mL Falcon tube using buffer. Add 12.5 mL Ficoll to a new 50 mL Falcon tube. Carefully layer the 25 mL diluted blood over the Ficoll. Centrifuge at 2300 rpm for 20 min, with acceleration and brake s...

Q&A Apr 17, 2026

We used an immortalized human umbilical vein endothelial cell (HUVEC) line. In the published Methods, these cells were maintained in RPMI medium supplemented with 10% fetal bovine serum (FBS), streptomycin, and penicillin, at 37 °C in 5% CO₂.For the migration and adhesion experiments in the paper, cells were first grown in complete RPMI and then switched to RPMI containing 0.2% BSA during the assay phase. For migration assays, HUVECs were seeded at 5 × 10^5 cells/well in 6-well plates. After 2 d...

Q&A Apr 15, 2026

Detailed protocol for Johnsen scoring in testicular histopathology used in our study  BackgroundThe Johnsen scoring system is a semiquantitative histopathological method used to evaluate spermatogenesis in seminiferous tubules. It assigns a score from 1 to 10 according to the most advanced germ cell type present and the degree of spermatogenic maturation. Higher scores indicate more advanced spermatogenesis, and a score of 8 or higher generally indicates the presence of spermatozoa.SpecimensTest...

Q&A Apr 14, 2026

AbstractVoltage-gated ion channels such as hERG (KV11.1) adopt multiple conformational states that are critical for ion conduction and drug binding, yet experimentally resolved structures often capture only a limited subset of these states. Here, we describe a protocol for generating hERG channel models in closed, open, and inactivated conformations using ColabFold AlphaFold2 notebook with template-guided sampling (1). The approach uses carefully constructed structural templates derived from hom...