Cells were washed with HBSS + 5% FBS and Prominin1-positive cells were sorted directly in cold RLT lysis buffer (Qiagen) + 1% 2-mercaptoethanol. Unstained cells and isotype-specific IgG-FITC labeled cells served as negative control for gating. Isolated RNA was sequenced at the iGE3 ...
To estimate ER steady-state [Cl–], the cells were suspended in HBSS buffer containing 2 mM Ca2+ and 140 mM Cl–. To characterize the [K+] sensor in vitro, the K+ binding protein (mNG-Ec-Kbp-E12A), also named KRaION2 by a previous report,28 was fused with monomeric DsRed ...
Trypsin (1:250), powder (Cat. No. 27250018) is prepared by adding the 0.25 g of Trypsin powder to 100 mL of chilled HBSS (Cat. No. 14170112) to create a 0.25% solution. For 1% solution, you would add 1g of trypsin powder to 100 mL of chilled HBSS. ...
[6]. After thawing, cells were washed with Hanks' balanced salt solution (HBSS) and further processed for RNA isolation. AML samples treated according to this procedures usually contain greater than 90% blasts after thawing [6]. A total of 162 de novo AML patients who had been referred to...
THP-1 WT/OE cells w/o Tunicamycin (TM, Yeasen 60251ES03) treatment were loaded with Rhod-2 in HBSS buffer supplemented with 200 nm ER-Tracker Blue-White DPX for 30 min at 37 °C. Cells were then fixed with 4% formaldehyde at 37 °C for 10–20 min, washed with phosphate buffered ...
Several long noncoding RNAs (lncRNAs) have been shown to function as components of molecular machines that play fundamental roles in biology. While the number of annotated lncRNAs in mammalian genomes has greatly expanded, studying lncRNA function has be
26、 ofthChildren HbssitalwillcarefortheseriousIRinjuredpupils.4) severalph on ecallsmak inginq uiriesaboutthepositi ono ftheChiefF inan cialOfficer.5) straighte no utallRourfi nan cialproblemsifRoujoi no urclub.3.1)i nq uirR;diedofh un ger;peoplesurvied2)l nsta ntlR;giveuphis;retire...
Cells were washed three times using HBSS and suspended in neuronal media: Neurobasal media plus 2% B27, 1 × glutamine, and 1 × pen/strep. Analysis of rat cortical neurons were performed to confirm identity using immunofluorescence of Tju1 (Sigma, 1 : 1000), Satb2 (Abcam, 1 :...
Nonadherent cells were removed by gentle washing three times with HBSS. Adherent cells were cultured in DMEM containing 10% FBS, 10 mM HEPES, 100 U/mL penicillin, and 100 µg/mL streptomycin for 24 h before experi- ments. Kupffer cells were plated at a density of 5 × 105/well in ...
. After 3 hours, the HBSS medium was replaced with Schneider's medium without FBS. Observation and quantification of GFP fluorescence The transfected cells attached on the cover glasses were fixed with 2% final concentration of paraformaldehyde solution for 10 min and washed three times with PBS ...