Additionally, hemolysis results in release of analytes and enzymes from red blood cells (RBCs), which can falsely elevate many test results.5 Destruction of RBCs yields a lower RBC count.1 Excess fluid released from
the lysed RBCs also creates a dilution effect in serum and can result in an artifactually decreased packed cell volume.
WT and Lpar2 - / - T cells were incubated with 1 μM LPA for the periods indicated, and were
lysed immediately thereafter.
After 24 h, cells were incubated with or without PP2,
lysed, and immunoprecipitated with anti-HA (left) or anti-myc (right) or control Abs (ctr Ab) followed by Western blotting with the indicated Abs.
For quantification of phagocytosis, alveolar macrophages were infected at an MOI of 20 for 1 h, then media was removed, extracellular bacteria were killed with gentamycin (200 μg / ml for 10 min at 37 C), and macrophages were
lysed with 0.2 % triton X-100 (10 min at room temperature).
Cells were washed with ice - cold PBS,
lysed for 15 minutes on ice with M - PER protein extraction reagent (Pierce), scraped, and spun at 14,000 rpm for 10 minutes at 4 °C.
Confluent 10 cm plates were fixed with 1 % PFA at room temperature for 10 minutes,
lysed in SDS lysis buffer (50 mM Tris, 10 mM EDTA, 1 % SDS, Roche Complete protease inhibitors, pH 8.1), scraped and collected into 1.5 mL microcentrifuge tubes, and DNA was sonicated to 200 — 800 bp fragments with a Branson Sonifier 250 set to 30 % power / 90 % duty, four 10 second pulses.
Expression of four stem cell marker genes normalized to cyclophilin in nine single cell equivalents prepared from a pool of nine
lysed HES3 cells.
In preclinical research, tumor cells
lysed by coxsackievirus A21 (CVA21), a naturally occurring «common cold» ICAM - 1 — targeted RNA virus, induced a secondary systemic host - generated antitumor immune response.
Cells were washed with PBS three times and
lysed by adding a modified RIPA buffer (50 mM Tris / HCl pH 7.4; 150 mM NaCl, 1 mM EDTA, 1 % Igepal, 0.25 % Na - deoxycholate).
48 hours after transfection, cells were
lysed with 100 µL / well 1 × passive lysis buffer (PLB, Promega) for 15 minutes with shaking.
Antigen - presenting cells then take up
the lysed cells.
Isolation of cells from outer, middle and inner colony zones was performed under microscopic guidance (Figure S4), after which cells were immediately
lysed and prepared for Q - RTPCR.
The cells were
lysed with RIPA buffer.
(A) 106 BaF3 Aut (V658I), Aut (F958V) and BaF3 cells stably transduced with M - RAS or (B) JAK1 V658I and F958V mutants were treated for 30 min with 500 nM CMP6 inhibitor or with 0.1 % DMSO as a control (− condition),
lysed and subjected to Western blot analysis.
(B) BaF3 cells stably transduced with V617F, Y931C or V617F / Y931C mutant were
lysed and subjected to Western blot analysis.
Cultured cells were
lysed in radioimmunoprecipitation assay (RIPA) buffer (50 μl RIPA per 1 × 106 cells; 50 mM Tris / HCl; 150 mM NaCl, 1 % Triton X-100, 1 mM EDTA, 1 mM NaF, 1 mM Na3VO4, 1 μg / ml leupeptin and 1 μg / ml pepstatin) for 1 h on ice, and insoluble debris was removed by centrifugation.
(E) 106 BaF3 cells stably transduced with JAK2 V617F, Y931C and V617F / Y931C double JAK2 mutant were treated for 30 min with 300 nM CMP6 inhibitor or with DMSO as a control (− condition),
lysed and subjected to Western blot analysis.
Cells were
lysed 48 h later with modified radioimmunoprecipitation assay buffer (RIPA buffer, 50 mM Tris - HCl, pH7.4, 150 mM NaCl, 1 % Nonidet P - 40 (NP - 40), 0.25 % sodium deoxycholic acid, 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM phenylmethylsulphonyl fluoride (PMSF), 1X Complete cocktail protease inhibitor (Roche, Penzberg, Germany)-RRB-.
Cells were then
lysed and centrifuged to pellet the nuclei.
For this protocol, samples were
lysed in 1 × RIPK2 lysis buffer [50 mM Tris (pH 7.5), 0 mM MgCl2, 1 % Triton X-100, 1 mM dithiothreitol, 1 mM EDTA, 1 mM EGTA, with freshly added 1 mM β - glycerophosphate and protease inhibitors (phenylmethylsulfonyl fluoride and aprotinin)-RSB- and immunoprecipitated overnight using 1.5 μg rabbit anti-RIPK2 antibody.
Cells were then
lysed and probed for transcription of Nrf2, Keap1, or Bach1 using the QuantiGene II RNA plex (Affymetrix).
Cell supernatants were discarded and cell pellets were
lysed of RBCs and then used for staining with antibodies for flow cytometry analysis.
For Western blot analysis, 106 BaF3 cells were starved for 4 h and
lysed in 250 μL of Laemmli buffer (BioRad).
Cells were
lysed with 150 μl / well of RIPA buffer.
Cells were then
lysed and HMOX1 mRNA was detected using the Quantigene II method as described in Materials and Methods.
Cells were
lysed with TN1 lysis buffer (50 mM Tris, 125 mM NaCl, 1 % Triton X-100, 10 mM EDTA, 10 mM sodium fluoride and 10 mM sodium pyrophosphate) supplemented with protease inhibitor cocktail (1.2 mM AEBSF, 0.46 μM aprotinin, 14 μM bestatin, 12.3 μM E-64, 112 μM leupeptin, 1.16 μM pepstatin)(Amresco; Solon, OH)-RRB- and 1 mM sodium orthovanadate (Enzo Life Sciences; Farmingdale, NY).
NHLF cells transfected with siRNA molecules were
lysed in High Salt ELB lysis buffer [1 M Tris pH 8.0, 1 % NP - 40, 250 mM NaCl, 5 mM EDTA] supplemented with protease and phosphatase inhibitors (1x G - Biosciences Protease Arrest, 200 µM Na3VO4, and 1 mM PMSF).
Cells were
lysed in radioimmunoprecipitation assay buffer with protease and phosphatase inhibitors.
Cells were
lysed in RIPA lysis buffer.
Cells were washed once with PBS and
lysed in 0.02 mM NaOH for 16 hr at 4 °C, and then stored at — 20 °C until analysis.
Cells were
lysed on ice using lysis buffer (10 mM Tris - HCl pH 7.4, 150 mM NaCl, 1 % Triton X-100, protease inhibitor mix G (Serva) and 2 mM Na3VO4).
Spleen cells (3 × 107) from either experimental or control effector lymphocytes were dispensed into 75 - cm2 tissue culture flasks along with 20 ml of boosting medium and 6 × 105 γ - irradiated (3,000 rad) BALB / c spleen cells (erythrocytes
lysed).
At 48 hours, cells were washed twice with PBS and
lysed in 1 × passive lysis buffer (Dual - Luciferase Reporter Assay, Promega).
Additionally, most often detection is carried out on
lysed cell samples, so a significant amount of information is lost.
Cells were
lysed and membrane and cytosolic fractions separated and EGFR abundance determined by Western blot.
EGFR / WT and KO MEFs were treated with capsaicin (0, 10, or 50 μmol / L) for 30 min followed by TPA (20 ng / mL) for 4 h. Cells were
lysed and COX - 2 expression analyzed by Western blot.
EGFR / WT and KO cells were cultured overnight and starved for 24 h. Cells were treated with capsaicin (50 μmol / L) for 30 min and then TPA (20 ng / mL) for 2 h. Cells were
lysed and cox - 2 mRNA levels were analyzed by RT - PCR.
In a new report, the BBC reporter
Lyse Doucet and camera operator Philip Goodwin described the atmosphere at the luxury hotel as «very serious.»
One device segregates the cells into individual gel droplets, allowing researchers to
lyse the cells and amplify their DNA while keeping them separated.
Lyse, my little friends,
lyse!»
Lysing the red blood cells diminishes the tendency of blood to clog the system, a common problem that slows processing time in similar CTC filtering devices.
We have designed a microfluidic device in which we can manipulate,
lyse, label, separate, and quantify the protein contents of a single cell using single - molecule fluorescence counting.
The distinction between apoptosis and cell lysis is important in immunology -
lysing a virus - infected cell would only release the virions, whereas apoptosis leads to destruction of the virus inside.
The metallopolymer by itself even demonstrated antimicrobial properties,
lysing bacterial cells while leaving human red blood cells unaffected.
An examination into the host's immune response to symbiont presence suggests that firebugs actively harvest their bacterial partners by
lysing, or bursting open, the bacterial cells.
In addition, it was increasingly recognized that oncolytic viruses not only were able to directly
lyse cancer cells, but they also «freed «tumor specific neoantigens, indirectly acting as a cancer vaccine.
Several groups are now trying to take advantage of the tumor -
lysing capacity of viruses to make better vaccines.
The usual way is to
lyse the cell, or break it open so that its contents spill out.
NK cells are able to
lyse virally infected cells and secrete large amounts of proinflammatory cytokines / chemokines without prior antigen sensitization.
PC3 and PC3 DAXX K / D cells were processed using Power SYBR Green Cells - to - Ct Kit (Ambion, Cat # 4402953) to
lyse cells, generate cDNA, and perform Q - PCR per the manufacturer's instructions.