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Lyse buffer

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Lyse buffer is a solution used in laboratory procedures for the lysis or disruption of cells, tissues, or other biological samples. It is designed to efficiently release the contents of cells, including proteins, nucleic acids, and other cellular components, without damaging the target molecules. The core function of the lyse buffer is to facilitate the extraction and isolation of these cellular components for further analysis or experimentation.

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Lab products found in correlation

Moflo xdp cell sorter, by Beckman Coulter (1 mentions) Macs inside stain kit, by Miltenyi Biotec (1 mentions) Lsrii cytometer, by BD (1 mentions) Siglec f, by BD (1 mentions) Gammacell 40 irradiator, by Nordion (1 mentions)

Close spelling variants of this product

Lyse/Fix and Perm III buffers FACS lyses buffer BD Pharm Lyse lysing buffer BD Phosflow Lyse/Fix Buffer Lyse/fix Buffer 5X BD Pharm Lyse lysis buffer BD Phosflow Lyse/Fix Buffer 5 × Lyse/Fix Buffer 5× Lyse/Fix Buffer Phosflow Lyse/Fix Buffer 5X

4 protocols using lyse buffer

1

Complement C5a Quantification from Conditioned Media

Cited 2 times
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BMMNCs from WT or Casp1-KO or NLRP3 KO mice injected with the mixture of interleukins or interleukins and DAMPs in the presence or absence of AMD3100 or G-CSF were isolated as described above. The residual RBCs were lysed using Lyse buffer (BD Biosciences, San Jose, CA, USA). Then, the samples were incubated in 0.5% BSA containing RPMI for 24 h at 37 °C in a 5% CO2 incubator. The culture supernatant was collected. A 100 µl of the conditioned medium samples were used in this experiment [24 (link), 28 (link)]. The experiments were performed in triplicates using complement C5a mouse ELISA kit (Abcam, Cambridge, MA, USA) as described in manufacturer’s protocol.
Thapa A., Adamiak M., Bujko K., Ratajczak J., Abdel-Latif A.K., Kucia M, & Ratajczak M.Z. (2021). Danger-associated molecular pattern molecules take unexpectedly a central stage in Nlrp3 inflammasome–caspase-1-mediated trafficking of hematopoietic stem/progenitor cells. Leukemia, 35(9), 2658-2671.
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2

Purification of SKL cells from mouse bone marrow

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SKL cells were purified from the bone marrow (BM)-derived total nucleated cells (TNCs) of C57BL/6 wild-type and Nox2-knock out mice. In brief, BM was flushed from the femurs, and the population of TNCs was retrieved after lysis of red blood cells (RBCs) using Lyse buffer (BD Pharmingen, San Jose, CA, USA). Afterward, the harvested cells underwent staining with the following antibodies: lineage marker-specific antibodies: PE–anti-TCRγδ, clone GL3; PE–anti-CD11b, clone M1/70; PE–anti-TCR β-chain, clone H57–597; PE–anti-CD45R/B220, clone RA3-6B2; PE–anti-TER-119/erythroid cells, clone TER-119; PE–anti-Ly-6G and -Ly-6C (Gr1), clone RB6-8C5; PE–Cy5–anti-Ly-6A/E (Sca-1), clone D7; fluorescein isothiocyanate (FITC)–anti-CD117 (c-Kit), clone 2B8 in RPMI-1640 medium supplemented with 2% fetal bovine serum (FBS). All these antibodies were purchased from BD Biosciences. After 30 min of staining, the cells were washed, resuspended, and sorted using a Moflo XDP cell sorter (Beckman Coulter, Indianapolis, IN, USA) as populations of SKL (Sca-1+c-Kit+ Lin−) cells [18 (link)].
Abdelbaset-Ismail A., Ciechanowicz A.K., Bujko K., Ratajczak J., Kucia M, & Ratajczak M.Z. (2022). The Nox2-ROS-Nlrp3 Inflammasome Signaling Stimulates in the Hematopoietic Stem/Progenitor Cells Lipogenesis to Facilitate Membrane Lipid Raft Formation. Stem Cell Reviews and Reports, 19(1), 92-103.
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3

Flow Cytometry Analysis of Stromal Vascular Fraction

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Following isolation, the fresh SVF from young and aged donors was incubated in rat Fc Block Mix (BD Biosciences 550271) at 4°C for 20 min protected from light. Cells were then stained for cell surface markers with the appropriate monoclonal antibodies at 4°C for 30 min also protected from light. Supplemental Table 1 contains the antibodies included in each staining panel used in this study along with dilutions. During the incubation, anti-mouse IgG kappa compensation beads and negative control beads (BD Biosciences 552843) were added to each compensation tube; antibodies (positive compensation) or fluorophore-conjugated IgG (negative compensation) were added to the appropriate tubes. Following antibody incubation, red blood cells were lysed with diluted Lyse buffer (BD Bioscience 5558999) and tubes were vortexed, incubated for 3 min at 37°C, spun at 350 g for 5 min, then cells washed twice with wash buffer. For the tubes requiring CD68 staining, cells were fixed and stained with MACS Inside Stain Kit (Miltenyi Biotec, 130–090-477) according to manufacturer’s instructions. Data was collected on an LSR II cytometer (BD Biosciences) and analyzed with FlowJo 7.6 software (Tree Star, Ashland, OR, USA).
Rowe G., Heng D.S., Beare J.E., Hodges N.A., Tracy E.P., Murfee W.L, & LeBlanc A.J. (2022). Stromal Vascular Fraction Reverses the Age-Related Impairment in Revascularization Following Injury. Journal of vascular research, 59(6), 343-357.
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4

Hematopoietic Stem Cell Transplantation

Cited 1 time
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Bone marrow was aseptically harvested from tibias, femurs and humeri from 9-week-old mice, erythrocytes lysed (BD Pharm Lyse buffer) and cells incubated with Fc-block followed by incubation with fluorochrome-conjugated antibodies against B220, CD4, CD8, CD11b, Ly6G, NK1.1, Siglec F, Ter119, c-Kit and Sca-1 (BD Biosciences, eBioscience, Biolegend), as previously described [38 (link)]. Cell suspensions were sorted by fluorescence-activated cell sorting (FACS) to obtain the Lin-Sca-1+c-kit + (LSK) population. Two-month-old B6.CD45.1 mice received a single 1000-cGy γ-irradiation dose using a Cs-137-based Gammacell-40 irradiator (Nordion). After 12 hours, 5 × 104 LSK cells were intravenously injected from: Casp8flox/flox, CreCD11cCasp8flox/flox, RIPK3–/–CreCD11cCasp8flox/flox, a mixture of Casp8flox/flox plus B6.CD45.1/2, CreCD11cCasp8flox/flox plus B6.CD45.1/2 or RIPK3–/–CreCD11cCasp8flox/flox plus B6.CD45.1/2 (1:1 ratio). Chimeric mice were maintained on Trimetoprim/Sulfamethoxazole (40 mg/5 mg, respectively; Hi-Tech Pharmacal) diluted in autoclaved water (2 mL antibiotics/500 mL water) and phenotyped 2 months post transfer.
Dominguez S., Montgomery A.B., Haines GK I.I.I., Bloomfield C.L, & Cuda C.M. (2017). The caspase-8/RIPK3 signaling axis in antigen presenting cells controls the inflammatory arthritic response. Arthritis Research & Therapy, 19, 224.
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