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Pbs based cell dissociation buffer

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The PBS-based cell dissociation buffer is a laboratory reagent designed to facilitate the detachment of adherent cells from cell culture surfaces. It contains a balanced salt solution and chelating agents that help break down the cell-to-surface adhesions, enabling the gentle and efficient release of cells without compromising their viability or functionality.

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Lab products found in correlation

Facscalibur flow cytometer, by BD (2 mentions) Calcein am, by BioLegend (1 mentions) Lsrfortessa, by BD (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Annexin 5 pe, by BD (1 mentions) Low binding plates, by Corning (1 mentions) Facscalibur system, by BD (1 mentions) Cellquest pro software, by BD (1 mentions) Goat anti human igg fitc, by Jackson ImmunoResearch (1 mentions) Collagenase type 1, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

Close spelling variants of this product

Cell Dissociation Buffer (322 citations) PBS buffer (120 citations) Dissociation buffer (18 citations) Cell Dissociation Buffer Enzyme Free PBS-Based (13 citations) Cell Dissociation Buffer—PBS (4 citations)

9 protocols using pbs based cell dissociation buffer

1

Photothermal Cytotoxicity of LPNs

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The Hs578T and T47D cells were seeded on a 24-well plate at a density of 2 × 105 cells/well. After a 48 h incubation, cells were treated with 300 μg/mL of IG-LPNs or C-LPNs for 1 h at 37 °C. Thereafter, cells were detached with enzyme-free, PBS-based cell dissociation buffer (Thermo Fisher Scientific, Inc.) and centrifuged at 3000 rpm for 5 min (Merck Millipore). The cell pellets were irradiated for 5 min with an NIR laser (808 nm) at 1.5 W power using a diode laser beam (BWT Beijing Ltd.). Temperature was measured using an infrared camera (FLIR E60). The irradiated cells were seeded on a 96-well plate and incubated for 24 h at 37 °C. The viability of cells was measured using a water soluble tetrazolium salt (WST) assay and visualized by staining live cells and dead cells using calcein-AM (Biolegend Inc.) and propidium iodide (Biomax, Gongneung-dong, Seoul, Republic of Korea), respectively.
Yang H., Le Q.V., Shim G., Oh Y.K, & Shin Y.K. (2020). Molecular engineering of antibodies for site-specific conjugation to lipid polydopamine hybrid nanoparticles. Acta Pharmaceutica Sinica. B, 10(11), 2212-2226.
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2

Microglia Uptake and Degradation of Amyloid-Beta

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Primary microglia or MDMs were serum starved overnight before the addition of Ab, Ab-LDL, or Ab-CLU. Following a period of time for Ab uptake, cells were dislodged using enzyme-free PBS-based cell dissociation buffer (Thermo Fisher Scientific). Cells were washed and resuspended in flow cytometry buffer (13 PBS, 0.5% BSA, and 0.05% sodium azide) and kept on ice. Sytox blue staining was used to identify live cells. Flow cytometry was done using LSRFortessa (BD Biosciences) and the mean FITC/DiI fluorescence intensity was analyzed.
For Ab degradation experiments, microglia cultures were incubated with free Ab aggregates (0.1 mM) or Ab-LDL complexes (0.01 mM Ab) for 2 hr, after which they were washed with media and placed back into fresh media. Cells were lysed at 0, 1, 3, 5, or 7 hr, and the amount of remaining Ab in the cell lysate was measured by Meso Scale Diagnostics immunoassays.
Yeh F.L., Wang Y., Tom I., Gonzalez L.C, & Sheng M. (2016). TREM2 Binds to Apolipoproteins, Including APOE and CLU/APOJ, and Thereby Facilitates Uptake of Amyloid-Beta by Microglia. Neuron, 91(2).
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3

Co-culture Assay for mSiglec1 Expression

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HEK293T cells were grown in DMEM with 10% FBS, and 70–80% confluent cells were used for transfection with mammalian expression plasmids pd18 (empty plasmid; mock) or pd18-mSiglec1 using Lipofectamine 3000 (Invitrogen). Medium was changed to CDM293 (Invitrogen) supplemented with L-glutamine and cells were further cultured for 3 days to ensure maximum mSiglec1 expression on the cell surface. On day 3, single-cell preparations of B16-GFP cells were prepared by dissociating cells with enzyme-free PBS-based cell dissociation buffer (Gibco) and passing cells through 40 μm nylon filters (Falcon). Both HEK293T cells and cancer cells were counted and mixed at a 4:1 ratio in DMEM supplemented with 5% FBS. Cells were cultured in low binding plates (Corning) for 18 hr at 37°C in 5% CO2. At the end of the experiments, cells either underwent cell sorting to assess gene expression or were fixed in Cytofix buffer for Ki67 or Phosflow staining. For apoptosis quantification PE Annexin V (BD biosciences) was used according to manufacturer’s instructions.
Singh R, & Choi B.K. (2019). Siglec1-expressing subcapsular sinus macrophages provide soil for melanoma lymph node metastasis. eLife, 8, e48916.
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4

h4G3 Binding to CLDN in Cells

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To analyze h4G3 binding to CLDN, cells were detached with enzyme-free, PBS-based cell dissociation buffer (Gibco). A total of 2.5 × 105 cells were incubated with human IgG (10 μg/mL) (Jackson Immunoresearch Laboratories, West Grove, PA, USA) or h4G3 (10 μg/mL) in PBS containing 1% FBS for 1 h on ice. The cells were then washed three times with PBS containing 1% FBS and incubated with goat anti-human IgG-FITC (Jackson Immunoresearch Laboratories) (1:100 dilution) for 1 h on ice. Stained cells were washed three times and analyzed using a BD FACSCalibur system equipped with the Cell Quest Pro software (BD Biosciences).
Yang H., Park H., Lee Y.J., Choi J.Y., Kim T., Rajasekaran N., Lee S., Song K., Hong S., Choi J.S., Shim H., Kim Y.D., Hwang S., Choi Y.L, & Shin Y.K. (2019). Development of Human Monoclonal Antibody for Claudin-3 Overexpressing Carcinoma Targeting. Biomolecules, 10(1), 51.
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5

Flow Cytometry Analysis of PD-L1 Expression

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Adherent cells were detached using enzyme-free, PBS-based cell dissociation buffer (Gibco). The harvested cells were washed twice with flow cytometry buffer (1× PBS with 2 mM EDTA and 0.5% FBS) and stained with anti-human PD-L1 antibody conjugated with phycoerythrin (PE) (catalog #557924, BD) according to the manufacturer’s protocol.
For harvesting cells from the 4T1-Luc tumor, the tumor was dissected out, cut into small pieces, minced finely with scissors and scalpel, and then incubated in Hanks’ balanced salt solution (HBSS) containing 1 mg/mL of collagenase type I (Gibco) for 30 min at 37 °C. All of the digested tumor mass was passed through a 70 µM cell strainer to obtain a suspension of single cells. This suspension was then washed twice with flow cytometry buffer and then stained with anti-mouse PD-L1 antibody conjugated with phycoerythrin (PE) (catalog #558091, clone MIH5, BD) according to the manufacturer’s protocol.
Samples were aquired on a FACSCalibur flow cytometer (Becton Dickinson). At least 20,000 events were recorded and analyzed using FlowJo software (Tree Star).
Lesniak W.G., Chatterjee S., Gabrielson M., Lisok A., Wharram B., Pomper M.G, & Nimmagadda S. (2016). PD-L1 Detection in Tumors Using [64Cu]Atezolizumab with PET. Bioconjugate chemistry, 27(9), 2103-2110.
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6

CXCR4 and PDGFRα Expression Analysis

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For CXCR4 and PDGFRα analysis, cells were dissociated with PBS‐based cell dissociation buffer (Gibco), resuspended in PBS/1% rat serum and incubated with APC‐conjugated CXCR4 antibody or APC‐conjugated PDGFRα antibody (details of antibodies used are listed in Table EV1) and analysed on a CyAn (Beckman Coulter). DAPI counterstaining was used to exclude dead cells.
Morrison G., Scognamiglio R., Trumpp A, & Smith A. (2015). Convergence of cMyc and β‐catenin on Tcf7l1 enables endoderm specification. The EMBO Journal, 35(3), 356-368.
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7

Isolation and Characterization of MSCs

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MSCs and patient-derived MSCs were harvested with PBS-based cell dissociation buffer (Gibco; 1 × 105 cells per tube), washed, suspended in PBS with 5% BSA, and stained with marker-specific fluorochrome-conjugated antibodies for 30–60 min in the dark. Nonspecific background signals were measured by incubating separate tubes with appropriate isotype-matched non-specific antibodies. Cells were then resuspended in PBS + 5% BSA and filtered through a 40μm cell strainer (Flowmi) and sorted by a LSRII flow cytometer (BD Biosciences). Cell debris and clumps were electronically gated from analysis based on their forward and side light scatter parameters.
Gordon J.A., Evans M.F., Ghule P.N., Lee K., Vacek P., Sprague B.L., Weaver D.L., Stein G.S, & Stein J.L. (2023). Identification of molecularly unique tumor-associated mesenchymal stromal cells in breast cancer patients. PLOS ONE, 18(3), e0282473.
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8

PD-L1 Flow Cytometry Assay

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Cells in suspension were harvested by centrifugation with adherent cells detached using enzyme-free, PBS-based cell dissociation buffer (Gibco). The harvested cells were washed twice with flow cytometry buffer (1xPBS with 2 mM EDTA and 0.5% FBS). Cells were stained with anti-human PD-L1 antibody conjugated with phycoerythrin (PE) (catalog #557924, Becton Dickinson) according to the manufacturer's protocol and were analyzed on a FACSCalibur flow cytometer (Becton Dickinson). At least 20,000 events were recorded and analyzed using FlowJo software (Tree Star).
Chatterjee S., Lesniak W.G., Gabrielson M., Lisok A., Wharram B., Sysa-Shah P., Azad B.B., Pomper M.G, & Nimmagadda S. (2016). A humanized antibody for imaging immune checkpoint ligand PD-L1 expression in tumors. Oncotarget, 7(9), 10215-10227.
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9

CXCR4 Expression in Gemcitabine-Treated Cells

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CD18/HPAF.luc cells were seeded in 6-well plates in DMEM with 10% FBS. The next day, fresh DMEM with or without 10 μM gemcitabine was added. After 24 h of incubation, cells were washed with PBS twice, detached using enzyme-free PBS-based cell dissociation buffer (Gibco by Life Technologies, Grand Island, NY), and resuspended in PBS. After centrifugation at 800 rpm for 5 min, the supernatant was removed and the cells were resuspended in FACS buffer (1% BSA, 0.1% NaN3 in PBS) to obtain final cell concentration of 5 × 106/mL. The cell suspension (100 μL) was mixed with 20 μL of APC mouse anti-human CXCR4 antibody or the corresponding isotype control. After incubation at room temperature for 30 min, the cells were washed three times with FACS buffer to remove free antibody and resuspended in 500 μL of FACS buffer for flow cytometry. The data were processed and analyzed using FlowJo software (Tree Star Inc., Ashland, OR) and reported as percentage of CXCR4-positive cells.
Wang Y., Kumar S., Rachagani S., Sajja B.R., Xie Y., Hang Y., Jain M., Li J., Boska M.D., Batra S.K, & Oupický D. (2016). Polyplex-mediated inhibition of chemokine receptor CXCR4 and chromatin-remodeling enzyme NCOA3 impedes pancreatic cancer progression and metastasis. Biomaterials, 101, 108-120.
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