Log In Sign up
The largest database of trusted experimental protocols

Carboxyfluorescein succinimidyl ester (cfse)

Manufactured by R&D Systems
Visit Supplier
Sourced in United States

CFSE is a cell labeling dye that can be used to track cell division and proliferation. It binds to cellular proteins, and its fluorescence intensity is reduced by half with each cell division, allowing for the monitoring of cell division history.

Automatically generated - may contain errors

Lab products found in correlation

Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (13 mentions) Interleukin 2 (il 2), by R&D Systems (3 mentions) Htgf β, by R&D Systems (2 mentions) Cd4 t cell macs separation kit, by Miltenyi Biotec (2 mentions) Anti mouse antibodies, by BD (2 mentions) Anti cd3e, by Thermo Fisher Scientific (2 mentions) Easysep mouse biotin positive selection kit, by STEMCELL (2 mentions) Gr 1 pe, by Thermo Fisher Scientific (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Lymphocyte separation fluid, by Dakewe (1 mentions) Anti cd3, by R&D Systems (1 mentions) Anti cd28, by R&D Systems (1 mentions) Il 23 mouse fc, by Adipogen (1 mentions) Cd4 microbeads, by Miltenyi Biotec (1 mentions) Brefeldin a, by Merck Group (1 mentions) Percp conjugated anti cd3, by BD (1 mentions) Apc h7 conjugated cd8, by BD (1 mentions) Brilliant violet 650 conjugated cd4, by BioLegend (1 mentions) Alexa 700 conjugated cd14 and cd19, by BioLegend (1 mentions) Apc conjugated anti γδ, by BioLegend (1 mentions)

15 protocols using carboxyfluorescein succinimidyl ester (cfse)

1

Purifying Naive CD4+ T Cells and Dendritic Cells for In Vitro Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
CD11chighMHCIIhigh DCs were purified from lymph nodes of C57BL/6 and Batf3−/− mice as described above. For the isolation of OVA-specific naive T cells, spleens were obtained from C57BL/6 Rag1−/− OT-II Tg mice. Naive CD4+ T cells were enriched using CD4+ T cell MACS separation kit (Miltenyi Biotech) and subsequently FACSsorted based on the expression of CD4, CD45RB and CD25 to obtain >99% pure population. The purified naive OVA-specific CD4+ T cells were labeled with CFSE (Invitrogen) at a final concentration of 2 μM. For the in vitro stimulation assays, dendritic cells were cultured with 2 mg/ml OVA protein for 1 hours at 37 °C. Subsequently, cells were washed extensively with sterile PBS after which the CFSE-labeled T cells were added at a 1:10 ratio in the presence or absence of 2 ng/ml hTGFβ (R&D Systems Inc). At 72 hours, cells were analyzed for division and Foxp3 expression.
Veenbergen S., van Berkel L.A., du Pré M.F., He J., Karrich J.J., Costes L.M., Luk F., Simons-Oosterhuis Y., Raatgeep R.H., Cerovic V., Cupedo T., McI Mowat A., Kelsall B.L, & Samsom J.N. (2015). Colonic tolerance develops in the iliac lymph nodes and can be established independent of CD103+ dendritic cells. Mucosal immunology, 9(4), 894-906.
+ Open protocol
+ Expand
2

Modulation of T Cell Proliferation by Neutrophils and Tumor Secretome

Check if the same lab product or an alternative is used in the 5 most similar protocols
Spleens were removed from 8-week female C57BL/6 wild-type mice of and placed in a 70 μm cell sieve and gently grind the spleen until no obvious tissue mass was seen. The n 4–5 ml of lymphocyte separation fluid (Dakewe, China) was added to re-suspend the tissues. The cell suspension was placed in a 15 ml centrifuge tube for gradient centrifugation at 800 g for 30 min. Lymphocytes were purified from the liquid and stained with CFSE (Invitrogen). The CFSE-labelled lymphocytes were placed in 24-well plates supplemented complete RPMI medium with 1 μg/ml anti-CD3 (R&D Systems) and 5 μg/ml anti-CD28 (R&D Systems) antibodies. Isolated neutrophils (1:1) or tumor supernatant (50%, v/v) were added to the 24-well plate. After incubation for 72 h, T cells were collected and stained with anti-CD3, anti-CD4, and anti-CD8 antibodies for flow cytometric analysis.
Cheng Y., Mo F., Li Q., Han X., Shi H., Chen S., Wei Y, & Wei X. (2021). Targeting CXCR2 inhibits the progression of lung cancer and promotes therapeutic effect of cisplatin. Molecular Cancer, 20, 62.
+ Open protocol
+ Expand
3

Adoptive Transfer of Splenic Thy1.1+ DO11.10 CD4+ T Cells to Investigate Irf5 Role

Check if the same lab product or an alternative is used in the 5 most similar protocols
Splenic Thy1.1+ DO11.10 CD4+ T cells were isolated
using CD4+ MicroBeads (Miltenyi Biotec), stained with 1.5–2.5
μM CFSE (Molecular Probes), and adoptively transferred by i.v. injection
(0.5 × 106) into Thy1.2+Irf5+/+, Irf5+/−,
or Irf5−/− BALB/c mice. Twenty-four
hours later, mice were immunized with s.c. 25 μg chicken OVA (MP
Biomedicals) and 25 μg LPS (MilliporeSigma). Three days after
immunization, peripheral lymph nodes (PLN) were harvested and stained with Abs
to CD4, Thy1.1, and DO11.10. In other experiments, serum was harvested 2 wk
after immunization for measurement of chicken OVA-specific isotype Abs.
Ninety-six–well plates were coated with OVA (50 μg/ml) in PBS and
plates ultimately incubated with IgG2a-, IgG2b-, or IgG1-conjugated with
alkaline phosphatase (SouthernBiotech). For ex vivo detection of intracellular
cytokines, cells were first treated with 2 μg/ml OVA peptide
(323–339) (LifeTein) for 12 h and 10 μg/ml brefeldin A
(MilliporeSigma) was added in the last 10 h. In some experiments, CFSE-labeled
splenic Thy1.1+ DO11.10 CD4+ T cells (0.5 ×
106) were first coated with 20 μg anti-mouse CCR7
neutralizing Ab (R&D Systems) for 30 min. In other experiments, 100 ng/mouse
IL-12 (mouse)/Fc and 500 ng/mouse IL-23 (mouse)/Fc (AdipoGen Life Sciences) or
Fc isotype controls (Bio X Cell) were injected i.v. into mice prior to
immunization.
Yan J., Hedl M, & Abraham C. (2020). Myeloid Cell–Intrinsic IRF5 Promotes T Cell Responses through Multiple Distinct Checkpoints In Vivo, and IRF5 Immune-Mediated Disease Risk Variants Modulate These Myeloid Cell Functions. Journal of immunology (Baltimore, Md. : 1950), 205(4), 1024-1038.
+ Open protocol
+ Expand
4

CFSE-based T cell proliferation assay with PBMC

Check if the same lab product or an alternative is used in the 5 most similar protocols
Thawed PBMC were rested for one hour, washed in 10% Human AB media (Gemini), and 3–6×106 PBMC were labeled with 1 ml of 1.25 µM 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE; Molecular Probes) for seven minutes. CFSE-labeled PBMC were incubated in 96-well, deep-well culture plates (Nunc, Roskilde, Denmark) at a density of 106 PBMC per well at a final volume of 1 ml for 7 days. In a subset of patients, CFSE-labeled PBMC were incubated with antigen in the presence of IL-10 receptor alPHA chain (IL-10Rα) blocking antibody (clone 37607; R&D Systems) or IgG1 isotype control antibody at 10 µg/mL. Antigens tested included media, phytohemagglutinin (PHA; 5 µg/mL; Sigma-Aldrich), uRBC, or PfSE at an E:T ratio of 1∶3 schizont equivalents. At day 7 cells were treated with 100 units DNase I (Invitrogen) in culture medium at 37°C for 10 min, washed, and stained with surface antibodies (PerCP–conjugated anti-CD3, APC-H7-conjugated CD8 (BD PHArmingen), Brilliant violet 650-conjugated CD4, Alexa 700-conjugated CD14 and CD19, and APC-conjugated anti-γδ (Biolegend)) before acquisition.
Jagannathan P., Eccles-James I., Bowen K., Nankya F., Auma A., Wamala S., Ebusu C., Muhindo M.K., Arinaitwe E., Briggs J., Greenhouse B., Tappero J.W., Kamya M.R., Dorsey G, & Feeney M.E. (2014). IFNγ/IL-10 Co-producing Cells Dominate the CD4 Response to Malaria in Highly Exposed Children. PLoS Pathogens, 10(1), e1003864.
+ Open protocol
+ Expand
5

Modulating CD8+ T Cell Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols
A 24-well plate was coated with 1 μg/well of the anti-CD3e antibody. CD8+ T cells were purified from the spleen of a Balb/c mouse using T-cell isolation columns (R&D systems, Minneapolis, MN, USA) according to the manufacturer's instructions and labeled with carboxyfluorescein succinimidyl ester (CFSE). In order to assess the impact of MDSC on CD8+ T cell proliferation, CD8+ T cells were co-cultured with MDSC with or without E2, ICI 182,780, and JSI-124 for 48 hours. T-cell proliferation was evaluated by flow cytometry. CFSE was purchased from Tonbo Biosciences (San Diego, CA, USA).
Kozasa K., Mabuchi S., Matsumoto Y., Kuroda H., Yokoi E., Komura N., Kawano M., Takahashi R., Sasano T., Shimura K., Kodama M., Hashimoto K., Sawada K., Nagasaka K, & Kimura T. (2019). Estrogen stimulates female cancer progression by inducing myeloid-derived suppressive cells: investigations on pregnant and non-pregnant experimental models. Oncotarget, 10(20), 1887-1902.
+ Open protocol
+ Expand
6

T Cell Proliferation Assay with Myeloid Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
CD3+ T cells of naïve BALB/c or C57BL/6 mice (6 – 8 weeks old) were enriched from spleen by negative selection using biotinylated anti-mouse antibodies against B220, CD11b, Gr1 (BD Pharmingen, cat # 559971) and CD11c (BD Pharmingen, cat # 553800) followed by magnetic separation using EasySep Mouse Biotin Positive Selection Kit (STEMCELL Technologies). Bone marrow neutrophils and monocytes from either naïve or relevant tumor-bearing animals were harvested as previously described in “In vitro trans-well migration assay”. Magnetically sorted CD3+ T cells were labeled with CFSE (5 μM, Molecular Probes) as per manufacturer’s instruction. T cells were cultured alone or admixed with neutrophils or monocytes (at 1:3 ratio) in 96 well plate. T cell activation was with anti-CD3e (eBioscience, cat # 16–0031-85) through coating of wells at 5 μg/mL, overnight, 4°C and IL-2 (R&D, cat # 202-IL-010/CF) at 5 ng/mL. After 4 days of co-culture, cells were collected and analyzed for CFSE intensity by flow cytometry. Collected cells were also stained for Gr1-PE (ebioscience, cat # 12–5931-82) to be able to exclude non-T cells from the analysis. Proliferation index (%) was calculated as follows: (percentage of proliferated, co-cultured CD3+ T cells)/(percentage of proliferated CD3+ T cells cultured alone) X 100.
Kim I.S., Gao Y., Welte T., Wang H., Liu J., Janghorban M., Sheng K., Niu Y., Goldstein A., Zhao N., Bado I., Lo H.C., Toneff M.J., Nguyen T., Bu W., Jiang W., Arnold J., Gu F., He J., Jebakumar D., Walker K., Li Y., Mo Q., Westbrook T.F., Zong C., Rao A., Sreekumar A., Rosen J.M, & Zhang X.H. (2019). Immuno-subtyping of breast cancer reveals distinct myeloid cell profiles and immunotherapy resistance mechanisms. Nature cell biology, 21(9), 1113-1126.
+ Open protocol
+ Expand
7

Dendritic Cell Activation and T Cell Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
DC (2 × 106 cells/ml) were cultured in complete RPMI 1640 supplemented with 10% FCS alone or in the presence of R837 (0.5 μg/ml), LPS (5 μg/ml), CpGA-ODN1585 (5 μg/ml), or polyinosinic:polycytidylic acid [poly(I:C); 5 μg/ml] (all from InvivoGen, San Diego, CA). Cytokine secretion was determined at 20 h of culture by ELISA. For co-culture experiments, MACS purified (Miltenyi Biotech, San Diego, CA, USA) CD4+ T cells were labeled with CFSE (1 μM; R&D Systems; Minneapolis, MN, USA) for 5 min at RT, followed by extensive washing in complete RPMI 1640, prior to culture. DC were cultured at a ratio of 1:20 with CD4+ T cells (1.25 × 105/ml) in the presence of anti-CD3 (1 μg/ml). Cells were collected on day 3 of culture and stained with CD4 antibody for detection of T cells and analyzed by Flow cytometry using FACSDiva Software on an LSRII (BD Biosciences; San Jose, CA, USA).
Tu H., Burke T.M., Oderup C., Huang K., Wong K., Lewén S., LaJevic M, & Zabel B.A. (2014). Robust Expansion of Dendritic Cells in vivo by Hydrodynamic FLT3L-FC Gene Transfer. Journal of immunological methods, 0, 69-73.
+ Open protocol
+ Expand
8

B Cell Isotype Switching and KSHV Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse B cells were purified from freshly isolated splenocytes using anti-CD43 magnetic beads MACS CD43 depletion (Miltenyi Biotech) according to manufacturer’s protocol. Cells were stained with CFSE (Invitrogen) or eFlour670 (Thermo Fisher Scientific) and 5 × 105 cells/mL were activated with 5 μg/mL LPS (Sigma) and 20 ng/mL murine IL-4 (Peprotech) (IgG1 switching), or 1 ng/mL TGF-β1 (IgG2b switching). Isotype switching was analyzed by FACS after staining cells with anti–IgG1 or IgG2b-biotin (BD Pharmingen), followed by PE-conjugated anti-biotin antibody. Splenocytes were cultured with or without purified rKSHV.219 (MOI = 1–5) with 5 μg/mL protamine sulfate and activated with the aforementioned cytokines. For CH12F3-2 experiments, cells were also preincubated with CFSE or eFluor 670 and 25 × 104 cells/mL were activated with 1 ng/mL TGF-β1 (R&D Systems), 10 ng/mL recombinant murine IL-4 (PeproTech), and 1 μg/mL purified anti–mouse CD40 (BD Biosciences). Cells were stained with anti-IgA (SouthernBiotech) and analyzed by FACS with Accuri C6 Flow Cytometer (BD Biosciences).
Rosario S.A., Santiago G.E., Mesri E.A, & Verdun R.E. (2018). Kaposi’s Sarcoma-Associated Herpesvirus-Encoded Viral IL-6 (vIL-6) Enhances Immunoglobulin Class-Switch Recombination. Frontiers in Microbiology, 9, 3119.
+ Open protocol
+ Expand
9

Suppression of PBMC Proliferation by MDSC

Check if the same lab product or an alternative is used in the 5 most similar protocols
MDSC were generated from myeloid cells of the PBMC fraction as described above and isolated from cell cultures by magnetic bead cell sorting for CD33 (Miltenyi Biotec). Responder-PBMC were obtained from healthy volunteers' heparinized blood and stained with CFSE (Life Technologies) according to the manufacturer's protocol. CFSE-labeled PBMC were stimulated with 100 U/ml IL-2 (R&D Systems) and 1 μg/ml OKT3 (Janssen-Cilag). Both MDSC and CFSE-labeled PBMC were added to RPMI 1640 medium supplemented with 10% human serum, 2 mM L-glutamine, 100 IU/ml penicillin and 100 mg/ml streptomycin. In a 96-well round bottom plate (Greiner Bio-One), either 10,000/30,000 MDSC or, as a control supplemented medium only, were added to 60,000 PBMC per well. Cells were incubated in a humidified atmosphere at 37°C and 5% CO2. On day 4 cells were harvested and stained with anti-CD8a-APC, anti-CD4-PE antibodies (BioLegend), and propidium iodide (BD). PI positive cells were excluded in flow cytometry. CFSE signals of CD4+ and CD8+ PBMC were analyzed.
Stoll H., Ost M., Singh A., Mehling R., Neri D., Schäfer I., Velic A., Macek B., Kretschmer D., Weidenmaier C., Hector A., Handgretinger R., Götz F., Peschel A., Hartl D, & Rieber N. (2018). Staphylococcal Enterotoxins Dose-Dependently Modulate the Generation of Myeloid-Derived Suppressor Cells. Frontiers in Cellular and Infection Microbiology, 8, 321.
+ Open protocol
+ Expand
10

Allogeneic Lung MNC Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Allogenic P. alecto lung mononuclear cells (MNC) were labelled with 0.2 μM carboxyfluorescein succinimidyl ester (CFSE) (Thermo Scientific) for 10 min at 37 °C. 10,000 6 day-culture BM-derived DC or macrophages were co-cultured with 100,000 CFSE labelled lung MNC in the presence of 75 UI/ml of recombinant human IL-2 (R&D Systems) for 6 days in AIM-V medium (Thermo Scientific) supplemented with 10% human AB-serum (Sigma Aldrich). As controls, lung MNC were cultured 6 days alone or in the presence of 75 UI/ml of recombinant human IL-2. On day 6, cells were analysed by flow cytometry.
Zhou P., Chionh Y.T., Irac S.E., Ahn M., Jia Ng J.H., Fossum E., Bogen B., Ginhoux F., Irving A.T., Dutertre C.A, & Wang L.F. (2016). Unlocking bat immunology: establishment of Pteropus alecto bone marrow-derived dendritic cells and macrophages. Scientific Reports, 6, 38597.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!