Log In Sign up
The largest database of trusted experimental protocols

Celltrace violet ctv cell proliferation kit

Manufactured by Thermo Fisher Scientific
Visit Supplier

The CellTrace™ Violet (CTV) Cell Proliferation Kit is a fluorescent cell-labeling solution that can be used to monitor cell division and proliferation in vitro. The kit provides a simple and efficient method for tracking cell division by labeling cells with a violet-fluorescent dye that is equally partitioned between daughter cells during cell division.

Automatically generated - may contain errors

Lab products found in correlation

Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Anti mouse cd16 32 24g2, by BioXCell (2 mentions) Annexin 5 binding buffer, by BioLegend (2 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (2 mentions) Annexin 5 fitc, by BD (2 mentions) Lsm 700 confocal microscope, by Zeiss (2 mentions) Anti cd3 cd28 beads, by Miltenyi Biotec (2 mentions) Celltrace cfse, by Thermo Fisher Scientific (2 mentions) Violet cell proliferation kit, by Thermo Fisher Scientific (2 mentions) Pepinh myd, by InvivoGen (1 mentions) 96 well elisa plate, by Greiner (1 mentions) Goat anti human igm g a f ab 2, by Jackson ImmunoResearch (1 mentions) Anti cd161, by Miltenyi Biotec (1 mentions) Igg2a isotype control, by R&D Systems (1 mentions) Igg1 isotype control, by R&D Systems (1 mentions) Anti cd40, by Miltenyi Biotec (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Il 21, by Miltenyi Biotec (1 mentions) Il 10, by Miltenyi Biotec (1 mentions)

23 protocols using celltrace violet ctv cell proliferation kit

1

Modulation of B Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Isolated B cells were stimulated with oligodeoxynucleotide 2006 (CpG, InvivoGen, Wiltshire, U.K.) for 48 h. TLR signaling inhibition studies used the MyD88 inhibitory peptide Pepinh-MYD at the given concentration (InvivoGen). B cell stimulations were achieved using goat anti-human IgM/G/A F(ab′)2 (Jackson ImmunoResearch) fragments at 1 μg/ml, anti-CD40 (clone S2C6, Macbeth) at 5 μg/ml, or with IL-4 (PeproTech), IL-10, IL-21 (Miltenyi Biotec), PGE2 (Sigma-Aldrich), IL-15 (Miltenyi Biotec), and BAFF (Miltenyi Biotec) (all at 50 ng/ml). Proliferations assays were performed using a CellTrace Violet (CTV) cell proliferation kit (Life Technologies).
The CD161 blocking assay included 2 × 106 B cells/ml in 500 μl in a 48-well plate (Corning) with combinations of CpG (5 μM), anti-CD40 (5 μg/ml) plus IL-4 (50 ng/ml), anti-CD161 at 1 μg/ml (clone 191.B8, Miltenyi Biotec), and IgG2A isotype control (R&D Systems).
For LLT1 crosslinking, recombinant CD161 (R&D Systems) or IgG1 isotype control (R&D Systems) were bound to a 96-well ELISA plate (Greiner Bio-One, Stonehouse, U.K.) overnight prior to the addition of B cells and BCR stimulus as described above.
Llibre A., López-Macías C., Marafioti T., Mehta H., Partridge A., Kanzig C., Rivellese F., Galson J.D., Walker L.J., Milne P., Phillips R.E., Kelly D.F., Freeman G.J., El Shikh M.E., Klenerman P, & Willberg C.B. (2016). LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation. The Journal of Immunology Author Choice, 196(5), 2085-2094.
+ Open protocol
+ Expand
2

Measuring B Cell Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proliferation assays were performed using CellTrace™ Violet (CTV) Cell Proliferation Kit (Life Technologies). Purified B cells were stained with 2.5 μM CTV for 10min at room temperature, following manufacturer's instructions. Cells were then counted and 2 × 105 B cells were plated per well in a U-bottom 96-well plate with CpG (0.5 μg/ml) for 5 days.
Llibre A., López-Macías C., Marafioti T., Mehta H., Partridge A., Kanzig C., Rivellese F., Galson J.D., Walker L.J., Milne P., Phillips R.E., Kelly D.F., Freeman G.J., El Shikh M.E., Klenerman P, & Willberg C.B. (2016). LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation. The Journal of Immunology Author Choice, 196(5), 2085-2094.
+ Open protocol
+ Expand
3

B Cell Proliferation Assay with CD38 Inhibitors

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Isolated PB B cells were labelled using the CellTrace Violet (CTV) Cell Proliferation Kit (Life Technologies), according to manufacturer’s instructions, and cell proliferation was measured by CTV dilution. The cells were cultured in 96-well plates at 1 × 104 cells/well in complete medium (Table S5) and stimulated with 2.5 µg/ml goat α-human IgA/IgG/IgM F(ab′)2 fragments (α-Ig; Jackson ImmunoResearch Laboratories) in combination with 20 ng/ml human recombinant IL-2 (R&D Systems) and 2.5 µg/ml TLR9 agonist, CpG-B ODN 2006 (InvivoGen) for 5/7 d with or without antibodies recognizing CD38: α-CD38 mouse clone HB-7 (Ultra-LEAF Purified; Biolegend) and daratumumab (trade name Darzalex, human, IgG1κ, HuMax-CD38, Genmab/Janssen) both at a concentration of 1 µg/ml (de Weers et al., 2011 (link); Laubach et al., 2014 (link); Matas-Céspedes et al., 2017 (link)). The proliferation was assessed as expansion index. Data were acquired by flow cytometry.
Camponeschi A., Kläsener K., Sundell T., Lundqvist C., Manna P.T., Ayoubzadeh N., Sundqvist M., Thorarinsdottir K., Gatto M., Visentini M., Önnheim K., Aranburu A., Forsman H., Ekwall O., Fogelstrand L., Gjertsson I., Reth M, & Mårtensson I.L. (2022). Human CD38 regulates B cell antigen receptor dynamic organization in normal and malignant B cells. The Journal of Experimental Medicine, 219(9), e20220201.
+ Open protocol
+ Expand
4

Multi-parameter Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies and dilutions are described in the reporting summary. For staining, single cell preparations were made from spleen and lymph node in MACS buffer (PBS with 2% FBS and 2 μM EDTA). Staining panels containing anti-CD93 (Supplementary Fig. 1d) were performed in MACS buffer not supplemented with EDTA. After ACK (Ammonium Chloride) lysis of RBCs, cells were washed once and Fc receptors blocked with anti-mouse CD16/32 (2.4G2, BioXCell). Cells were incubated with antibodies for 45/60 min on ice. Intracellular staining of BCL-6, Foxp3 and MCL-1 were performed using the Foxp3-staining buffer set from eBioscience. The following reagents were used according to the manufacture instructions: PhiPhiLux-G1D2 kit (OncoImmunin Inc.), Annexin V-FITC (BD Biosciences) and Annexin V binding buffer (BioLegend), CellTrace™ Violet (CTV) Cell Proliferation Kit from Life Technologies, LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit and LIVE/DEAD® Fixable Near-IR Dead Cell Stain Kit (Life Technologies). The following gates were applied before the identification of the specific cell types: FSC-A/SSC-A, exclusion of doublets (SSC-H/SSC-W and FSC-H/FSC-W), live cells (negative for Aqua or Near-IR dead stain kit); gating strategies for each population are indicated in each figure legend. Flow cytometry was performed on a LSRII (BD Biosciences) and data analyzed using FlowJo 9.9 software (TreeStar).
Preite S., Cannons J.L., Radtke A.J., Vujkovic-Cvijin I., Gomez-Rodriguez J., Volpi S., Huang B., Cheng J., Collins N., Reilley J., Handon R., Dobbs K., Huq L., Raman I., Zhu C., Li Q.Z., Li M.O., Pittaluga S., Uzel G., Notarangelo L.D., Belkaid Y., Germain R.N, & Schwartzberg P.L. (2018). Hyperactivated PI3Kδ promotes self and commensal reactivity at the expense of optimal humoral immunity. Nature immunology, 19(9), 986-1000.
+ Open protocol
+ Expand
5

Assessing Cellular Morphology with AR Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
To assess morphology, OE33-AR (1.72 × 103), JH-AR (5.73 × 103), or OE19-AR (5.73 × 103) cells were seeded into 96-well µ-plates (Ibidi, Martinsried, Germany) in stripped medium supplemented with either vehicle or 10 nM DHT replaced daily for 3 days. For direct co-cultures, NFFs were labeled using the CellTrace Violet (CTV) Cell Proliferation Kit according to the manufacturer’s protocol (Life Technologies). Fibroblasts were seeded at 1.14 × 104 per well, and OE33-ARs were seeded overnight at 1.43 × 103 cells per well, in either monoculture or overlying fibroblasts followed by treatment with vehicle or 10 nM DHT for 6 days, with medium replaced every 48 h. Images were captured using a Zeiss LSM 700 confocal microscope with Zen2012 SP1 (black edition) software version 8.1.
Palethorpe H.M., Drew P.A, & Smith E. (2017). Androgen Signaling in Esophageal Adenocarcinoma Cell Lines In Vitro. Digestive Diseases and Sciences, 62(12), 3402-3414.
+ Open protocol
+ Expand
6

Cell Labeling for Prostate Cancer

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Luciferase-tagged PC3 and LNCaP cells were generated to express green fluorescent protein (GFP) by stably transducing with the triple reporter gene construct SFG-NES-TGL as described previously [43 (link)] and were a kind gift from Professor Andreas Evdokiou. The C4-2B and DU145 cells were labelled using the CellTrace Violet (CTV) Cell Proliferation Kit according to the manufacturer’s protocol (Life Technologies). The PShTert-AR and PShTert myofibroblasts were stably transduced with the SFG-RFP/Rluc construct to express red fluorescent protein (RFP) as described previously [44 (link)].
Palethorpe H.M., Leach D.A., Need E.F., Drew P.A, & Smith E. (2018). Myofibroblast androgen receptor expression determines cell survival in co-cultures of myofibroblasts and prostate cancer cells in vitro. Oncotarget, 9(27), 19100-19114.
+ Open protocol
+ Expand
7

Multi-parameter Flow Cytometry Assay

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Anti-Bcl-2 PE, CD3-FITC, CD8-APC, CD127-BV510, IFNγ-APC, RORγt-BV421, TNF-PE were from BD Biosciences. Anti-CD4-PerCP, CD8-BV510, CD127-PECy7, CD161-BV421, -FITC, and -PE, IFNγ-APC, Ki-67-AF488 and -BV510, TNF-BV421, and Vα7.2 PECy7 were from Biolegend. Anti-PLZF APC was from R&D Systems. CellTrace Violet (CTV) cell proliferation kit, Live/Dead Aqua and Near Infrared fixable cell stain were from Life Technologies. MR1-5-OP-RU tetramer -APC, -BV421, and -PE were from NIH Tetramer Core Facility, Emory University. Staining with the MR1 5-OP-RU tetramers was performed for 40 min at room temperature (RT) (6 (link)) before proceeding to the surface and intracellular staining with other mAbs. Cell surface and intracellular staining for TFs, cytokines, and cytotoxic molecules were performed as previously described (10 (link)). Samples were acquired on an FACS Canto II (BD Biosciences) equipped with 405, 488, and 633 nm lasers or CytoFLEX (Beckman Coulter) equipped with 405, 488, and 638 nm lasers. Data including the compensation platform were analysed using FlowJo v.10 (BD Biosciences).
Han F., Gulam M.Y., Zheng Y., Zulhaimi N.S., Sia W.R., He D., Ho A., Hadadi L., Liu Z., Qin P., Lobie P.E., Kamarulzaman A., Wang L.F., Sandberg J.K., Lewin S.R., Rajasuriar R, & Leeansyah E. (2022). IL7RA single nucleotide polymorphisms are associated with the size and function of the MAIT cell population in treated HIV-1 infection. Frontiers in Immunology, 13, 985385.
+ Open protocol
+ Expand
8

Multi-parameter Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies and dilutions are described in the reporting summary. For staining, single cell preparations were made from spleen and lymph node in MACS buffer (PBS with 2% FBS and 2 μM EDTA). Staining panels containing anti-CD93 (Supplementary Fig. 1d) were performed in MACS buffer not supplemented with EDTA. After ACK (Ammonium Chloride) lysis of RBCs, cells were washed once and Fc receptors blocked with anti-mouse CD16/32 (2.4G2, BioXCell). Cells were incubated with antibodies for 45/60 min on ice. Intracellular staining of BCL-6, Foxp3 and MCL-1 were performed using the Foxp3-staining buffer set from eBioscience. The following reagents were used according to the manufacture instructions: PhiPhiLux-G1D2 kit (OncoImmunin Inc.), Annexin V-FITC (BD Biosciences) and Annexin V binding buffer (BioLegend), CellTrace™ Violet (CTV) Cell Proliferation Kit from Life Technologies, LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit and LIVE/DEAD® Fixable Near-IR Dead Cell Stain Kit (Life Technologies). The following gates were applied before the identification of the specific cell types: FSC-A/SSC-A, exclusion of doublets (SSC-H/SSC-W and FSC-H/FSC-W), live cells (negative for Aqua or Near-IR dead stain kit); gating strategies for each population are indicated in each figure legend. Flow cytometry was performed on a LSRII (BD Biosciences) and data analyzed using FlowJo 9.9 software (TreeStar).
Preite S., Cannons J.L., Radtke A.J., Vujkovic-Cvijin I., Gomez-Rodriguez J., Volpi S., Huang B., Cheng J., Collins N., Reilley J., Handon R., Dobbs K., Huq L., Raman I., Zhu C., Li Q.Z., Li M.O., Pittaluga S., Uzel G., Notarangelo L.D., Belkaid Y., Germain R.N, & Schwartzberg P.L. (2018). Hyperactivated PI3Kδ promotes self and commensal reactivity at the expense of optimal humoral immunity. Nature immunology, 19(9), 986-1000.
+ Open protocol
+ Expand
9

MAIT Cell Functional Assay with E. coli

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
PBMCs were cultured in 10 ng/ml recombinant human IL-7 (R&D Systems), a combination of IL-12 and IL-18 (10 ng/ml and 100 ng/ml, respectively; PeproTech) for 24–48 h, as indicated or left untreated at 37°C and 5% CO2 in RPMI medium supplemented with 10% fetal calf serum and 50 μg/ml gentamicin (Gibco) (RF10 medium) prior to functional assay. MAIT cell functions were determined in vitro using a paraformaldehyde (PFA)-fixed E. coli stimulation (D21 strain, MOI as indicated) in the presence of 1.25 μg/ml anti-CD28 mAb (BD Biosciences) [11 (link)]. PBMCs were further cultured for 24 h, and in selected experiments, 0.4 μg/ml anti-CD107a PECy7 (BD Biosciences) was added at the start of bacterial stimulation, and monensin (Golgi Stop, BD Biosciences) was added during the last six hours of the stimulation. In selected experiments, cells were stained with Cell Trace Violet (CTV) Cell Proliferation Kit (Life Technologies) as per manufacturer’s instructions and cultured in RF10 medium with fixed E. coli in the presence of anti-CD28 for six days as described [12 (link)].
Leeansyah E., Svärd J., Dias J., Buggert M., Nyström J., Quigley M.F., Moll M., Sönnerborg A., Nowak P, & Sandberg J.K. (2015). Arming of MAIT Cell Cytolytic Antimicrobial Activity Is Induced by IL-7 and Defective in HIV-1 Infection. PLoS Pathogens, 11(8), e1005072.
+ Open protocol
+ Expand
10

T Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
T cells were labeled with a CellTrace Violet (CTV) Cell Proliferation Kit (ThermoFisher Scientific) according to the manufacturer’s instruction. Labeled T cells were co-cultured with target cells at a 1:1 effector-to-target (E:T) ratio for 72 h. Percent proliferating cells and absolute cell counts were analyzed using a MACSQuant Analyzer 10 (Miltenyi Biotec).
Blaeschke F., Ortner E., Stenger D., Mahdawi J., Apfelbeck A., Habjan N., Weißer T., Kaeuferle T., Willier S., Kobold S, & Feuchtinger T. (2022). Design and Evaluation of TIM-3-CD28 Checkpoint Fusion Proteins to Improve Anti-CD19 CAR T-Cell Function. Frontiers in Immunology, 13, 845499.
+ 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!