Ctl analyzer

Manufactured by Cellular Technology

Sourced in United States

The CTL Analyzer is a laboratory instrument designed for the analysis of cellular samples. It provides accurate measurements and data on various cellular parameters, enabling researchers and scientists to gain insights into cellular processes and characteristics.

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Lab products found in correlation

Dual color ifn γ il 4 elispot kit, by R&D Systems (1 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Elispot plate, by Merck Group (1 mentions) Rat anti mouse ifnγ capture antibody, by BD (1 mentions) Elispot blue color module, by R&D Systems (1 mentions) 70 mm nylon cell strainer, by BD (1 mentions) Biotin rat anti mouse ifn γ, by BD (1 mentions) Endobulin, by Baxter (1 mentions)

8 protocols using ctl analyzer

Dual-Color IFN-γ/IL-4 ELISpot for Tumor Immunology

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The Dual-Color IFN-γ/IL-4 ELISpot Kit (R&D, USA ) was used to monitor the
primary and cross responses of immunized rats to TSA-treated and irradiated
NuTu-19 cells and tumor cells, respectively. Briefly, the rats were immunized
with TSA-treated and irradiated NuTu-19 cells as described above and sacrificed
2 weeks after the last boost vaccination. The spleen was collected before
splenocyte isolation. Assays using 3×10³ X-ray-irradiated tumor cells
as stimulator cells and 10⁵ recipient splenocytes as responder cells
were carried out. An ELISpot plate reader (CTL Analyzers; Cellular Technology,
USA) was used to automatically enumerate the spots for further analysis.

Liu Y., Yi T., Meng S., Zhao X., Chen X, & Zhang Y. (2024). Trichostatin A-modified vaccine provides superior protection against ovarian cancer formation and development. Brazilian Journal of Medical and Biological Research, 57, e12874.

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IFN-γ ELISPOT Assay for HIV-1 Peptide Responses

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The IFN-γ ELISPOT assay was performed as previously described ^{25 (link)}. Peptides were derived from HIV-1 Gag and Nef consensus subtype B, C and F1, previously described (ref), and for each patient, was used a subtype homologue consensus peptides. Phytohemagglutinin-5μg/mL (Sigma, USA) was used as a positive control, and cells suspended only in culture medium served as a negative control. The spots were counted using an automated ELISPOT reader (CTL Analyzers LLC, Cellular Technology, USA). The results were expressed as spot-forming cells (SFC)/million PBMC. The response was considered positive if ≥ 50 SFC/10⁶ PBMC were detected.

Côrtes F.H., Passaes C.P., Bello G., Teixeira S.L., Vorsatz C., Babic D., Sharkey M., Grinsztejn B., Veloso V., Stevenson M, & Morgado M.G. (2015). HIV controllers with different viral load cut-off levels have distinct virologic and immunologic profiles. Journal of acquired immune deficiency syndromes (1999), 68(4), 377-385.

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Murine Bone Marrow IgG/IgA ELISpot Assay

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ELISpot plates (Millipore, Bedford, MA, United States) were coated with 2 μg/ml ID93 and incubated overnight at 4°C. Plates were washed with PBS, blocked with complete RPMI and 10% FBS for 2 h, at room temperature, and then washed again. Single-cell suspensions from harvested bone marrow were prepared as described above and seeded at 1.5×10⁶ cells per well. The plates were then incubated at 37°C with 5% CO2 for 3 h, washed, and HRP-conjugated anti-mouse IgG (H + L) or IgA antibodies (Southern Biotech, Birmingham, AL, United States) were added for overnight incubation at 4°C. The plates were developed with AEC substrate kits according to the manufacturer’s protocol (Vector Laboratories, Burlingame, CA, United States). Spots were counted using an automated ELISPOT reader (CTL. Analyzer, Cellular Technology Ltd., Shaker Heights, OH, United States). Data were analyzed using ImmunoSpot® software (CTL Analyzer).

Gomez M., Ahmed M., Das S., McCollum J., Mellett L., Swanson R., Gupta A., Carrigy N.B., Wang H., Barona D., Bachchhav S., Gerhardt A., Press C., Archer M.C., Liang H., Seydoux E., Kramer R.M., Kuehl P.J., Vehring R., Khader S.A, & Fox C.B. (2022). Development and Testing of a Spray-Dried Tuberculosis Vaccine Candidate in a Mouse Model. Frontiers in Pharmacology, 12, 799034.

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ELISPOT Assay for T-Cell Responses

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ELISPOT assays were performed as previously described.⁸ Briefly, antigen-pulsed DCs were incubated with autologous T cells at a 1:5 ratio for 40 h. Quantification was performed using a CTL Analyzer (Cellular Technology), and individuals were considered responders if the spot numbers in the triplicate test wells significantly (two-sided Student’s t-test with p < .05 as the responder criterion) exceeded the numbers in the control wells.

Safi S., Yamauchi Y., Stamova S., Rathinasamy A., op den Winkel J., Jünger S., Bucur M., Umansky L., Warth A., Herpel E., Eichhorn M., Winter H., Hoffmann H, & Beckhove P. (2019). Bone marrow expands the repertoire of functional T cells targeting tumor-associated antigens in patients with resectable non-small-cell lung cancer. Oncoimmunology, 8(12), e1671762.

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IFN-γ ELISpot Assay for Murine Splenocytes

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Interferon (IFN)-γ ELISpot assay was performed as described previously [10 (link)]. Briefly, mice splenocytes were isolated from spleens using a 70-mm nylon cell strainer (BD Pharmingen, San Diego, CA, USA) and incubated in media containing 10 ng/mL interleukin-2 for 48 h. Cells were plated in 96-well multiscreen filter plates (Millipore, Billerica, MA, USA) coated with purified rat anti-mouse IFN-γ capture antibody (BD Pharmingen, San Diego, CA, USA) in triplicate at 1 × 10⁶ cells/well and stimulated by H7 HA peptides (GeneOne Life Science, Inc., Seoul, Korea) for 16 h. Biotin rat anti-mouse IFN-γ (BD Pharmingen, San Diego, CA, USA) was used as the detection antibody. Color development was performed according to the manufacturer’s instructions (ELISPOT Blue Color Module, R&D Systems, Minneapolis, MN, USA). The number of spots on the plates was counted using an automated ELISPOT reader system (CTL Analyzer, Cellular Technology, Cleveland, OH, USA).

Choi E.J., Lee H.S., Noh J.Y., Song J.Y., Cheong H.J., Shin O.S., Lee H., Jeong M, & Kim W.J. (2017). Humoral and Cellular Immunogenicity Induced by Avian Influenza A (H7N9) DNA Vaccine in Mice. Infection & Chemotherapy, 49(2), 117-122.

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Quantification of IgG Subclass Secreting Cells

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Enumeration of IgG1, IgG2, IgG3, and IgG4 secreting cells was done using Human IgG1/IgG2/IgG3/IgG4 Four color Fluorospot (Cellular Technology). Low fluorescence PVDF ELISpot plates were coated with anti-human IgG. 100,000 total thymocytes were incubated overnight without stimulation. Bound antibodies were detected with 4-color fluorochrome-conjugated anti-human IgG1, IgG2, IgG3 and IgG4 (Cellular Technology). Spots were analyzed on a CTL Analyzer (Cellular Technology). Detailed Fluorospot counts and analysis are presented in Table S5.

Nuñez S., Moore C., Gao B., Rogers K., Hidalgo Y., del Nido P.J., Restaino S., Naka Y., Bhagat G., Madsen J.C., Bono M.R, & Zorn E. (2016). The human thymus perivascular space is a functional niche for viral-specific plasma cells. Science immunology, 1(6), eaah4447.

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IFN-γ ELISPOT Assay for T Cell Responses

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ELISPOT assays were performed as previously described [47 (link)], with a few modifications. Briefly, antigen-pulsed DCs were incubated with autologous T cells (DC:T cell ratio = 1:5) for 40 h in ELISPOT plates. The number of IFN-γ spot-forming cells was quantified using a CTL Analyzer (Cellular Technology, Cleveland, OH, USA). As a negative control, DCs were loaded with human IgG (Endobulin, Baxter, Unterschleissheim, Germany), which was considered a nonspecific background. Staphylococcal enterotoxin B (SEB) and cytomegalovirus (CMV) were used as positive controls. Individuals were considered responders if the spot numbers in triplicate test wells significantly (two-sided Student´s t-test with p < 0.05 as the responder criterion) exceeded the numbers in control wells.

Safi S., Yamauchi Y., Hoffmann H., Weichert W., Jost P.J., Winter H., Muley T, & Beckhove P. (2020). Circulating Interleukin-4 Is Associated with a Systemic T Cell Response against Tumor-Associated Antigens in Treatment-Naïve Patients with Resectable Non-Small-Cell Lung Cancer. Cancers, 12(12), 3496.

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Evaluating T-cell Responses to HIV Vaccine

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Freshly isolated PBMC, splenocytes and LNMC were resuspended in R10 medium and tested for reactivity to peptide pools based on the vaccine-expressed HIV antigens, using a standard IFN-γ ELISPOT assay, as previously described [32] (link), [33] (link), [34] (link). Two hundred thousand cells per well, in triplicates, were stimulated with pools of synthetic peptides corresponding to subtype C HIV-1 Gag, Pol, Env, Nef and Tat. The peptides were 15-18mers (overlapping by 10 amino acids) and were obtained from the NIH AIDS Research Reagent Program (Bethesda, MD). Spots were analyzed using a Cellular Technology Ltd (CTL) analyzer. A cut-off value for a positive response based on pre-vaccination peptide reactivity was set at 40 SFU/10⁶ cells after subtraction of the background response. Phytohaematoglutinin-M (PHA-M, Sigma-Aldrich) was used as a positive control and a cut-off value of >500 SFU/10⁶ PBMC was used to validate the assay.

Chege G.K., Burgers W.A., Müller T.L., Gray C.M., Shephard E.G., Barnett S.W., Ferrari G., Montefiori D., Williamson C, & Williamson A.L. (2017). DNA-MVA-protein vaccination of rhesus macaques induces HIV-specific immunity in mucosal-associated lymph nodes and functional antibodies. Vaccine, 35(6), 929-937.

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