Ascites and blood were collected at predefined time points, and cells were obtained by centrifugation. After washing or red blood cell lysis, the harvested cells were stained with anti-CD3-allophycocyanin (APC) (17-0032082, eBioscience, USA), anti-CD4-fluorescein isothiocyanate (FITC) (557307, BD Biosciences, USA), anti-CD8a-peridinin chlorophyll protein (PerCP)-Cy5.5 (45-0081-82, eBioscience, USA), or anti-NK1.1-FITC (553164, BD Biosciences, USA). The fluorescence intensity of the cells was then detected using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA).
Anti cd4 fluorescein isothiocyanate fitc
Manufactured by BD
Sourced in United States
Anti-CD4-fluorescein isothiocyanate (FITC) is a fluorescently labeled antibody that binds to the CD4 surface marker. It is designed for use in flow cytometry applications to identify and analyze CD4-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (2 mentions)
Anti pd 1 pe, by BD (2 mentions)
Anti nk1.1 fitc, by BD (1 mentions)
Facscanto 2 cytometer, by BD (1 mentions)
Isotype matched control antibody, by BD (1 mentions)
Anti cd40l pe, by Thermo Fisher Scientific (1 mentions)
Anti icos pe, by BD (1 mentions)
Anti il 21r pe, by BD (1 mentions)
Anti cd161 pe, by BD (1 mentions)
Anti ifn γ fitc, by BD (1 mentions)
Anti ccr6 pe, by BD (1 mentions)
Anti interferon ifn γ fitc, by BD (1 mentions)
Anti il 17a apc, by BD (1 mentions)
Anti cd4 percp, by BD (1 mentions)
Anti cd45ro pe, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
Anti cd4 phycoerythrin, by BD (1 mentions)
Anti cd8 fitc, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Lsrfortessa cell analyzer, by BD (1 mentions)
6 protocols using anti cd4 fluorescein isothiocyanate fitc
1
Immune Cell Phenotyping Protocol
2
Analyzing T Cell Subsets in Chronic HBV
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Five milliliters of heparinized peripheral venous blood was obtained from either healthy volunteers or patients with chronic HBV infection. After removing plasma, the red blood cells were lysed using an NH4Cl lysis solution. Flow cytometry analysis was performed on 106 cells per tube using the following fluorochrome-conjugated antibodies: anti-CD3–phycoerythrin (PE)–cyanine (CY) 7 (eBioscience, San Diego, CA, USA), anti- CD4–fluorescein isothiocyanate (FITC) (BD Company, San Jose, CA, USA), anti- CXCR5–allophycocyanin (APC) (BD Company, San Jose, CA, USA), anti-ICOS–PE (BD Company, San Jose, CA, USA), anti-PD1-PE (BD Company, San Jose, CA, USA), anti-CD40L–PE (eBioscience, San Diego, CA, USA), and anti-IL-21R-PE (BD Company, San Jose, CA, USA). Isotype-matched control antibodies (Beckton Dickinson, San Jose, USA) were used to correct nonspecific binding. After staining for 30 min at 4°C, the cells were washed twice with PBS containing 0.5% bovine serum albumin and subsequently analyzed using a FACS Canto II cytometer and FACSDiva software, version 4.1 (Becton Dickinson).
3
Multiparameter Flow Cytometry Analysis
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The following anti-human monoclonal antibodies were used for surface phenotype and intracellular cytokine staining: anti-CD4-fluorescein isothiocyanate (FITC); anti-CD4-phycoerythrin (PE); anti-CD161-PE; anti-CCR6-PE; anti-CD45RO-PE; anti-CD147-peridinin chlorophylla protein cyanine 5.5 dies (Percp–cy5.5); anti-interferon (IFN)-γ-FITC (all from BD Biosciences); and anti-IL-17A-allophycocyanin (APC; eBiosciences). Anti-mouse monoclonal antibodies included: anti-CD4-Percp; anti-IL-17A-APC; and anti-IFN-γ-FITC (all from BD Biosciences). Appropriately conjugated IgG antibodies were used as isotype controls. Cells were acquired on a FACSCalibur flow cytometer (BD Biosciences) and analyzed using Cell Quest software (BD Bioscience) and FlowJo 7.6.1 software (Tree Star).
4
Lymphocyte Profiling with Costimulatory Agents
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Lymphocytes were harvested from the mice spleens and seeded into 24-well plates at 1×106 cells per well before incubation with PS, Cub, and Cub-PS for 48 hours. The cells were then stained with antibodies, including anti-CD3e-phycoerythrin-cyanine5 (anti-CD3e-PE-Cy5), anti-CD8-PE, and anti-CD4-fluorescein isothiocyanate (FITC) antibodies (all from BD Biosciences, San Jose, CA, USA). The cells were analyzed using fluorescence-activated cell sorting analysis.
5
Gastric Adenocarcinoma Immune Profiling
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Patients and healthy donors. Seventy-two patients (53 male and 19 female) with gastric adenocarcinoma who underwent curative gastrectomy (R0 resection) at our Institution between April 2009 and April 2013 were enrolled in this study. The study protocol was approved by the Institutional Review Board of Tottori University Hospital (Yonago, Japan; Approval number 448). None of the patients received radiotherapy, chemotherapy, or other medical interventions before surgery. Clinicopathological findings were determined according to the Japanese Classification of Gastric Carcinoma (16) .
Peripheral blood mononuclear cells (PBMC) preparation. A 30-ml peripheral blood sample was drawn from each of the controls or patients before surgery and centrifuged through a Ficoll-Paque gradient (Pharmacia, Uppsala, Sweden). Informed consent was obtained from all participants before blood donation.
Flow cytometric analysis. Fluorescence-activated cell sorting (FACS) analysis was performed using a FACSCalibur™ (BD Biosciences, Franklin Lakes, NJ, USA). The following antibodies were used to classify cells: anti-CD3-phycoerythrin (PE)-cyanine 5 (Biolegend, San Diego, CA, USA), anti-CD4-fluorescein isothiocyanate (FITC), anti-CD8-FITC, and anti-PD1-PE (all from BD Biosciences).
Peripheral blood mononuclear cells (PBMC) preparation. A 30-ml peripheral blood sample was drawn from each of the controls or patients before surgery and centrifuged through a Ficoll-Paque gradient (Pharmacia, Uppsala, Sweden). Informed consent was obtained from all participants before blood donation.
Flow cytometric analysis. Fluorescence-activated cell sorting (FACS) analysis was performed using a FACSCalibur™ (BD Biosciences, Franklin Lakes, NJ, USA). The following antibodies were used to classify cells: anti-CD3-phycoerythrin (PE)-cyanine 5 (Biolegend, San Diego, CA, USA), anti-CD4-fluorescein isothiocyanate (FITC), anti-CD8-FITC, and anti-PD1-PE (all from BD Biosciences).
6
Endogenous CD4 T cell responses and B cell costimulatory molecules in infected chimera
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Endogenous CD4 T cell responses in infected chimera were analyzed as described previously (Nothelfer et al., 2015) . Briefly, splenocytes were either restimulated with bone marrow-derived dendritic cells (BMDCs) pulsed with fixed parasites or phorbol 12-myristate 13-acetate (PMA)/ionomycin in the presence of Brefeldin A (BD Biosciences). Cells were then stained with anti-CD4-fluorescein isothiocyanate (FITC), anti-CD8 Pacific Blue, anti-IFN-g-allophycocyanin, and anti-IL-10-phycoerythrin (BD Biosciences). 350,000 cells were acquired on a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
The expression of costimulatory molecules by B cells from infected chimeric mice was assessed using the following antibodies: FITC-conjugated anti-major histocompatibility complex class II (MHCII) (BD Biosciences), eFluor 450-conjugated anti-CD19 (eBioscience), PE-conjugated anti-CD40 (eBioscience), PE-conjugated anti-CD80 (eBioscience), and PE-conjugated anti-CD86 (eBioscience). Cells were acquired with a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
The expression of costimulatory molecules by B cells from infected chimeric mice was assessed using the following antibodies: FITC-conjugated anti-major histocompatibility complex class II (MHCII) (BD Biosciences), eFluor 450-conjugated anti-CD19 (eBioscience), PE-conjugated anti-CD40 (eBioscience), PE-conjugated anti-CD80 (eBioscience), and PE-conjugated anti-CD86 (eBioscience). Cells were acquired with a BD LSRFortessa cell analyzer (Becton Dickinson), and analysis was performed using FlowJo software (Tree Star).
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