DCs from patients with T1D (n = 7) were co-cultured with 1 mM PSAB-liposomes (PSAB-DC), 1000 nM Lira (Lira-DC) or combined (PSAB + Lira DC) for 24 h in the presence of 20 μg/ml human insulin (Sigma-Aldrich). DCs were cultured with 20 μg/ml human insulin (Sigma-Aldrich) to obtain immature DCs (iDCs) and adding a cytokine cocktail [1000 IU/ml TNFα and 2000 IU/ml IL-1β (Immunotools) and 1 μM Prostaglandin E2 (Cayman Chemical, Ann Arbor, USA)] to obtain mature DCs (mDC). To assess DCs phenotype, CD25-PE, CD86-FITC, HLA ABC-FITC, HLA DR-FITC, CD14-PE and CD40-APC (Immunotools), CD36-APCCy7, TIM4-APC, CD54-PECy7, TLR2-FITC, CXCR4-APCCy7, CCR2-APC, PD-L1-PECy7, ILT3-PECy7 (Biolegend, San Diego, USA) and CCR7-PECy7 (BD Biosciences) monoclonal antibodies were used to determine their membrane expression. All DCs conditions from the same patient were analysed at the time. All MFI groups were normalized to the MFI of mDCs.
Ccr2 apc
Manufactured by BioLegend
Sourced in United States
CCR2-APC is a fluorophore-conjugated antibody that specifically binds to the CCR2 receptor, which is expressed on the surface of certain immune cells. This product can be used for the identification and analysis of CCR2-positive cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Human insulin, by Merck Group (2 mentions)
Cd86 fitc, by Immunotools (2 mentions)
Hla dr fitc, by Immunotools (2 mentions)
Cd40 apc, by Immunotools (2 mentions)
Cd36 apccy7, by Immunotools (2 mentions)
Tim4 apc, by BioLegend (2 mentions)
Cd54 pecy7, by BioLegend (2 mentions)
Tlr2 fitc, by BioLegend (2 mentions)
Cxcr4 apccy7, by BioLegend (2 mentions)
Prostaglandin e2, by Cayman Chemical (2 mentions)
Cd14 pe, by Immunotools (2 mentions)
Il 1β, by Immunotools (2 mentions)
Hla abc fitc, by Immunotools (2 mentions)
Facs caliber, by BD (2 mentions)
Ccr7 pe cy7, by BD (1 mentions)
Pd l1 pe cy7, by BioLegend (1 mentions)
Cell staining buffer, by BioLegend (1 mentions)
Ccr4 pe, by BioLegend (1 mentions)
Cd45ra apc vio770, by Miltenyi Biotec (1 mentions)
Fitc pe, by BD (1 mentions)
6 protocols using ccr2 apc
1
Immunomodulatory Effects of PSAB-liposomes and Liraglutide on Dendritic Cells in Type 1 Diabetes
2
Characterization of CAR-T Cells by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: CD3 (VioBlue; Miltenyi Biotech or Pacific blue and PE; BioLegend), CD4 (FITC or APC-Cy7; BioLegend), CD8 (PE-Cy7; BioLegend), CD3/CD19 antibodies (FITC/PE; BD), CD28 (PerCP-Cy5.5; eBioscience), PD-1 (FITC; clone: EH12.2H7; BioLegend), TIM-3 (APC-Cy7; BioLegend), LAG-3 (VioBlue; Miltenyi Biotech), CD45RA (APC-Vio770; Miltenyi Biotec or Brilliant Violet; BioLegend), CCR7 (PerCP-Vio770; Miltenyi Biotec or PerCP; BioLegend), CCR2 (APC; Biolegend), CCR4 (PE; Biolegend), CCR5 (Alexa Influenza 488; Biolegend), CXCR2 (PE-Cy7; Biolegend) and CXCR3 (FITC; Biolegend).
Transduction efficacy was determined on day 6 and day 9 of culture by labeling CAR-T cells with biotin-labeled polyclonal goat anti-mouse F(ab)2 antibody (anti-Fab, Jackson Immunoresearch, West Grove, Pennsylvania) and streptavidin (APC conjugated; BioLegend). CD3+F(ab)2+ cells were defined as CAR-T cells. Isotype labeled cells (Jackson Immunoresearch) and untransduced cells served as negative controls. For further characterization, cells were stained with antibodies mentioned above. Cells were washed and re-suspended in cell staining buffer (BioLegend), incubated for 30 min with the antibodies on ice, washed and measured using MACSQuant FACS cytometer (Miltenyi Biotec). Samples were analyzed by FlowJo software (FlowJo LLC, Ashland, Oregon).
Transduction efficacy was determined on day 6 and day 9 of culture by labeling CAR-T cells with biotin-labeled polyclonal goat anti-mouse F(ab)2 antibody (anti-Fab, Jackson Immunoresearch, West Grove, Pennsylvania) and streptavidin (APC conjugated; BioLegend). CD3+F(ab)2+ cells were defined as CAR-T cells. Isotype labeled cells (Jackson Immunoresearch) and untransduced cells served as negative controls. For further characterization, cells were stained with antibodies mentioned above. Cells were washed and re-suspended in cell staining buffer (BioLegend), incubated for 30 min with the antibodies on ice, washed and measured using MACSQuant FACS cytometer (Miltenyi Biotec). Samples were analyzed by FlowJo software (FlowJo LLC, Ashland, Oregon).
3
Immune Cell Phenotyping Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood mononuclear cells, peritoneal cells or lamina propria cells were taken and blocked in 5% fetal bovine serum for 20 min and then stained with CD45-FITC, CD11b-PE, CCR2-APC, Ly6C-PerCP Cy5.5, Ly6G-PerCP Cy5.5, F4/80-APC, or/and CD11c-APC (BioLegend) at 4 °C for 30 min. Data were acquired from FACS caliber (BD Biosciences) and analyzed using FlowJo 7.6 (TreeStar).
4
Cell Surface CCR2 Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were taken and blocked in 5% fetal bovine serum for 20 min and then stained with CCR2-APC (BioLegend) at 4°C for 30 min. Data were acquired from FACScaliber (BD Biosciences) and analyzed using FlowJo 7.6 (TreeStar).
5
Dissecting and Sorting Mouse Lung Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse lung or tumors were dissected and enzymatically digested for 30 min while being shaken at 37°C in a digestion mix containing 0.25% collagenase II (Gibco) and 1 U/ml dispase (Gibco) in PBS. Cell suspensions were filtered through a 100-μm cell strainer followed by washing with 1% FCS in PBS. Antibodies used were as follows: CD31-PE (MCA2388PE; Serotec), CD45-APC (17-0451; eBioscience), CD4-PeCy7 (25-0041; eBioscience), CD8-PerCp-Cy5.5 (45-0081; eBioscience), CD45-FITC (553079; BD), CD19-PE (12-0193; eBioscience), Ly6G-APC (127613; Biolegend), F4/80-APC-efluor 780 (47-4801; eBioscience), CD206-PE (MCA2235PET; AbD Serotec), CD11c-FITC (557400; BD), CCR2-APC (150627; Biolegend). Cells were sorted on an S3e cell sorter (BioRad) or FACSCanto (BD).
6
Characterizing Tolerogenic Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DCs from patients at onset and with established disease were cultured for 24 h with 20 μg/mL human insulin (Sigma-Aldrich, St. Louis, MO, USA) and 1 mmol/L of PSAB-liposomes to determine the effect of insulin chains as autoantigens (tolDCs). As controls, DCs were either cultured with 20 μg/mL human insulin (Sigma-Aldrich) to obtain iDCs or adding a cytokine cocktail —TNF-α (1,000 IU/mL, Immunotools), IL-1β (2,000 IU/mL, Immunotools), and Prostaglandin E2 (PGE2, 1 μmol/L, Cayman Chemical, Ann Arbor, MI, USA)— for 24 h to obtain mature DCs (mDCs). Viability and phenotype were analyzed by flow cytometry (FACSCanto II, BD Biosciences). DCs were stained with 7-AAD (BD Biosciences) and antibodies to CD11c-APC, CD86-FITC, HLA-ABC-FITC, HLA-DR-FITC, CD14-PE and CD40-APC (Immunotools), CD36-APCCy7, TIM4-APC, αvβ5 integrin-PE, CD54-PECy7, TLR2-FITC, CXCR4-APCCy7, CCR2-APC (BioLegend, San Diego, CA, USA). Data were analyzed using FlowJo software (Tree Star Inc, Ashland, OR, USA).
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
Revolutionizing how scientists
search and build protocols!