blood mononuclear cells pbmcs (Millipore)

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Millipore blood mononuclear cells pbmcs

FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in <t>AIH</t> patients. <t>PBMCs</t> were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Blood Mononuclear Cells Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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blood mononuclear cells pbmcs - by Bioz Stars, 2021-03

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Images

1) Product Images from "The Imbalance between Foxp3+Tregs and Th1/Th17/Th22 Cells in Patients with Newly Diagnosed Autoimmune Hepatitis"

Article Title: The Imbalance between Foxp3+Tregs and Th1/Th17/Th22 Cells in Patients with Newly Diagnosed Autoimmune Hepatitis

Journal: Journal of Immunology Research

doi: 10.1155/2018/3753081

FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Figure Legend Snippet: FACS analysis of the numbers of different subsets of circulating effector CD3 + CD4 + T cells and ELISA analysis of serum IFN- γ , IL-17, and IL-22 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with APC-anti-CD4 and PerCP-anti-CD3 or isotype controls, fixed, and permeabilized, followed by intracellular staining with FITC-anti-IL-17 and PE-Cy7-anti-IFN- γ and PE-anti-IL-22. The frequency of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + T cells were calculated, according to the total numbers of PBMCs and different types of CD3 + CD4 + T cells. (a) The flow cytometry analysis; (b–d) the numbers of CD3 + CD4 + IFN- γ + Th1, CD3 + CD4 + IL-17 + Th17, and CD3 + CD4 + IL-22 + Th22 cells; (e–g) serum levels of IFN- γ , IL-17, and IL-22. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Techniques Used: FACS, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Flow Cytometry, Cytometry

FACS analysis of the numbers of different subsets of circulating CD3 + CD4 + T cells and ELISA analysis of serum IL-10 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with FITC-anti-CD3, PE-Cy7-anti-CD25, and PerCP-anti-CD4 or isotype controls, fixed, and permeabilized, followed by intracellular staining with PE-anti-Foxp3. The frequency of CD3 + CD4 + CD25 − Foxp3 + and CD3 + CD4 + CD25 + Foxp3 + T cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + Foxp3 + T cells were calculated, according to the total numbers of PBMCs and the frequency of different types of CD3 + CD4 + Foxp3 + T cells. The concentrations of serum IL-10 in individual subjects were determined by ELISA. (a) Flow cytometry analysis; (b) the numbers of CD3 + CD4 + CD25 + Foxp3 + T cells; (c) the numbers of CD3 + CD 4+ CD25 − Foxp3 + T cells; (d) serum levels of IL-10. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood and the mean levels of serum IL-10 in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Figure Legend Snippet: FACS analysis of the numbers of different subsets of circulating CD3 + CD4 + T cells and ELISA analysis of serum IL-10 in AIH patients. PBMCs were isolated from individual subjects, and PBMCs 5∗10 5 /tube were stained in duplicate with FITC-anti-CD3, PE-Cy7-anti-CD25, and PerCP-anti-CD4 or isotype controls, fixed, and permeabilized, followed by intracellular staining with PE-anti-Foxp3. The frequency of CD3 + CD4 + CD25 − Foxp3 + and CD3 + CD4 + CD25 + Foxp3 + T cells was determined by flow cytometry analysis. The cells were gated on living lymphocytes and then gated on CD3 + CD4 + cells, and at least about 30,000 events were analyzed for each sample. The numbers of each type of CD3 + CD4 + Foxp3 + T cells were calculated, according to the total numbers of PBMCs and the frequency of different types of CD3 + CD4 + Foxp3 + T cells. The concentrations of serum IL-10 in individual subjects were determined by ELISA. (a) Flow cytometry analysis; (b) the numbers of CD3 + CD4 + CD25 + Foxp3 + T cells; (c) the numbers of CD3 + CD 4+ CD25 − Foxp3 + T cells; (d) serum levels of IL-10. Data shown are representative FACS charts or the mean numbers of each type of cells per mL of peripheral blood and the mean levels of serum IL-10 in individual subjects from two separate experiments. The horizontal lines indicate the median values for each group. Data shown are representative charts of different subsets of CD3 + CD4 + T cells and serum IL-10 from individual groups of subjects ( n = 20 for the HC, n = 32 for the patients at 0 week, and n = 19 for the patients at 8 weeks posttreatment).

Techniques Used: FACS, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Flow Cytometry, Cytometry

2) Product Images from "Replication and Meta-Analysis of GWAS Identified Susceptibility Loci in Kawasaki Disease Confirm the Importance of B Lymphoid Tyrosine Kinase (BLK) in Disease Susceptibility"

Article Title: Replication and Meta-Analysis of GWAS Identified Susceptibility Loci in Kawasaki Disease Confirm the Importance of B Lymphoid Tyrosine Kinase (BLK) in Disease Susceptibility

Journal: PLoS ONE

doi: 10.1371/journal.pone.0072037

B cell population in peripheral blood mononuclear cells (PBMC) and BLK expression in peripheral blood leukocytes (PBLs) induced at the acute stage of KD. (A) PBMCs were stained with anti-CD19 or anti-CD3 monoclonal antibodies, and the percentage of CD19 + B cells and CD3 + T cells in samples taken from a patient at acute and convalescent stages of KD and from a healthy control were determined by multicolor flow cytometry. (B) Levels of BLK expression were determined by real-time RT-PCR, and levels in KD patients were compared to those in fever controls. Values are expressed as mean ± standard error (SE). RNA was harvested from PBLs from KD patients at different stages of disease development or from age-matched fever controls. KD1, n = 20, before IVIG treatment (within 24 h before IVIG treatment); KD2, n = 12, after IVIG treatment (3–7 days after IVIG treatment); and KD3, n = 10, convalescence stage (3 weeks after IVIG treatment). FC, n = 19, fever controls.

Figure Legend Snippet: B cell population in peripheral blood mononuclear cells (PBMC) and BLK expression in peripheral blood leukocytes (PBLs) induced at the acute stage of KD. (A) PBMCs were stained with anti-CD19 or anti-CD3 monoclonal antibodies, and the percentage of CD19 + B cells and CD3 + T cells in samples taken from a patient at acute and convalescent stages of KD and from a healthy control were determined by multicolor flow cytometry. (B) Levels of BLK expression were determined by real-time RT-PCR, and levels in KD patients were compared to those in fever controls. Values are expressed as mean ± standard error (SE). RNA was harvested from PBLs from KD patients at different stages of disease development or from age-matched fever controls. KD1, n = 20, before IVIG treatment (within 24 h before IVIG treatment); KD2, n = 12, after IVIG treatment (3–7 days after IVIG treatment); and KD3, n = 10, convalescence stage (3 weeks after IVIG treatment). FC, n = 19, fever controls.

Techniques Used: Expressing, Staining, Flow Cytometry, Cytometry, Quantitative RT-PCR

3) Product Images from "Long-Lasting Enfuvirtide Carrier Pentasaccharide Conjugates with Potent Anti-Human Immunodeficiency Virus Type 1 Activity ▿"

Article Title: Long-Lasting Enfuvirtide Carrier Pentasaccharide Conjugates with Potent Anti-Human Immunodeficiency Virus Type 1 Activity ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00827-09

Antiviral activities (EC 50 s) of enfuvirtide-CP conjugates EP40111 and EP40112 against HIV-1 IIIB/LAI infecting PBMCs in the absence or the presence of 2.5 μM antithrombin. These results represent the means and standard deviations of three independent

Figure Legend Snippet: Antiviral activities (EC 50 s) of enfuvirtide-CP conjugates EP40111 and EP40112 against HIV-1 IIIB/LAI infecting PBMCs in the absence or the presence of 2.5 μM antithrombin. These results represent the means and standard deviations of three independent

Techniques Used:

4) Product Images from "Hepatocellular carcinoma cell sensitivity to Vγ9Vδ2 T lymphocyte-mediated killing is increased by zoledronate"

Article Title: Hepatocellular carcinoma cell sensitivity to Vγ9Vδ2 T lymphocyte-mediated killing is increased by zoledronate

Journal: International Journal of Oncology

doi: 10.3892/ijo.2016.3403

γδ T cell effects on Zol-treated HCC cell lines. (A) Maturation stage of γδ T cells, indicated by CD27 and CD45RA expression. Upper, representative plot of CD3 + TCRVγ9 + γδ T cells gated from healthy donor PBMCs. Lower, maturation of CD3 + TCRVγ9 + γδ T cells in 14-day cultures stimulated with PHA in the presence of IL-2 and IL-15. (B) Cytotoxic activity of γδ T cells towards target cells preincubated for 16 h in the presence or absence of 5 μM Zol. γδ T effector (E) cells were co-cultured for 4 h with indicated target (T) cells (1.0×10 4 cells/well) at the indicated E:T ratios. EJ1 cells, T2 cells, and PHA blast served as references. Data shown are representative of three independent experiments. (C) Cytokine production by γδ T cells (1.0×10 5 cells/well) after 24-h co-culture with the indicated target cells (5.0×10 4 cells/well). Data represent mean ± SD of triplicate cultures. (D) Proliferative response of γδ T cells (5.0×10 4 cells/well) co-cultured with irradiated (90 Gy) HCC cell lines (5.0×10 3 cells/well) for 72 h, as determined by [ 3 H]-thymidine incorporation assay. Data represent mean ± SD of triplicate cultures. Significance was determined by a one-way ANOVA with the Bonferroni post-hoc test; * P

Figure Legend Snippet: γδ T cell effects on Zol-treated HCC cell lines. (A) Maturation stage of γδ T cells, indicated by CD27 and CD45RA expression. Upper, representative plot of CD3 + TCRVγ9 + γδ T cells gated from healthy donor PBMCs. Lower, maturation of CD3 + TCRVγ9 + γδ T cells in 14-day cultures stimulated with PHA in the presence of IL-2 and IL-15. (B) Cytotoxic activity of γδ T cells towards target cells preincubated for 16 h in the presence or absence of 5 μM Zol. γδ T effector (E) cells were co-cultured for 4 h with indicated target (T) cells (1.0×10 4 cells/well) at the indicated E:T ratios. EJ1 cells, T2 cells, and PHA blast served as references. Data shown are representative of three independent experiments. (C) Cytokine production by γδ T cells (1.0×10 5 cells/well) after 24-h co-culture with the indicated target cells (5.0×10 4 cells/well). Data represent mean ± SD of triplicate cultures. (D) Proliferative response of γδ T cells (5.0×10 4 cells/well) co-cultured with irradiated (90 Gy) HCC cell lines (5.0×10 3 cells/well) for 72 h, as determined by [ 3 H]-thymidine incorporation assay. Data represent mean ± SD of triplicate cultures. Significance was determined by a one-way ANOVA with the Bonferroni post-hoc test; * P

Techniques Used: Expressing, Activity Assay, Cell Culture, Co-Culture Assay, Irradiation, Thymidine Incorporation Assay

other:

Article Title: Role of Bak in UV-induced apoptosis in skin cancer and abrogation by HPV E6 proteins
Article Snippet: The monoclonal antibodies used were Bak (Ab-2, Calbiochem), p53 (D01, ICRF), p21 (Santa Cruz), Ki67 (Dako), α-Tubulin (Calbiochem), and anti-HA (12CA5, ICRF).

Incubation:

Article Title: SPARCL1 suppresses metastasis in prostate cancer
Article Snippet: .. Membranes were incubated with chicken polyclonal antibody specific for human SPARCL1 (Abcam, Cambridge, United Kingdom) at a 1:2000 dilution, and then the membranes were re‐probed using mouse monoclonal antibody specific for human anti‐β‐actin (Sigma Chemical Company, Saint Louis, MO, USA). .. The human gene SPARCL1 ORF (RC207583, OriGene Technologies, Inc, Rockville, MD) was subcloned into pBMN‐I‐GFP (Addgene Inc., Cambridge, MA, USA) to obtain a pBMN‐SPARCL1‐I‐GFP plasmid.

Article Title: Conditional Site-Specific Integration into Human Chromosome 19 by Using a Ligand-Dependent Chimeric Adeno-Associated Virus/Rep Protein
Article Snippet: The polyclonal antiserum against Rep proteins was obtained by immunizing rabbits with purified recombinant Rep68 produced in Escherichia coli ( ) and recognizes all four species of Rep. To check the expression of the Hook gene product (sFv/PDGFR fusion protein) ( ) in the stable transfectants, total cellular extracts were prepared as described above, fractionated on an SDS–12% polyacrylamide gel, and transferred to nitrocellulose membranes. .. To detect the sFv/PDGFR fusion protein, the membranes were incubated first with monoclonal antibody 9E10.2 (dilution, 1:500), which recognizes the Myc.1 epitope tag ( ) present as a tandem repeat near the transmembrane domain of sFv/PDGFR , and then with an alkaline phosphatase-conjugated goat polyclonal anti-mouse IgG antiserum (Sigma no. A7434; dilution, 1:2,000). .. In all immunoblotting experiments, 5% nonfat dry milk in TBST (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 0.05% Tween 20) was used as a blocking agent and for diluting the various antibodies.

Inhibition:

Article Title: The Roles of Integrins in Function of Human Neutrophils after Their Migration through Endothelium into Interstitial Matrix
Article Snippet: The particles have a low fluorescence intensity in standard media or when adhered to the surface of neutrophils, but increase their fluorescence when taken into acidic phagocytic vacuoles. .. Inhibition of integrin function The following antibodies were used at 10μg/ml to block integrin function: rat IgG2a anti-human CD29/β1-integrin (Mab13; BD Pharmingen); mouse IgG2a anti-human CD18/β2-integrin (IB4; Calbiochem); mouse IgG1 anti-human CD61/β3-integrin (SZ21; Beckman Coulter); matched control antibodies, rat IgG2a, mouse IgG2a (both eBioscience), mouse IgG1 (DAKO). .. In some experiments, RGDS peptide (Arg-Gly-Asp-Ser, 0.5 mM; Sigma) or CT7010 (20 μM) was used instead of antibody.