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human anti leptin neutralizing mab (R&D Systems)

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R&D Systems human anti leptin neutralizing mab
Human Anti Leptin Neutralizing Mab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+anti+leptin+neutralizing+mab/Human+Leptin%2FOB+Antibody/pmc04760097__mmc2-231-8-16
Average 93 stars, based on 21 article reviews

human anti leptin neutralizing mab - by Bioz Stars, 2026-10

93/100 stars

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In Vitro:

Article Title: The Proteomic Landscape of Human Ex Vivo Regulatory and Conventional T Cells Reveals Specific Metabolic Requirements
Article Snippet: Cells were stimulated for 3 days in the presence of anti-CD3 and anti-CD28-coated Dynabeads (0.1 beads per cell) (Invitrogen). .. For in vitro leptin blocking experiments, we used human anti-leptin neutralizing mAb (clone 44802, Cat.# MAB398, R&D Systems) at a final concentration of 20 mg/ml. .. 2-Deoxy-D-glucose (2DG) (Sigma-Aldrich) was used at final concentration of 1 mM; etomoxir was purchased from Sigma-Aldrich and it was used at the final concentration of 200 mM.

Article Title: The Proteomic Landscape of Human Ex Vivo Regulatory and Conventional T Cells Reveals Specific Metabolic Requirements
Article Snippet: Cells were stimulated for 3 days in the presence of anti-CD3 and anti-CD28-coated Dynabeads (0.1 beads per cell) (Invitrogen). .. For in vitro leptin blocking experiments, we used human anti-leptin neutralizing mAb (clone 44802, Cat.# MAB398, R&D Systems) at a final concentration of 20 μg/ml. .. 2-Deoxy-D-glucose (2DG) (Sigma-Aldrich) was used at final concentration of 1 mM; etomoxir was purchased from Sigma-Aldrich and it was used at the final concentration of 200 μM.

Article Title: Regulatory T cell proliferative potential is impaired in human autoimmune disease.
Article Snippet: Human CD4+CD25highCD127−FoxP3+ regulatory T (Treg) cells suppress immune responses in vitro and in vivo1.. Reduced suppressive function and/or number of peripheral Treg cells has been previously reported in autoimmune disorders2,3.. Treg cells represent the most actively replicating compartment within the CD4+ cells in vivo, but they are hyporesponsive to classical T cell receptor (TCR) stimulation in vitro, a condition that is secondary to their overactive metabolic state4,5.

Blocking Assay:

Article Title: The Proteomic Landscape of Human Ex Vivo Regulatory and Conventional T Cells Reveals Specific Metabolic Requirements
Article Snippet: Cells were stimulated for 3 days in the presence of anti-CD3 and anti-CD28-coated Dynabeads (0.1 beads per cell) (Invitrogen). .. For in vitro leptin blocking experiments, we used human anti-leptin neutralizing mAb (clone 44802, Cat.# MAB398, R&D Systems) at a final concentration of 20 μg/ml. .. 2-Deoxy-D-glucose (2DG) (Sigma-Aldrich) was used at final concentration of 1 mM; etomoxir was purchased from Sigma-Aldrich and it was used at the final concentration of 200 μM.

Concentration Assay:

Article Title: The Proteomic Landscape of Human Ex Vivo Regulatory and Conventional T Cells Reveals Specific Metabolic Requirements
Article Snippet: Cells were stimulated for 3 days in the presence of anti-CD3 and anti-CD28-coated Dynabeads (0.1 beads per cell) (Invitrogen). .. For in vitro leptin blocking experiments, we used human anti-leptin neutralizing mAb (clone 44802, Cat.# MAB398, R&D Systems) at a final concentration of 20 μg/ml. .. 2-Deoxy-D-glucose (2DG) (Sigma-Aldrich) was used at final concentration of 1 mM; etomoxir was purchased from Sigma-Aldrich and it was used at the final concentration of 200 μM.

Recombinant:

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Figure 1. Effects of leptin on viability on eosinophils. Eosinophils (5 105/well) were cultured with or without leptin (50 ng/mL) for 24 h in a 24-well plate. Apoptosis of eosinophils was assessed by the TACSTM Annexin V-FITC assay using flow cytometry. Representative dot plots of (A) medium control (CTL), and (B) leptin-treated eosinophils were shown from triplicate experiments.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 1. Effects of leptin on viability on eosinophils. Eosinophils (5 105/well) were cultured with or without leptin (50 ng/mL) for 24 h in a 24-well plate. Apoptosis of eosinophils was assessed by the TACSTM Annexin V-FITC assay using flow cytometry. Representative dot plots of (A) medium control (CTL), and (B) leptin-treated eosinophils were shown from triplicate experiments.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Cell Culture, Flow Cytometry, Control

Figure 2. Effects of leptin on cell surface expression of ICAM-1, CD18, ICAM-3 and L-selectin on eosinophils. Eosinophils (5 105/well) were cultured with leptin (0–100 ng/mL) for 16 h in a 24-well plate. Surface expression of adhesion molecules per 10 000 viable cells was analyzed by flow cytometry as MFI; (A) representative histograms of the changes in the expression of adhesion molecules. Results of (B) ICAM-1, (C) CD18, (D) ICAM-3 and (E) L-selectin have been normalized by subtracting appropriate isotypic control and are expressed as the arithmetic mean SD of five independent experiments. * p<0.05, ** p<0.01 and *** p<0.001 when compared with medium control.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 2. Effects of leptin on cell surface expression of ICAM-1, CD18, ICAM-3 and L-selectin on eosinophils. Eosinophils (5 105/well) were cultured with leptin (0–100 ng/mL) for 16 h in a 24-well plate. Surface expression of adhesion molecules per 10 000 viable cells was analyzed by flow cytometry as MFI; (A) representative histograms of the changes in the expression of adhesion molecules. Results of (B) ICAM-1, (C) CD18, (D) ICAM-3 and (E) L-selectin have been normalized by subtracting appropriate isotypic control and are expressed as the arithmetic mean SD of five independent experiments. * p<0.05, ** p<0.01 and *** p<0.001 when compared with medium control.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Expressing, Cell Culture, Flow Cytometry, Control

Fig. 10 shows that leptin-induced release of IL-1b and IL-6, and IL-8, GRO-a and MCP-1 was suppressed by BAY11–7082 and SB203580, and BAY11–7082, PD98059 and SB203580, respectively (all p<0.05).

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Fig. 10 shows that leptin-induced release of IL-1b and IL-6, and IL-8, GRO-a and MCP-1 was suppressed by BAY11–7082 and SB203580, and BAY11–7082, PD98059 and SB203580, respectively (all p<0.05).

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques:

Figure 4. Effects of neutralization antibody against leptin on the biological activities of leptin on eosinophils. Leptin was incubated with or without neutralization antibody (0.1–10 lg/mL) for 15 min and then incubated with eosinophils (5 105/well) for the assay of (A) apoptosis, (B) cell surface of adhesion molecules, (C) chemokinesis and (D) induction of cytokines. Results were expressed as arithmetic mean plus SD from triplicate experiments. Mouse IgG1 was used as a nonspecific antibody for isotypic antibody control. # p<0.05; ## p<0.01 when compared with medium control (CTL); * p<0.05, ** p<0.01, *** p<0.001 when compared with leptin control (lep). Ab0.1: neutralization antibody (0.1 lg/mL); Ab1: neutralization antibody (1 lg/mL); Ab10: neutralization antibody (10 lg/mL); mIgG: mouse IgG1 (10 lg/mL).

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 4. Effects of neutralization antibody against leptin on the biological activities of leptin on eosinophils. Leptin was incubated with or without neutralization antibody (0.1–10 lg/mL) for 15 min and then incubated with eosinophils (5 105/well) for the assay of (A) apoptosis, (B) cell surface of adhesion molecules, (C) chemokinesis and (D) induction of cytokines. Results were expressed as arithmetic mean plus SD from triplicate experiments. Mouse IgG1 was used as a nonspecific antibody for isotypic antibody control. # p<0.05; ## p<0.01 when compared with medium control (CTL); * p<0.05, ** p<0.01, *** p<0.001 when compared with leptin control (lep). Ab0.1: neutralization antibody (0.1 lg/mL); Ab1: neutralization antibody (1 lg/mL); Ab10: neutralization antibody (10 lg/mL); mIgG: mouse IgG1 (10 lg/mL).

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Neutralization, Incubation, Control

Figure 5. Representative expression profiles of intracellular phosphorylation of MAPK and other serine/threonine kinases of eosinophils upon activation by leptin. Eosinophils (1 107/ well) were treated with or without leptin (50 ng/mL) for 15 min. Eosinophils were then washed and lysed. Twenty-one different intracellular phosphorylated MAPK and other serine/threonine kinases in total cellular lysate were assessed semi-quantita- tively using an antibody based Human Phospho-MAPK Array Kit. Positive and negative controls were designated at (A1, A2, A21, A22, F1, F2) and (E3–E14), respectively. Triplicate experi- ments were performed with essentially identical results. The format of the antibodies on the array are detailed in Table 2.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 5. Representative expression profiles of intracellular phosphorylation of MAPK and other serine/threonine kinases of eosinophils upon activation by leptin. Eosinophils (1 107/ well) were treated with or without leptin (50 ng/mL) for 15 min. Eosinophils were then washed and lysed. Twenty-one different intracellular phosphorylated MAPK and other serine/threonine kinases in total cellular lysate were assessed semi-quantita- tively using an antibody based Human Phospho-MAPK Array Kit. Positive and negative controls were designated at (A1, A2, A21, A22, F1, F2) and (E3–E14), respectively. Triplicate experi- ments were performed with essentially identical results. The format of the antibodies on the array are detailed in Table 2.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Expressing, Phospho-proteomics, Activation Assay

Figure 6. Activation of ERK, p38 MAPK, JAK2 and NF-jB activities in eosinophils by leptin. Eosinophils (1 107/well) were cultured with or without leptin (50 ng/mL) for different indicated incubation times. Total cellular proteins were extracted from eosinophils for the measurement of (A) phosphorylated and total p38 MAPK, (B) phosphorylated and total ERK, (C) phosphorylated and total IjBa, and (D) phosphorylated and total JAK2 by Western blot analysis. Experiments were performed in three independent replicates with essentially identical results, and representative results are shown.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 6. Activation of ERK, p38 MAPK, JAK2 and NF-jB activities in eosinophils by leptin. Eosinophils (1 107/well) were cultured with or without leptin (50 ng/mL) for different indicated incubation times. Total cellular proteins were extracted from eosinophils for the measurement of (A) phosphorylated and total p38 MAPK, (B) phosphorylated and total ERK, (C) phosphorylated and total IjBa, and (D) phosphorylated and total JAK2 by Western blot analysis. Experiments were performed in three independent replicates with essentially identical results, and representative results are shown.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Activation Assay, Cell Culture, Incubation, Western Blot

Figure 7. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on viability of leptin-treated eosino- phils. Eosinophils (5 105/well) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for 24 h in a 24- well plate. Apoptosis of eosinophils was assessed by the TACSTM Annexin V-FITC assay using flow cytometry. Results were expressed as arithmetic mean plus SD from triplicate experiments. ## p<0.01 when compared with medium control; * p<0.05 when compared with the leptin control. AG: AG490; BAY: BAY11–7082; PD: PD98059; LY: LY294002; SB: SB203580; SP: SP600125; DM: DMSO-

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 7. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on viability of leptin-treated eosino- phils. Eosinophils (5 105/well) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for 24 h in a 24- well plate. Apoptosis of eosinophils was assessed by the TACSTM Annexin V-FITC assay using flow cytometry. Results were expressed as arithmetic mean plus SD from triplicate experiments. ## p<0.01 when compared with medium control; * p<0.05 when compared with the leptin control. AG: AG490; BAY: BAY11–7082; PD: PD98059; LY: LY294002; SB: SB203580; SP: SP600125; DM: DMSO-

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Incubation, Flow Cytometry, Control

Figure 8. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on leptin-induced cell surface expression of (A) ICAM-1, (B) ICAM-3, (C) CD18, and (D) L-selectin on eosinophils. Eosinophils (5 105/well) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for further 16 h. Surface expression of the adhesion molecules of 10 000 cells was assessed by flow cytometry as MFI. Results are expressed as the arithmetic mean plus SD from five independent experiments. DMSO (0.1%) was used as the DMSO control. ### p<0.001 when compared with medium control; * p<0.05, ** p<0.01 when compared with the leptin control.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 8. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on leptin-induced cell surface expression of (A) ICAM-1, (B) ICAM-3, (C) CD18, and (D) L-selectin on eosinophils. Eosinophils (5 105/well) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for further 16 h. Surface expression of the adhesion molecules of 10 000 cells was assessed by flow cytometry as MFI. Results are expressed as the arithmetic mean plus SD from five independent experiments. DMSO (0.1%) was used as the DMSO control. ### p<0.001 when compared with medium control; * p<0.05, ** p<0.01 when compared with the leptin control.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Expressing, Incubation, Flow Cytometry, Control

Figure 9. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on the migration of eosinophils induced by leptin. Eosinophils (5 104 cells) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for further 16 h. Eosinophils were then incubated in a 48-well microchamber in the upper chambers, while the RPMI medium was placed in the lower chambers. Migration was carried out for 6 h. Results are expressed as cell number/high power field with arithmetic mean plus SD of three independent experi- ments. # p<0.05 when compared with medium control; * p<0.05 when compared with the leptin control.

Journal: European journal of immunology

Article Title: Leptin-mediated cytokine release and migration of eosinophils: implications for immunopathophysiology of allergic inflammation.

doi: 10.1002/eji.200636866

Figure Lengend Snippet: Figure 9. Effects of AG490, BAY11–7082, LY294002, PD98059, SB203580 and SP600125 on the migration of eosinophils induced by leptin. Eosinophils (5 104 cells) were pretreated with AG490 (10 lM), BAY11–7082 (1 lM), PD98059 (10 lM), LY294002 (5 lM), SB203580 (7.5 lM) or SP600125 (3 lM) for 1 h followed by incubation with or without leptin (50 ng/mL) for further 16 h. Eosinophils were then incubated in a 48-well microchamber in the upper chambers, while the RPMI medium was placed in the lower chambers. Migration was carried out for 6 h. Results are expressed as cell number/high power field with arithmetic mean plus SD of three independent experi- ments. # p<0.05 when compared with medium control; * p<0.05 when compared with the leptin control.

Article Snippet: Anti-human leptin neutralization mAb was from R&D Systems (MN, USA) and mouse IgG1 antibody was from BD PharMingen (CA, USA).

Techniques: Migration, Incubation, Control

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human anti leptin neutralizing mab (R&D Systems)

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