polyclonal rabbit antibody against gpr44 hpa014259  (Atlas Antibodies)

Journal: EJNMMI Research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: In vitro ligand binding to membranes of HEK293 cells transfected with human recombinant GPR44 and ligand potency at human GPR44 using a clonal beta cell line (EndoC-βH1). [ 3 H]ProstaglandinD 2 ( a ) and [6- 3 H-phenoxy]-AZ12204657 ( b ) binding to human GPR44 was displaced with increasing concentrations of AZ12204657. Results are from two independent experiments with three measurements at each concentration and presented as mean ± SD. The potency of AZ12204657 to inhibit the signal of 15(R)-15-methyl-PGD 2 in human beta cells measured by the label free DMR assay ( c ). Results are from two independent experiments with two measurements at each concentration and presented as mean ± SD

Article Snippet: Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa-647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: In Vitro, Ligand Binding Assay, Transfection, Recombinant, Binding Assay, Concentration Assay

Journal: EJNMMI Research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: In vitro autoradiography of [ 11 C]AZ12204657 reveals focal binding patterns on pancreatic sections from non-diabetic human donors ( a ). The binding could be competed away with an excess of GPR44 antagonist AZD3825 ( b ). Similar GPR44-mediated binding was seen in pancreatic sections from T2D donors ( c , d ). The focal binding corresponded to Islets of Langerhans as assessed by immunofluorescent insulin staining ( e , f ) (representative results from three independent experiments on human sections with measurements performed in duplicates). The islet specific targeting was further assessed by [ 11 C]AZ12204657 binding to homogenates of purified human islets of Langerhans and exocrine tissue preparations (results are from two independent experiments on homogenates with measurements performed in triplicates) ( g ). Binding of [ 11 C]AZ12204657 in pancreatic sections from NHP was similarly focal in nature ( h ), consistent with the heterogeneous distribution of islets of Langerhans ( i ), and GPR44-mediated ( j ) (representative results from six independent experiments on NHP sections with measurements performed in singlets or duplicates)

Article Snippet: Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa-647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: In Vitro, Autoradiography, Binding Assay, Staining, Purification

Journal: EJNMMI Research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: Confocal microscopy of a human islet, showing antibody staining for cell nucleus (blue, a ), insulin (green, b ), GPR44 (red, c ) and Glucagon (purple, d ). Co-staining between insulin and GPR44 shows up in yellow in the composite image ( e )

Article Snippet: Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa-647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: Confocal Microscopy, Staining

Journal: EJNMMI research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass.

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: Fig. 4 In vitro autoradiography of [11C]AZ12204657 reveals focal binding patterns on pancreatic sections from non-diabetic human donors (a). The binding could be competed away with an excess of GPR44 antagonist AZD3825 (b). Similar GPR44-mediated binding was seen in pancreatic sections from T2D donors (c, d). The focal binding corresponded to Islets of Langerhans as assessed by immunofluorescent insulin staining (e, f) (representative results from three independent experiments on human sections with measurements performed in duplicates). The islet specific targeting was further assessed by [11C]AZ12204657 binding to homogenates of purified human islets of Langerhans and exocrine tissue preparations (results are from two independent experiments on homogenates with measurements performed in triplicates) (g). Binding of [11C]AZ12204657 in pancreatic sections from NHP was similarly focal in nature (h), consistent with the heterogeneous distribution of islets of Langerhans (i), and GPR44-mediated (j) (representative results from six independent experiments on NHP sections with measurements performed in singlets or duplicates)

Article Snippet: Confocal microscopy of GPR44, insulin, and glucagon in human islets of Langerhans Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: In Vitro, Autoradiography, Binding Assay, Staining, Purification

Journal: EJNMMI research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass.

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: Fig. 3 In vitro ligand binding to membranes of HEK293 cells transfected with human recombinant GPR44 and ligand potency at human GPR44 using a clonal beta cell line (EndoC-βH1). [3H]ProstaglandinD2 (a) and [6-3H-phenoxy]-AZ12204657 (b) binding to human GPR44 was displaced with increasing concentrations of AZ12204657. Results are from two independent experiments with three measurements at each concentration and presented as mean ± SD. The potency of AZ12204657 to inhibit the signal of 15(R)-15-methyl-PGD2 in human beta cells measured by the label free DMR assay (c). Results are from two independent experiments with two measurements at each concentration and presented as mean ± SD

Article Snippet: Confocal microscopy of GPR44, insulin, and glucagon in human islets of Langerhans Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: In Vitro, Ligand Binding Assay, Transfection, Recombinant, Binding Assay, Concentration Assay

Journal: EJNMMI research

Article Title: The development of a GPR44 targeting radioligand [ 11 C]AZ12204657 for in vivo assessment of beta cell mass.

doi: 10.1186/s13550-018-0465-6

Figure Lengend Snippet: Fig. 5 Confocal microscopy of a human islet, showing antibody staining for cell nucleus (blue, a), insulin (green, b), GPR44 (red, c) and Glucagon (purple, d). Co-staining between insulin and GPR44 shows up in yellow in the composite image (e)

Article Snippet: Confocal microscopy of GPR44, insulin, and glucagon in human islets of Langerhans Briefly, human islets were stained with a polyclonal rabbit antibody against GPR44 (HPA014259, dilution 1:10, Atlas Antibodies, Stockholm, Sweden), as well as with rat anti-human insulin (MAB1417, dilution 1:100, R&D Systems, Minneapolis, MN, USA) and Alexa647-conjugated mouse anti-glucagon (G2654, dilution 1:500, Sigma-Aldrich).

Techniques: Confocal Microscopy, Staining