Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Primary Antibodies and Staining Settings in the Verification of Immunohistochemical Somatostatin Receptor 1–5 Staining Protocols.

Article Snippet: SSTR1 , AbD Serotec (MCA5924) , sstr1 , Western blot/transfected HEK-293 cell line 11 , Mouse , Tris-EDTA pH 9.0 , 1:100 , 30 min , Envision , LabVision , clone sstr1 Citrate pH 6.0 1:500 30 min/RT Envision.

Techniques: Staining, Immunohistochemical staining, Clone Assay, Incubation, Western Blot, Affinity Purification, Transfection

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Somatostatin receptor 1–5 expression in pancreatic (A, C, E, G, I) and small intestine (B, D, F, H, J) neuroendocrine cells. Scale bar: 50 µm; magnification: 40×.

Article Snippet: SSTR1 , AbD Serotec (MCA5924) , sstr1 , Western blot/transfected HEK-293 cell line 11 , Mouse , Tris-EDTA pH 9.0 , 1:100 , 30 min , Envision , LabVision , clone sstr1 Citrate pH 6.0 1:500 30 min/RT Envision.

Techniques: Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Somatostatin Receptor 1–5 Expression in Neuroendocrine Tumors.

Article Snippet: SSTR1 , AbD Serotec (MCA5924) , sstr1 , Western blot/transfected HEK-293 cell line 11 , Mouse , Tris-EDTA pH 9.0 , 1:100 , 30 min , Envision , LabVision , clone sstr1 Citrate pH 6.0 1:500 30 min/RT Envision.

Techniques: Expressing, Membrane, Biomarker Discovery

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Primary Antibodies and Staining Settings in the Verification of Immunohistochemical Somatostatin Receptor 1–5 Staining Protocols.

Article Snippet: Rabbit Tris-EDTA pH 9.0 1:50 1:100 1:500 30 min Envision Labvision Citrate pH 6.0 1:50 1:100 1:500 30 min Envision Labvision Abcam (ab137026) UMB5 Western blot/ sstr3-transfected HEK-293 cell line 5 Rabbit Citrate pH 6.0 1:7000 60 min Envision Labvision SSTR4 AbD Serotec (MCA5922) sstr4 Western blot/ transfected HEK-293 cell line 11 Mouse Tris-EDTA pH 9.0 1:500 30 min Envision Labvision clone sstr4 Citrate pH 6.0 1:500 30 min/RT Envision Citrate pH 6.0 1:500 30 min Envision Labvision Standard CC1 1:250 1:500 32 min UltraVIEW DAB BenchMark XT CC1 64 min 1:100 1:500 32 min OptiVIEW DAB BenchMark XT SSTR5 Bio-RAD (MCA5923) sstr5 Western blot/ transfected HEK-293 cell line 11 Mouse Tris-EDTA pH 9.0 1:100 30 min Envision Labvision clone sstr5 Citrate pH 6.0 1:1000 30 min/RT Envision Citrate pH 6.0 1:50 1:100 1:500 1:1000 30 min Envision Labvision CC1 64 min 1:100 1:500 32 min OptiVIEW DAB BenchMark XT Abcam (ab109495) UMB4 Western blot/ sstr5-trasfected HEK-293 cell line 4 Rabbit Citrate pH 6.0 1:100 1:500 1:1000 30 min Envision Labvision clone UMB4 Citrate pH 6.0 1:1000 30 min/RT Envision Tris-EDTA pH 9.0 1:100 1:500 1:1000 30 min Envision Labvision Open in a separate window Abbreviations: SSTR, somatostatin receptors.

Techniques: Staining, Immunohistochemical staining, Clone Assay, Incubation, Western Blot, Affinity Purification, Transfection

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Somatostatin Receptor 1–5 Expression in Neuroendocrine Tumors.

Article Snippet: Rabbit Tris-EDTA pH 9.0 1:50 1:100 1:500 30 min Envision Labvision Citrate pH 6.0 1:50 1:100 1:500 30 min Envision Labvision Abcam (ab137026) UMB5 Western blot/ sstr3-transfected HEK-293 cell line 5 Rabbit Citrate pH 6.0 1:7000 60 min Envision Labvision SSTR4 AbD Serotec (MCA5922) sstr4 Western blot/ transfected HEK-293 cell line 11 Mouse Tris-EDTA pH 9.0 1:500 30 min Envision Labvision clone sstr4 Citrate pH 6.0 1:500 30 min/RT Envision Citrate pH 6.0 1:500 30 min Envision Labvision Standard CC1 1:250 1:500 32 min UltraVIEW DAB BenchMark XT CC1 64 min 1:100 1:500 32 min OptiVIEW DAB BenchMark XT SSTR5 Bio-RAD (MCA5923) sstr5 Western blot/ transfected HEK-293 cell line 11 Mouse Tris-EDTA pH 9.0 1:100 30 min Envision Labvision clone sstr5 Citrate pH 6.0 1:1000 30 min/RT Envision Citrate pH 6.0 1:50 1:100 1:500 1:1000 30 min Envision Labvision CC1 64 min 1:100 1:500 32 min OptiVIEW DAB BenchMark XT Abcam (ab109495) UMB4 Western blot/ sstr5-trasfected HEK-293 cell line 4 Rabbit Citrate pH 6.0 1:100 1:500 1:1000 30 min Envision Labvision clone UMB4 Citrate pH 6.0 1:1000 30 min/RT Envision Tris-EDTA pH 9.0 1:100 1:500 1:1000 30 min Envision Labvision Open in a separate window Abbreviations: SSTR, somatostatin receptors.

Techniques: Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Primary Antibodies and Staining Settings in the Verification of Immunohistochemical Somatostatin Receptor 1–5 Staining Protocols.

Article Snippet: SSTR4 , AbD Serotec (MCA5922) , sstr4 , Western blot/ transfected HEK-293 cell line 11 , Mouse , Tris-EDTA pH 9.0 , 1:500 , 30 min , Envision , Labvision , , clone sstr4 Citrate pH 6.0 1:500 30 min/RT Envision.

Techniques: Staining, Immunohistochemical staining, Clone Assay, Incubation, Western Blot, Affinity Purification, Transfection

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Immunohistochemical Expression of Somatostatin Receptor Subtypes in a Panel of Neuroendocrine Neoplasias

doi: 10.1369/0022155419856900

Figure Lengend Snippet: Somatostatin Receptor 1–5 Expression in Neuroendocrine Tumors.

Article Snippet: SSTR4 , AbD Serotec (MCA5922) , sstr4 , Western blot/ transfected HEK-293 cell line 11 , Mouse , Tris-EDTA pH 9.0 , 1:500 , 30 min , Envision , Labvision , , clone sstr4 Citrate pH 6.0 1:500 30 min/RT Envision.

Techniques: Expressing, Membrane, Biomarker Discovery

Journal: International Journal of Molecular Sciences

Article Title: FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes

doi: 10.3390/ijms18112234

Figure Lengend Snippet: Cytofluorimetric analysis of islet-like aggregates. PANC-1 cells were treated with 50% trypsin for 30 s and incubated with serum-free medium supplemented with 0.1% BSA (0.1%) plus 1.1 mg/L transferrin (SDT). Analysis was expressed as percentage of positive cells in association with the specific markers. SDT medium was then supplemented with 500 ng/mL FGF-2b or 500 ng/mL hPL-A, or both hormones (FGF-2b plus hPL-A). After 96 h, islet-like aggregates were disaggregated to form single cell suspensions. Then, cells were fixed, permeabilized and stained for insulin, PDX-1, Nkx2.2, Nkx6.1, somatostatin, glucagon, MUC-1, Cytokeratin-19, and Glut-2, and immediately acquired on a BD FACSCalibur (at least 5 × 10 4 event). # p < 0.05 vs. SDT; n = 4 (four separate experiments). Control: untreated PANC-1 cells.

Article Snippet: Inc., Heidelberg, Germany); and mouse anti-human MUC-1, mouse anti-human somatostatin (Ab-Serotec, Oxford, UK).

Techniques: Incubation, Staining, Control

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Cplx 2 is expressed in mouse pancreatic islet β-, α- and δ-cells, and loss of Cplx 2 protein expression is observed in Cplx 2 KO mouse islets. (A) Representative images of RNA fluorescence in- situ hybridization (FISH) of Cplx 2 (green puncta) transcripts in fixed WT mouse islets co-immunostained with insulin (INS) for β-cells (upper panel), glucagon (GCG) for α-cells (middle panel), and somatostatin (SST) for δ-cells (bottom panel). (B) Comparison of Cplx 2 protein expression in fixed WT and Cplx 2 KO mouse pancreatic islet sections immunostained with antibodies against Cplx 2 (cyan) and either insulin (INS), glucagon (GCG) or somatostatin (SST) (magenta). Scale bars are 20 µm.

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques: Expressing, Fluorescence, In Situ Hybridization, Comparison

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Effect of loss of Cplx 2 on insulin, glucagon and somatostatin secretion from intact mouse islets and dispersed islet cells. Insulin (A), glucagon (B) and somatostatin (C) secretions were measured in wild-type (WT) and Cplx 2 KO (KO) intact mouse islets at low (1 mM, white) and high (11 mM, gray) glucose concentrations. Insulin (D), glucagon (E) and somatostatin (F) secretions were measured in WT and KO dispersed cells at low (1 mM, white) and high (11 mM, gray) glucose concentrations. Data presented as mean ± SD (n > 2 experiments). Statistical significance between different glucose concentrations either in WT or KO islets was determined using unpaired t test ***P < 0.001; **P < 0.01; *P < 0.05. Statistical significance between WT and KO at the same glucose concentration was measured with unpaired t test ##P < 0.01; #P < 0.05.

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Loss of Cplx 2 leads to deficiency in paracrine inhibition of insulin secretion by somatostatin. Insulin (A) and glucagon (B) secretion from WT and Cplx 2 KO (KO) intact mouse islets at low (1 mM, white) and high (11 mM, gray) glucose concentrations with or without 1 µM somatostatin (SST) treatment. Insulin (C) and glucagon (D) secretion from WT and KO dispersed cells at low (1 mM, white) and high (11 mM, gray) glucose concentrations with or without 1 µM SST treatment. Data presented as mean ± SD (n > 2 experiments). Statistical significance between conditions was determined using unpaired t test ***P < 0.001; **P < 0.01; *P < 0.05.

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques: Inhibition

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Effects of antagonism of insulin receptor (IR) and type 2 somatostatin receptor (SSTR2) are similar for insulin and glucagon secretion from Cplx 2 KO intact and dispersed mouse islets. Insulin (A) and glucagon (B) secretion from WT and Cplx 2 KO (KO) intact mouse islets at low (1 mM, white) and high (11 mM, gray) glucose concentrations in the presence or absence of 2 µM S961 (IR antagonist) or 100 nM CYN 154806 (CYN, SSTR2 antagonist). Insulin (C) and glucagon (D) secretion from WT and KO dispersed cells at low (1 mM, white) and high (11 mM, gray) glucose concentrations in the presence or absence of 2 µM S961 or 100 nM CYN. Data presented as mean ± SD (n > 2 experiments). Statistical significance between conditions was determined using unpaired t test ***P < 0.001; **P < 0.01; *P < 0.05.

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques:

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Cplx 2 mediates paracrine regulation of β-cell Ca 2+ activity by somatostatin. Representative intensity traces of Fluo-4 Ca 2+ indicator dye in β-cells of WT (A) and KO (B) intact mouse islets with the respective treatment conditions indicated by dashed lines. (C) Percentage change in Fluo-4 intensity averaged over traces acquired at 11 mM glucose concentration in the presence and absence of 1 μM SST for β-cells of WT and KO islets (n > 2 islets, 29 cells). Representative intensity traces of GCaMP6f fluorescence in α-cells of WT (D) and KO (E) islets with treatment conditions indicated by dashed lines. (F) Treatment with 1 μM SST inhibits α-cell Ca 2+ activity in both WT and KO islets under low (1 mM, white) and high (11 mM, gray) glucose levels (n > 2 islets, 31 cells). Data presented as mean ± SD. Statistical significance between conditions was determined using unpaired t test ***P < 0.001; **P < 0.01; *P < 0.05.

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques: Activity Assay, Concentration Assay, Fluorescence

Journal: bioRxiv

Article Title: Role of Complexin 2 in the regulation of hormone secretion from the islet of Langerhans

doi: 10.1101/2024.10.28.620710

Figure Lengend Snippet: Loss of Cplx 2 dysregulates paracrine inhibition of insulin vesicle fusion by somatostatin. (A) A representative TIRF time trace of an isolated granule fusion event in a VAMP2-superecliptic pHluorin (SEP) adenovirus transduced Cplx 2 KO islet. Scale bar is 1 μm. (B) Insulin granule fusion events in WT and KO VAMP2-SEP islets at low (1 mM) and high glucose (11 mM) concentrations with and without 1 μM SST (n > 2 islets, 11 cells). (C) Half-life (t 1/2 ) of fusion events from (B) for WT and KO islets (n > 21 fusion events).

Article Snippet: Pancreas sections were then incubated with primary antibodies diluted in blocking buffer against either insulin (2D11-H5; 1:100; Santa Cruz), glucagon (MAB1249; 1:100; R&D Systems) or somatostatin (MAB2358; 1:50; R&D Systems) along with Cplx 2 (18149-1-AP; 1:50; Proteintech) overnight at 4°C.

Techniques: Inhibition, Isolation