mouse monoclonal primary antisera  (OriGene)

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 2. Effects of intra-VMN administration of SCR versus GPbb or GPmm siRNA on VMN GPbb and GPmm protein expression. VMN tissue was bilaterally micropunch-dissected from rostral (left-hand column), middle (middle column), and caudal (right-hand column) VMN fresh-frozen tissue sections after sc V or INS injection into SCR-, GPbb, or GPmm-siRNA-pretreated animals. Data show mean normalized GPbb (A, C, E) or GPmm (B, D, F) protein OD measures ± SEM for the following treatment groups (n = 4 per group): SCR/V (solid white bars), GPbb siRNA/V (diagonal-striped white bars), GPmm siRNA/V (cross-hatched white bars), SCR/INS (solid gray bars), GPbb siRNA/INS (diagonal-striped gray bars), and GPmm siRNA/INS (cross-hatched gray bars). *p < .05; **p < .01; ***p < .001. Abbreviations: VMN = ventromedial hypothalamic nucleus; SCR = scramble; GP = glycogen phosphorylase; GPbb = glycogen phosphorylase- brain type; GPmm = glycogen phosphorylase-muscle type; OD = optical density; SEM = standard error of mean; INS = insulin; V = vehicle; sc = subcutaneous.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Expressing, Injection

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 3. Effects of GP isoform knockdown on VMN NO nerve cell transmitter marker expression. For each animal, nNOS-immunopositive neurons were collected by laser-catapult microdissection and pooled separately within rostral (left-hand column), middle (middle column), and caudal (right-hand column) VMN segments. Data depict mean normalized nNOS protein OD measures ± SEM for rostral (A), middle (B), and caudal (C) VMN NO neurons from SCR/V (solid white bars), GPbb siRNA/V (diagonal-striped white bars), GPmm siRNA/V (cross-hatched white bars), SCR/INS (solid gray bars), GPbb siRNA/INS (diagonal-striped gray bars), and GPmm siRNA/INS (cross-hatched gray bars) treatment groups. *p < .05; **p < .01; ***p < .001. Abbreviations: GP = glycogen phosphorylase; VMN = ventromedial hypothalamic nucleus; NO = nitric oxide; nNOS = neuronal nitric oxide synthase; OD = optical density; SEM = standard error of measurement; INS = insulin; SCR = scramble.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Knockdown, Marker, Expressing, Laser Capture Microdissection

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 4. GPbb versus GPmm regulation of VMN NO neuron total and phosphorylated AMPK protein expression. Western blot analysis of rostral (left-hand column), middle (middle column), and caudal (right-hand column) VMN NO neurons was performed to determine effects of GP isoform knockdown on total AMPK and pAMPK protein expression during glucostasis versus glucoprivation. For each VMN segment, data show mean normalized AMPK (A, C, E) or pAMPK (B, D, F) protein OD measures ± SEM for NO neurons collected from SCR-, GPbb-, or GPmm siRNA-pretreated animals after sc injection of V or INS. *p < .05; **p < .01; ***p < .001. Abbreviations: GPbb = glycogen phosphorylase-brain type; GPmm = glycogen phosphorylase-muscle type; VMN = ventromedial hypothalamic nucleus; NO = nitric oxide; AMPK = 5′-AMP-activated protein kinase; OD = optical density; SEM = standard error of mean; pAMPK = phosphorylated AMPK; SCR = scramble; INS = insulin; V = vehicle; sc = subcutaneous.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Expressing, Western Blot, Knockdown, Injection

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 5. Effects of GP isoform knockdown on VMN GABA neuron GAD protein expression. Data depict mean normalized GAD protein OD values ± SEM for GAD-immunolabeled neurons harvested from rostral (A), middle (B), and caudal (C) VMN segments of SCR/V (solid white bars), GPbb siRNA/V (diagonal-striped white bars), GPmm siRNA/V (cross-hatched white bars), SCR/INS (solid gray bars), GPbb siRNA/INS (diagonal-striped gray bars), and GPmm siRNA/INS (cross-hatched gray bars) treatment groups. *p < .05; **p < .01; ***p < .001. Abbreviations: GP = glycogen phosphorylase; VMN = ventromedial hypothalamic nucleus; GABA = γ-aminobutyric acid; GAD = glutamate decarboxylase65/67; OD = optical density; SEM = standard error of mean; SCR = scramble; INS = insulin; V = vehicle; sc = subcutaneous.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Knockdown, Expressing, Immunolabeling

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 6. GPbb versus GPmm regulation of VMN GABA neuron total and phosphorylated AMPK protein expression. Western blot analysis of rostral (left-hand column), middle (middle column), and caudal (right-hand column) VMN GABA neurons was performed to determine effects of GP isoform knockdown on total AMPK and pAMPK protein expression during glucostasis versus glucoprivation. For each VMN segment, data show mean normalized AMPK (A, C, E) or pAMPK (B, D, F) protein OD measures ± SEM for GAD-immunopositive neurons collected from SCR-, GPbb-, or GPmm siRNA-pretreated animals after sc injection of V or INS. *p < .05; **p < .01; ***p < .001. Abbreviations: GPbb = glycogen phosphorylase-brain type; GPmm = glycogen phosphorylase-muscle type; VMN = ventromedial hypothalamic nucleus; GABA = γ-aminobutyric acid; NO = nitric oxide; AMPK = 5′-AMP-activated protein kinase; OD = optical density; SEM = standard error of mean; pAMPK = phosphorylated AMPK; GAD = glutamate decarboxylase65/67; SCR = scramble; INS = insulin; V = vehicle; sc = subcutaneous.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Expressing, Western Blot, Knockdown, Injection

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 7. Effects of intra-VMN GP siRNA administration on PVN, DMN, and ARH hypothalamic nucleus and LHA GP variant protein expression. Data show mean normalized GPbb (at right) or GPmm (at left) PVN (A), DMN (B), LHA (C), and ARH (D) protein OD values ± SEM after administration of SCR, GPbb siRNA, or GPmm siRNA to the VMN. Abbreviations: VMN = ventromedial hypothalamic nucleus; GP = glycogen phosphorylase; PVN = paraventricular; DMN = dorsomedial; ARH = arcuate; LHA = lateral hypothalamic area; GPbb = glycogen phosphorylase-brain type; GPmm = glycogen phosphorylase-muscle type; OD = optical density; SEM = standard error of mean; SCR = scramble.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Variant Assay, Expressing

Journal: ASN neuro

Article Title: Glycogen Phosphorylase Isoform Regulation of Ventromedial Hypothalamic Nucleus Gluco-Regulatory Neuron 5'-AMP-Activated Protein Kinase and Transmitter Marker Protein Expression.

doi: 10.1177/17590914211035020

Figure Lengend Snippet: Figure 8. VMN GPbb versus GPmm regulation of plasma glucose concentrations and basal and hypoglycemia-associated patterns of counter-regulatory hormone secretion. Data show mean glucose (A), glucagon (B), and corticosterone (C) concentrations ± SEM for the following treatment groups: SCR/V (solid white bars), GPbb siRNA/V (diagonal-striped white bars), GPmm siRNA/V (cross-hatched white bars), SCR/INS (solid gray bars), GPbb siRNA/INS (diagonal-striped gray bars), and GPmm siRNA/INS (cross-hatched gray bars). *p < .05; **p < .01; ***p < .001. Abbreviations: VMN = ventromedial hypothalamic nucleus; GP = glycogen phosphorylase; GPbb = glycogen phosphorylase-brain type; GPmm = glycogen phosphorylase-muscle type; SEM = standard error of mean; SCR = scramble; INS = insulin; V = vehicle; sc = subcutaneous.

Article Snippet: Membranes were blocked with TBS containing 0.1% Tween-20 (Amresco LLC, Solon, OH) and 2.0% bovine serum albumin (MP Biomedicals LLC, Santa Ana, CA), then incubated (36–42 h; 4°C) with rabbit primary polyclonal antisera against GPbb (1:2,000; prod. no. NBP1-32799; Novus Biologicals, Littleton, CO), GPmm (1:2,000; prod. no. NBP2-16689; Novus Biol.), GAD (1:10,000; prod. no. ABN904; Millipore Sigma, Burlington, MA), nNOS (1:1,500; prod. no. NBP1-39681; Novus Biol.), AMPKα1,2 (prod. no. 2532S, 1:2,000; Cell Signaling Technology Inc., Danvers, MA), or phosphoAMPKα1/2 (Thr 172) (pAMPK; prod. no. 2535S, 1:2,000; Cell Signaling Technol.).

Techniques: Clinical Proteomics