[IF=5.279] Tao Wu. et al. Lactobacillus rhamnosus LRa05 Ameliorate Hyperglycemia through a Regulating Glucagon-Mediated Signaling Pathway and Gut Microbiota in Type 2 Diabetic Mice. J Agr Food Chem. 2021;XXXX(XXX):XXX-XXX  WB ;  Mouse.  

PubMed:34340304

[IF=3.73] Rafacho, Alex, et al. "Pancreatic Alpha-Cell Dysfunction Contributes to the Disruption of Glucose Homeostasis and Compensatory Insulin Hypersecretion in Glucocorticoid-Treated Rats." PLOS ONE 9.4 (2014): e93531.  IHC-P ;  Rat.  

PubMed:24705399

Sample: Cerebellum (Mouse) Lysate at 40 ug
Primary: Anti-Glucagon Receptor (bs-3945R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD

Sample:Kidney (Mouse) Lysate at 40 ug
Primary: Anti-Glucagon Receptor(bs-3945R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51kD
Observed band size: 51kD

Paraformaldehyde-fixed, paraffin embedded (human stomach tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Glucagon Receptor) Polyclonal Antibody, Unconjugated (bs-3945R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Glucagon Receptor) Polyclonal Antibody, Unconjugated (bs-3945R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control:HepG2.
Primary Antibody (green line): Rabbit Anti-Glucagon Receptor antibody (bs-3945R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Protocol
The cells were incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature.Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control(blue): Molt-4 Cells(fixed with 2% paraformaldehyde (10 min)).
Primary Antibody: Rabbit Anti-Glucagon Receptor/AF647 Conjugated antibody (bs-3945R/AF647), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/AF647(orange) ,used under the same conditions.