Anti-GIP Antibody Picoband®

Western blot analysis of GIP using anti-GIP antibody (PB9946).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human HepG2 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GIP antigen affinity purified polyclonal antibody (Catalog # PB9946) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GIP at approximately 17 kDa. The expected band size for GIP is at 17 kDa.

IHC analysis of GIP using anti-GIP antibody (PB9946).
GIP was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-GIP Antibody (PB9946) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of GIP using anti-GIP antibody (PB9946).
GIP was detected in a paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-GIP Antibody (PB9946) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Alterations of the protein levels of the components of the GSK-3β—β-catenin—TCF7L2—GLP-1 axis under varying glucose conditions. ( A ) Molecular structure of Emodin. ( B ) Relative protein levels of p-GSK-3β, GSK-3β, β-catenin and TCF7L2 in Control and Emodin groups under different glucose conditions, detected by the western blot analysis. ( C ) Relative mRNA expression of β-catenin and TCF7L2 in Control and Emodin groups under different glucose conditions, detected by real-time PCR. ( D ) GLP-1 and GIP concentration measured by ELISA in the cell supernatant in Control and Emodin groups under different glucose conditions. *p < 0.05, **p < 0.01 vs control under no glucose, # p < 0.05, ## p < 0.01 vs control under low glucose, &p < 0.05, && p < 0.01 vs control under high glucose; mean ± SEM. ( E,F ) Inhibition of TCF7L2 and GSK-3β abolished DMC’s effects. ( E ) Effects of emodin (the main component of DMC) and SB216763 (a specific GSK-3β inhibitor) on the protein levels of β-catenin and TCF7L2 under HG (high glucose) conditions. ( F ) Relative mRNA expression of β-catenin and TCF7L2 in the three groups. ( G ) Relative protein expression of TCF7L2 in HG, HG + emodin and HG + emodin + TCF7L2 SiRNA groups. ( H ) GLP-1 and GIP concentration measured by ELISA in the cell supernatant in the three groups. *p < 0.05, **p < 0.01 vs HG, #p < 0.05, ##p < 0.01 vs HG + Emodin; mean ± SEM.
Index in PubMed under a CC BY license. PMID: 27721485

GLP-1 and GIP concentrations in the serum and mRNA levels of GCG and GIP. ( A ) GLP-1 concentrations in the serum in the in the Control, DM and DMC groups. ( B ) GCG (the gene encoding GLP-1) mRNA levels in ileum. ( C ) GIP concentrations in the serum in the five groups. ( D ) GIP mRNA levels in ileum. ( E ) Insulin levels in the serum in the five groups. *p < 0.05, **p < 0.01 vs Control, # p < 0.05, ## p < 0.01 vs DM; mean ± SEM.
Index in PubMed under a CC BY license. PMID: 27721485

Expression of GLP-1 and GIP in ileum. ( A ) Images (×400) of immunofluorescence staining of GLP-1 (glucagon-like peptide-1) and GIP (gastric inhibitory polypeptide) expression in ileum in the Control, DM and DMC groups. ( B ) Immunofluorescence results (×400) indicating GIP expression. (scale bar: 20μm). ( C ) Quantification of the GLP-1 fluorescence intensity (integrated density per stained area). ( D ) Quantification of the GIP fluorescence intensity (integrated density per stained area). *p < 0.05 vs Control, # p < 0.05 vs DM; mean ± SEM.
Index in PubMed under a CC BY license. PMID: 27721485