Western Blot Analysis of Pancreatic and Hepatic Proteins

In the pancreas and liver tissues of 2-, 4- and 7-day-old neonates, western blot analysis of insulin promoter factor, duodenal homeobox gene-1 (PDX-1) and anti-apoptosis factor B-cell lymphoma-2 (Bcl-2) was carried out as previously described (18 (link)). Primary antibodies included rabbit anti-PDX-1 diluted at 1:4,000 (#2437, Cell Signaling Technology), rabbit anti-Bcl-2 diluted at 1:4,000 (AB1722, Millipore), rabbit anti-betatrophin diluted at 1:4,000 (ab180915, Abcam), rabbit anti-Bax diluted at 1:1,000 (#2772T, Cell Signaling Technology) and rabbit anti-β-tubulin diluted at 1:2,000 (#2128, Cell Signaling Technology). The intensity of bands was analyzed using Quantity One Software (Bio-Rad). Peroxidase-conjugated secondary antibody was diluted at 1:8,000 (ZB2301, ZSGB-BIO, Beijing, China). The protein concentration was measured using Quantity One Software (Bio-Rad).

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Zhang D., Yu Y.J., Xu F.S., Yuan J.H., Wang R., Zhang C.S., Wang L.X., Liu Y., Song L.M., Liu J.L, & Dong J. (2019). Recombinant betatrophin (Angptl-8/lipasin) ameliorates streptozotocin-induced hyperglycemia and β-cell destruction in neonatal rats. Molecular Medicine Reports, 20(5), 4523-4532.

Publication 2019

ab180915 Quantity One Software ZB2301

Antibodies Antibody Apoptosis B cell lymphoma Betatrophin Duodenal Homeobox gene Insulin Liver Neonates Pancreas Pdx 1 Peroxidase Protein Rabbit Tissues Tubulin Western blot analysis

Corresponding Organization :

Other organizations : Qingdao University, Jiaozuo University, Qingdao Municipal Hospital, Affiliated Hospital of Qingdao University, McGill University Health Centre

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Variable analysis

independent variables

Age of neonates (2 days, 4 days, 7 days)

dependent variables

Protein expression levels of PDX-1
Protein expression levels of Bcl-2
Protein expression levels of betatrophin
Protein expression levels of Bax

control variables

Tissue type (pancreas and liver)
Primary antibody dilutions
Secondary antibody dilution
Protein concentration measurement method

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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