Western Blot Analysis of Intestinal Proteins

Isolated small intestinal crypt cells were homogenized in the lysis buffer as previously described (39 (link)). Protein lysates were cleared by spinning the samples twice at 4°C. Subsequently, samples were separated on SDS-PAGE and analyzed by western blotting as previously described (39 (link)). Primary antibodies used are mouse anti-PTBP1 (Life Technologies, 324800), mouse anti-PTBP2 antibody (ABCAM, sc-376316), rabbit anti-Pan AKT (Cell Signaling, 4685), rabbit anti-Phospho-AKT (Ser473) (Cell Signaling, 4060), mouse anti-HSC70 (Santa Cruz Biotechnology, SC-7298), and rabbit anti-GRP78 (Abcam, ab32618). Membranes were incubated with HRP-linked secondary antibodies and developed using ECL prime (G&E Healthcare Life Sciences).

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Chembazhi U.V., Tung W.S., Hwang H., Wang Y., Lalwani A., Nguyen K.L., Bangru S., Yee D., Chin K., Yang J., Kalsotra A, & Mei W. (2023). PTBP1 controls intestinal epithelial regeneration through post-transcriptional regulation of gene expression. Nucleic Acids Research, 51(5), 2397-2414.

Publication 2023

mouse anti-PTBP1 rabbit anti-Pan AKT rabbit anti-Phospho-AKT (Ser473) mouse anti-HSC70 rabbit anti-GRP78 ECL prime

Anti antibody Antibodies Buffer Cells Crypt Grp78 Membranes Mouse Protein Rabbit Sds page Small intestinal

Corresponding Organization : University of Illinois Urbana-Champaign

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Levels of PTBP1 protein
Levels of PTBP2 protein
Levels of Pan AKT protein
Levels of Phospho-AKT (Ser473) protein
Levels of HSC70 protein
Levels of GRP78 protein

control variables

None explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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