Antibody Characterization for Cell Biology

The antibodies used for this study were AMYLASE (A8273; Sigma-Aldrich), Atg5 (PM050, MBL, Woburn, MA), phos-TFEB (ABE1971; Millipore, Burlington, MA), TFEB (A303-673A; Bethyl Laboratories, Montgomery, TX), CK19 (TROMA-III, Developmental Studies Hybridoma Bank, Iowa City, IA), GAPDH (2118; Cell Signaling Technology, Danvers, MA), p62 (H00008878-M01; Abnova, Taipei, Taiwan), phos-4EBP1 (9451; Cell Signaling Technology), 4EBP1 (9452; Cell Signaling Technology), phos-MAPK (9101; Cell Signaling Technology), MAPK (9102; Cell Signaling Technology), phos-S6-ribosomal protein (4858; Cell Signaling Technology), S6-ribosomal protein (2217; Cell Signaling Technology), GFP (sc-9996; Santa Cruz Biotechnology, Dallas, TX), LAMIN A/C (2032; Cell Signaling Technology), LAMP1 (1D4B and H4A3; Developmental Studies Hybridoma Bank, Iowa City, IA), LAMP2 (ABL-93 and H4A4; Developmental Studies Hybridoma Bank), and PGC1α (PAB12061; Abnova). The TAP antibody was generated by Thomas Kolodecik from Yale University.⁴⁵ (link) Antibodies for VATP6V1a and VATP6V1b2 are gifts from Dr. Dennis Brown from Harvard Medical School. The Anti-LC3 antibody was generated as previously described.⁴³ (link) HRP-conjugated, FITC-conjugated and Rhodamine-conjugated secondary antibodies were from Jackson ImmunoResearch (West Grove, PA).

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Wang S., Ni H.M., Chao X., Ma X., Kolodecik T., De Lisle R., Ballabio A., Pacher P, & Ding W.X. (2020). Critical Role of TFEB-Mediated Lysosomal Biogenesis in Alcohol-Induced Pancreatitis in Mice and Humans. Cellular and Molecular Gastroenterology and Hepatology, 10(1), 59-81.

Publication 2020

GAPDH phos-4EBP1 4EBP1 phos-MAPK phos-S6-ribosomal protein S6-ribosomal protein LAMIN A/C PGC1α Rhodamine-conjugated secondary antibodies

4ebp1 Amylase Anti antibody Antibodies Antibody Ck19 Fitc Gapdh Gifts Hybridoma Lamin a c Lamp1 Lamp2 Pgc1α Rhodamine Ribosomal protein Tfeb

Corresponding Organization : University of Kansas Medical Center

Other organizations : Yale University, VA Connecticut Research and Education Foundation, Baylor College of Medicine, Telethon Institute Of Genetics And Medicine, Neurological Research Institute, Texas Children's Hospital, Federico II University Hospital, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

None explicitly mentioned

control variables

The antibodies used for this study: AMYLASE, Atg5, phos-TFEB, TFEB, CK19, GAPDH, p62, phos-4EBP1, 4EBP1, phos-MAPK, MAPK, phos-S6-ribosomal protein, S6-ribosomal protein, GFP, LAMIN A/C, LAMP1, LAMP2, PGC1α, TAP, VATP6V1a, VATP6V1b2, and LC3

controls

Positive control: None mentioned
Negative control: None mentioned

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