Western Blot Analysis of Pancreatic Tumor

Tumor lysates were made using normal human pancreas tissue and the pancreatic cancer cell line BxPC-3-GFP xenograft. Western blotting was performed as previously described.^{18 (link)} Protein samples were isolated and transferred to Trans-Blot Turbo mini nitrocellulose membranes (Bio-Rad, Hercules, CA, Cat. 1704270) using the Bio-Rad Trans-Blot Turbo transfer system. The membrane was then incubated with the IRDye700DX-conjugated antibody at 4 °C overnight. The membrane was scanned with an Odyssey infrared imaging system (model 9120; LI-COR), and detection and quantification of band intensities were conducted using Image Studio Lite software (version 5.2; LI-COR). β-Actin was used as standard.

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Nishino H., Turner M.A., Amirfakhri S., Lwin T.M., Hosseini M., Singer B.B., Hoffman R.M, & Bouvet M. (2022). Proof of Principle of Combining Fluorescence-Guided Surgery with Photoimmunotherapy to Improve the Outcome of Pancreatic Cancer Therapy in an Orthotopic Mouse Model. Annals of Surgical Oncology, 30(1), 618-625.

Publication 2022

Trans-Blot Turbo mini nitrocellulose membranes Trans-Blot Turbo transfer system Odyssey infrared imaging system Image Studio Lite software

Actin Antibody Cell line Human Link protein Membrane Nitrocellulose Normal tissue Pancreas Pancreatic cancer Tumor Xenograft

Corresponding Organization : University of California, San Diego

Other organizations : Kyoto University, Dana-Farber Cancer Institute, Boston University, University of Duisburg-Essen

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Variable analysis

independent variables

Normal human pancreas tissue
Pancreatic cancer cell line BxPC-3-GFP xenograft

dependent variables

Protein expression levels (quantified by band intensities)

control variables

Experimental procedures for Western blotting as previously described
Use of Trans-Blot Turbo mini nitrocellulose membranes
Use of Bio-Rad Trans-Blot Turbo transfer system
Incubation with IRDye700DX-conjugated antibody at 4 °C overnight
Scanning with Odyssey infrared imaging system
Detection and quantification using Image Studio Lite software
Use of β-Actin as standard

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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