Quantitative Protein Analysis with Infrared Scanner

Various proteins, include crude homogenate of cortex and isolated glomeruli were quantitated (Epoch Gen5, BioTek, Winooski, VT), and mixed at 1:1 with SDS sample buffer following our published method [20 (link)]. The membrane was incubated with IRDye®800CW labeled secondary antibody for target protein and IRDye® 680LT anti-mouse IgG antibody (LI-COR, Lincoln, NE). The membrane was simultaneously scanned at both wave lengths on an infrared fluorescence scanner (Odyssey, LI-COR), with target protein as green and control α-actin as red. Each band was digitally quantitated as IOD.

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Wu J., Carlock C., Shim J., Moreno-Gonzalez I., Glass W I.I., Ross A., Barichello T., Quevedo J, & Lou Y. (2021). Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau. Molecular Psychiatry, 26(10), 5912-5924.

Publication 2021

Epoch Gen5 IRDye® 680LT anti-mouse IgG antibody infrared fluorescence scanner

Actin Anti antibody Antibody Buffer Cortex Epoch Fluorescence Glomeruli Irdye 800cw Membrane Mouse Protein target Proteins

Corresponding Organization :

Other organizations : The University of Texas Health Science Center at Houston

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Variable analysis

independent variables

Various proteins, including crude homogenate of cortex and isolated glomeruli

dependent variables

Protein quantitation

control variables

SDS sample buffer
IRDye®800CW labeled secondary antibody for target protein
IRDye® 680LT anti-mouse IgG antibody
Infrared fluorescence scanner (Odyssey, LI-COR)

controls

Control α-actin as red

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