Western blot

Published: 2 July 2025| Version 1 | DOI: 10.17632/4g722x4jfg.1
Contributor:
Yulong Wei

Description

Western blot of CRISPR-Cas9-mediated nontargeting (NT) versus BACH2 knockout 3 days after CRISPR-Cas9-gRNA nucleofection of human gut cells.

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Steps to reproduce

Protein lysates were extracted using RIPA lysis/protein extraction buffer with complete mini protease inhibitor cocktail (Roche). Lysates were qualified using Pierce BCA Assay (ThermoFisher). Western blot was performed using BACH2 (D3R3G) Rabbit mAb (1:1000 dilution), β-Actin (D6A8) Rabbit mAb (1:1000 dilution), and anti-rabbit IgG, HRP-linked antibody (1:1000 dilution), developed with Amersham ECL Select Western Blotting Detection Reagent (Cytiva). The blot was visualized by exposure to X-ray films (Fuji Film) for 30s.

Institutions

  • Yale University

Categories

Gut, Western Blot, T Cell, CRISPR/Cas9

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