This is a knockout-validated antibody summary, based on the publications "Functional CRISPR screening identifies the ufmylation pathway as a regulator of SQSTM1/p62", as cited below [1], "Cdk5rap3 is essential for intestinal Paneth cell development and maintenance" for western blot knockout validation (figure 5f) [2], and "Cdk5rap3 is essential for intestinal Paneth cell development and maintenance" for western blot knockout validation (figure 5f) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot of UFM1 in control and UFM1-KO H4 neuroglioma cell line (stably expressing Cas9 and GFP-SQSTM1). GAPDH was used as loading control. From [1].

Rabbit monoclonal IgG

Company: Abcam

Antibody: UFM1

Catalog number: ab109305

Summary: Rabbit monoclonal IgG against a synthetic peptide, corresponding to residues from C terminal of human UFM1. Reacts with human, mouse and rat. Suitable for western blot, immunoprecipitation and immunohistochemistry (paraffin). Unsuitable for immunocytochemistry and flow cytometry.

Western blot

Control and UFM1-KO H4 neuroglioma cell line (stably expressing Cas9 and GFP-SQSTM1). Cells were lysed in RIPA buffer (Thermo Scientific, Waltham, MA) supplemented with protease inhibitor cocktail (Roche) followed by sonication. Alternatively, protein lysates were prepared in RIPA buffer containing protease inhibitors and 1% SDS followed by purification with QIAshredder columns (Qiagen).

Odyssey blocking buffer for 1 hr or PBS/0.1% Tween-20/5% milk.

Diluted in Odyssey blocking buffer supplemented with 0.1% Tween-20 (Bio-Rad) or in PBS/0.1% Tween-20/5% milk overnight at 4°C.

Anti-rabbit IRDye 800CW (LI-COR) diluted in Odyssey blocking buffer supplemented with 0.1% Tween-20 or anti-rabbit HRP (Thermo Fisher Scientific) diluted in PBS/0.1% Tween-20/5% milk incubated for 1 hr at room temperature.

Odyssey infrared imaging system or SuperSignal West Pico chemiluminescent substrate (Thermo Scientific) and autoradiography.