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and iii) a mono or polyglutamate chain of variable length (Fig

4D and E

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The membrane was then incubated with the Horseradish peroxidase-conjugated anti-rabbit IgG secondary antibody (1:5000, in 5%milk in TBST) or anti-mouse IgG secondary (1:5000, in 5%milk in TBST) at room temperature for 1h, followed with washing and detection using the enhanced chemiluminescence (ECL) detection kit (Cytiva)