Immunoblot Analysis of Cellular Fractions

Immunoblot analysis of total lysate, cytoplasmic and nucleoplasmic fractions was essentially performed as described [29 (link)]. Briefly, primary antibodies were polyclonal antibodies against HISTONE 3 (Agrisera, Vännäs, Sweden, AS10710; rabbit; dilution 1:5000), PHOSPHOENOLPYRUVATE CARBOXYLASE (Agrisera AS09458; rabbit; dilution 1:20,000) and SERRATE (Agrisera AS09532A; rabbit; dilution 1:1000). Secondary antibody was HRP-coupled anti-rabbit IgG (Sigma-Aldrich, Burlington, MA, USA, A0545; dilution 1:2500). Chemiluminescence detection of the immunoblots was done with Fusion-FX6 (Vilber) system.

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Lüders J., Winkel A.R., Reichel M., Bitterer V.W., Scheibe M., Widmann C., Butter F, & Köster T. (2022). Identification of Pri-miRNA Stem-Loop Interacting Proteins in Plants Using a Modified Version of the Csy4 CRISPR Endonuclease. International Journal of Molecular Sciences, 23(16), 8961.

Publication 2022

AS09458 AS09532A HRP-coupled anti-rabbit IgG Fusion-FX6

Anti igg Antibodies Antibody Chemiluminescence Cytoplasmic Dilution Histone Immunoblots Phosphoenolpyruvate carboxylase Rabbit

Corresponding Organization : Bielefeld University

Other organizations : Institute of Molecular Biology

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Variable analysis

independent variables

Primary antibodies used for immunoblot analysis

dependent variables

Chemiluminescence detection of the immunoblots

control variables

Total lysate, cytoplasmic and nucleoplasmic fractions
Secondary antibody (HRP-coupled anti-rabbit IgG)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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