Immunoblot Analysis of Protein Targets

Immunoblots were carried out as described⁷¹. Briefly, rabbit polyclonal α-FLAG (Rockland, dilution 1:2000), α-mCherry (Biovision, dilution 1:10000), α-PilC^{74 (link)} (dilution 1:5000) antibodies were used together with horseradish-conjugated goat anti-rabbit immunoglobulin G (Sigma-Aldrich, dilution 1:10000) as secondary antibody. Blots were developed using Luminata Crescendo Western HRP Substrate (Millipore) and visualized using a LAS-4000 luminescent image analyzer (Fujifilm).

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Skotnicka D., Steinchen W., Szadkowski D., Cadby I.T., Lovering A.L., Bange G, & Søgaard-Andersen L. (2020). CdbA is a DNA-binding protein and c-di-GMP receptor important for nucleoid organization and segregation in Myxococcus xanthus. Nature Communications, 11, 1791.

Publication 2020

α-mCherry horseradish-conjugated goat anti-rabbit immunoglobulin G Luminata Crescendo Western HRP Substrate LAS-4000 luminescent image analyzer

Anti immunoglobulin Antibodies Antibody Dilution Goat Horseradish Immunoblots Luminescent Rabbit

Corresponding Organization :

Other organizations : Max Planck Institute for Terrestrial Microbiology, Loewe Center for Synthetic Microbiology, University of Birmingham, Philipps University of Marburg

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Variable analysis

independent variables

Antibody types (rabbit polyclonal α-FLAG, α-mCherry, α-PilC)
Antibody dilutions (1:2000, 1:10000, 1:5000)

dependent variables

Protein expression levels detected by immunoblots

control variables

Horseradish-conjugated goat anti-rabbit immunoglobulin G (Sigma-Aldrich, dilution 1:10000) as secondary antibody
Luminata Crescendo Western HRP Substrate (Millipore) for blot development
LAS-4000 luminescent image analyzer (Fujifilm) for visualization

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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