Antibody Production and Characterization

Antiserum against a peptide corresponding to amino acids 25–39 of human CHMP6 was raised in two rabbits and affinity purified on Affi-Gel beads containing immobilized peptide. Rabbit anti-CHMP3, rabbit anti-ALIX, rabbit anti-EAP30, rabbit anti-CHMP4B, and rabbit anti-EAP20 were described previously (Sharma et al., 2004 (link); Cabezas et al., 2005 (link); Bache et al., 2006 (link); Malerød et al., 2007 (link); Sagona et al., 2010 (link)). Mouse anti–α-tubulin (Sigma-Aldrich), sheep anti–α-tubulin (Cytoskeleton), mouse anti–β-actin (Sigma-Aldrich), goat anti-RacGAP1 (Abcam), rabbit anti-MKLP1 (Santa Cruz Biotechnology, Inc.), goat anti-V5 (Abcam), mouse anti-GFP (Roche), mouse anti-TSG101 (BD Transduction Laboratories), rabbit anti-CHMP6 (Santa Cruz Biotechnology, Inc.), rabbit anti-CEP55 (Abnova), rabbit anti-VPS28 (Santa Cruz Biotechnology, Inc.), rabbit anti-IST1 (Proteintech), rabbit anti-CHMP4A (Santa Cruz Biotechnology, Inc.), and rabbit anti-mCherry (Acris) were used as primary antibodies. Secondary antibodies included anti-mouse, anti-rabbit, and anti-goat Alexa Fluor 488 (Jackson ImmunoResearch), Alexa Fluor 555 (Molecular Probes), Alexa Fluor 568 (Molecular Probes), Alexa Fluor 647 (Jackson ImmunoResearch), and DyLight649 (Jackson ImmunoResearch).

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Christ L., Wenzel E.M., Liestøl K., Raiborg C., Campsteijn C, & Stenmark H. (2016). ALIX and ESCRT-I/II function as parallel ESCRT-III recruiters in cytokinetic abscission. The Journal of Cell Biology, 212(5), 499-513.

Publication 2016

Mouse anti–α-tubulin mouse anti–β-actin mouse anti-GFP mouse anti-TSG101 rabbit anti-VPS28 Alexa Fluor 488 Alexa Fluor 555 Alexa Fluor 568 Alexa Fluor 647 DyLight649

Actin Alexa 568 Alexa fluor 488 Alexa fluor 555 Alexa fluor 647 Amino acids Antibodies Antiserum Cytoskeleton Goat Human Molecular probes Mouse Peptide Rabbit Rabbits Sheep Tsg101 α tubulin

Corresponding Organization :

Other organizations : University of Oslo, Oslo University Hospital

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Variable analysis

independent variables

Antiserum against a peptide corresponding to amino acids 25–39 of human CHMP6 was raised in two rabbits and affinity purified on Affi-Gel beads containing immobilized peptide.

dependent variables

Not explicitly mentioned

control variables

Rabbit anti-CHMP3, rabbit anti-ALIX, rabbit anti-EAP30, rabbit anti-CHMP4B, and rabbit anti-EAP20 were described previously (Sharma et al., 2004; Cabezas et al., 2005; Bache et al., 2006; Malerød et al., 2007; Sagona et al., 2010).
Mouse anti–α-tubulin (Sigma-Aldrich), sheep anti–α-tubulin (Cytoskeleton), mouse anti–β-actin (Sigma-Aldrich), goat anti-RacGAP1 (Abcam), rabbit anti-MKLP1 (Santa Cruz Biotechnology, Inc.), goat anti-V5 (Abcam), mouse anti-GFP (Roche), mouse anti-TSG101 (BD Transduction Laboratories), rabbit anti-CHMP6 (Santa Cruz Biotechnology, Inc.), rabbit anti-CEP55 (Abnova), rabbit anti-VPS28 (Santa Cruz Biotechnology, Inc.), rabbit anti-IST1 (Proteintech), rabbit anti-CHMP4A (Santa Cruz Biotechnology, Inc.), and rabbit anti-mCherry (Acris) were used as primary antibodies.

controls

Positive controls: Rabbit anti-CHMP3, rabbit anti-ALIX, rabbit anti-EAP30, rabbit anti-CHMP4B, and rabbit anti-EAP20 were described previously.
Negative controls: Not explicitly mentioned

Annotations

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