Indirect Immunofluorescence for Tubulin and Sgo1-6HA

Indirect immunofluorescence was performed as described in Visintin et al. (1999) (link). Tubulin was visualized using a rat anti-tubulin antibody (AbD Serotec) at a dilution of 1:50 and an anti-rat FITC-conjugated antibody (Jackson ImmunoResearch) at a dilution of 1:100. For detection of Sgo1-6HA, a mouse HA.11 antibody (Covance) at a dilution of 1:500 and an anti-mouse Cy3-conjugated antibody (Jackson ImmunoResearch) at a dilution of 1:100 were used. Chromosome spreads were performed as described in Bizzari and Marston (2011) (link).

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Nerusheva O.O., Galander S., Fernius J., Kelly D, & Marston A.L. (2014). Tension-dependent removal of pericentromeric shugoshin is an indicator of sister chromosome biorientation. Genes & Development, 28(12), 1291-1309.

Publication 2014

rat anti-tubulin antibody anti-rat FITC-conjugated antibody HA.11 antibody anti-mouse Cy3-conjugated antibody

Anti antibody Antibody Chromosome Dilution Fitc Indirect immunofluorescence Mouse Sgo1 Tubulin

Corresponding Organization :

Other organizations : Wellcome Centre for Cell Biology, University of Edinburgh

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Variable analysis

independent variables

Indirect immunofluorescence protocol (as described in Visintin et al. (1999))
Primary antibodies used: rat anti-tubulin antibody (1:50 dilution) and mouse HA.11 antibody (1:500 dilution)
Secondary antibodies used: anti-rat FITC-conjugated antibody (1:100 dilution) and anti-mouse Cy3-conjugated antibody (1:100 dilution)

dependent variables

Visualization of tubulin
Detection of Sgo1-6HA

control variables

Chromosome spreads performed as described in Bizzari and Marston (2011)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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