Immunofluorescence Staining Assay for Spindle Proteins

Mouse anti-hMps1-N1 (Abcam, Ab11108, 1:500), mouse anti-Mad2 (CM2²⁷⁶, Santa Cruz, Sc-65492, 1:200), mouse anti-α-tubulin (Cell Signaling Technology, DM1A, 3873, 1:5000), mouse anti-MBP (Cell Signaling Technology, 8G1, 2396, 1:2000), rabbit anti-GFP (Proteintech, 50430-2-AP, 1:1000), mouse anti-His-tag (Cell Signaling Technology, 27E8, 2366, 1:2000), mouse anti-Flag (Sigma, F3165, 1:2000), and human anti-centromere auto-antibody (ACA, Immunovision, HCT-0100, 1:5000) were obtained commercially. Anti-pMELT-Knl1 antibody was kindly gifted by Dr Geert Kops (Hubrecht Institute, the Netherlands) (Nijenhuis et al., 2014 (link)).

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Gui P., Sedzro D.M., Yuan X., Liu S., Hei M., Tian W., Zohbi N., Wang F., Yao Y., Aikhionbare F.O., Gao X., Wang D., Yao X, & Dou Z. (2020). Mps1 dimerization and multisite interactions with Ndc80 complex enable responsive spindle assembly checkpoint signaling. Journal of Molecular Cell Biology, 12(7), 486-498.

Publication 2020

mouse anti-α-tubulin rabbit anti-GFP mouse anti-His-tag mouse anti-Flag

Anti antibody Anti centromere antibody Human Mouse Rabbit α tubulin

Corresponding Organization : Hefei National Center for Physical Sciences at Nanoscale

Other organizations : Morehouse School of Medicine, Huazhong University of Science and Technology, Zhejiang University

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Variable analysis

independent variables

Mouse anti-hMps1-N1 (Abcam, Ab11108, 1:500)
Mouse anti-Mad2 (CM2^276, Santa Cruz, Sc-65492, 1:200)
Mouse anti-α-tubulin (Cell Signaling Technology, DM1A, 3873, 1:5000)
Mouse anti-MBP (Cell Signaling Technology, 8G1, 2396, 1:2000)
Rabbit anti-GFP (Proteintech, 50430-2-AP, 1:1000)
Mouse anti-His-tag (Cell Signaling Technology, 27E8, 2366, 1:2000)
Mouse anti-Flag (Sigma, F3165, 1:2000)
Human anti-centromere auto-antibody (ACA, Immunovision, HCT-0100, 1:5000)
Anti-pMELT-Knl1 antibody (kindly gifted by Dr Geert Kops, Hubrecht Institute, the Netherlands)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: Anti-pMELT-Knl1 antibody (kindly gifted by Dr Geert Kops, Hubrecht Institute, the Netherlands)
Negative controls: Not specified

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