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Mklp1

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MKLP1 is a molecular motor protein that plays a role in the separation of chromosomes during cell division. It is involved in the transportation and organization of chromosomes during mitosis.

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Lab products found in correlation

β tubulin, by Merck Group (2 mentions) Flag tag (flag), by Merck Group (1 mentions) α tubulin, by Abcam (1 mentions) Tsg101, by GeneTex (1 mentions) α tubulin, by Merck Group (1 mentions) γ tubulin, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Centrin 2, by Santa Cruz Biotechnology (1 mentions) Cdkl5, by Merck Group (1 mentions) Cep170, by Thermo Fisher Scientific (1 mentions) Phospho h3s10, by Abcam (1 mentions) Irdye 800 goat anti mouse igg, by LI COR (1 mentions) α actinin, by Abcam (1 mentions) Hsp90, by BD (1 mentions) Tsg101, by BD (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) β tubulin, by Cell Signaling Technology (1 mentions) Rab2a, by Thermo Fisher Scientific (1 mentions) Irdye 700 goat anti rabbit igg, by LI COR (1 mentions) Racgap1, by Santa Cruz Biotechnology (1 mentions)

3 protocols using mklp1

1

Antibody Specific to Phospho-CEP55 (S436)

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Antibody specific to phospho-CEP55 (S436) was raised against the synthetic phosphopeptide CKRIFKDLGTPFAL(pS)KSSM. Phospho-specific antibody was obtained though two-step affinity-purification methods. Commercially available antibodies for WB were as follows: ASPP1 (ab137537; Abcam), ASPP2 (611354; BD Biosciences), iASPP (18590-1-AP; Proteintech), CEP55 (23891-1-AP; Proteintech), CEP55 (7825-1; epitomics), α-Tubulin (1878-s; epitomics), β-Tubulin (05–661; Millipore), ALIX (sc-67338; Santa Cruz), TSG101 (4A10; Genetex), CHMP2B (12527-1-AP; Proteintech), CHMP4B(13683-1-AP; Proteintech), MKLP-1 (sc-867; Santa Cruz), MgcRacGAP (H00029127; Novus), PP1α (1950–1; epitomics), PP1β (2029–1; epitomics), PP1γ (6646–1; epitomics), Myc (9E10; Sigma), FLAG (M2; Sigma), HA (MM5-101R; Millipore), and Actin (AC-74; Sigma).
Gao K., Zhang Y., Shi Q., Zhang J., Zhang L., Sun H., Jiao D., Zhao X., Tao H., Wei Y., Wang Y., Saiyin H., Zhao S.M., Li Y., Zhang P, & Wang C. (2018). iASPP–PP1 complex is required for cytokinetic abscission by controlling CEP55 dephosphorylation. Cell Death & Disease, 9(5), 528.
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2

Antibody Detection Protocol for Cellular Proteins

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Antibodies (Abs) against the following proteins were used: β-actin (Sigma-Aldrich, St Louis, MO, USA, A5441), α-tubulin (Sigma-Aldrich, T6074), β-tubulin (Sigma-Aldrich, C4585), γ-tubulin (Sigma-Aldrich, T5326), CDKL5 (polyclonal, Sigma-Aldrich, HPA002847; monoclonal Santa Cruz, clone D-12, sc-376314), AKT (Cell Signaling, 4685), Aurora kinase A (Sigma-Aldrich, A1231, clone 35C1), GFP (Roche Diagnostics Ltd, 1814460), PLK-1 (Santa Cruz, sc-17783), phospho-H3-S10 (Abcam, ab14955), H2SB14-P (Cell Signaling, 6959); centrin-2 (Santa Cruz, sc-27793), Cep170 (ThermoFisher Scientific, 41-3200) and HIPK2 (rabbit polyclonal21 (link)); MgcRacGAP1 (Abcam, ab2270), MKLP1 (Santa Cruz; sc-22793); PRC1 (Santa Cruz, Sc-8356); spastin (Santa Cruz, Sc-53443), CREST centromere protein (Antibodies Inc., 15-234-0001). HRP-conjugated goat anti-mouse or anti-rabbit secondary Abs for immunoblotting were purchased from Thermo Fisher Scientific. DAPI and secondary Alexa Fluor anti-rabbit, anti-mouse, anti-human, and anti-goat Abs for immunofluorescence were obtained from Life Technologies while FITC- or TRITC-conjugated Abs were from Jackson Immuno Research Lab.
Barbiero I., Valente D., Chandola C., Magi F., Bergo A., Monteonofrio L., Tramarin M., Fazzari M., Soddu S., Landsberger N., Rinaldo C, & Kilstrup-Nielsen C. (2017). CDKL5 localizes at the centrosome and midbody and is required for faithful cell division. Scientific Reports, 7, 6228.
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3

Protein Quantification and Immunoblotting

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Protein samples were quantified by 1D SDS-PAGE/SYPRO Ruby protein staining-based densitometry18 (link). Western blotting was performed on protein samples (10–20 µg) as previously described18 (link). Rabbit antibodies raised against GAPDH (Cell Signalling), β-tubulin (Cell Signalling), KRASG12V mutant specific (Cell Signalling), RAB7 (Abcam) and GFP (Abcam) were used. Mouse antibodies against MKLP1 (Santa Cruz), RACGAP1 (Santa Cruz), ALIX (BD Biosciences), TSG101 (BD Biosciences), CD63 (Santa Cruz), CD81 (Santa Cruz), RAB2A (Thermo Fisher), FLOT1 (BD Biosciences), α-actinin (Abcam), CD9 (Santa Cruz) and HSP90 (BD Biosciences) were used. Secondary antibodies used were IRDye 800 goat anti-mouse IgG or IRDye 700 goat anti-rabbit IgG (1:15000, LI-COR Biosciences).
Rai A., Greening D.W., Xu R., Chen M., Suwakulsiri W, & Simpson R.J. (2021). Secreted midbody remnants are a class of extracellular vesicles molecularly distinct from exosomes and microparticles. Communications Biology, 4, 400.
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