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TACC3 is a protein-coding gene that provides instructions for making the TACC3 protein. The TACC3 protein is involved in the process of cell division and is important for the proper separation of chromosomes during this process.

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Lab products found in correlation

Fgfr3, by Santa Cruz Biotechnology (2 mentions) Gm130, by BD (2 mentions) Myc tag, by Abcam (2 mentions) Pp2ac methyl leu 309, by BioLegend (2 mentions) Trappc2l, by Merck Group (2 mentions) Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Amersham ecl prime western blotting detection reagent, by GE Healthcare (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Non fat milk, by Bio-Rad (1 mentions) Alkbh5, by Abcam (1 mentions) Alkbh5, by Merck Group (1 mentions) C myc, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Vinculin, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Amersham ecl prime, by Cytiva (1 mentions) Nupage system, by Thermo Fisher Scientific (1 mentions) Phosphorylated akt, by Cell Signaling Technology (1 mentions) Phosphorylated stat3, by Cell Signaling Technology (1 mentions) Stat3, by Cell Signaling Technology (1 mentions)

5 protocols using tacc3

1

Western Blot Analysis of Protein Targets

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Cells were washed twice with ice-cold phosphate-buffered saline (PBS) and ruptured with RIPA buffer (Pierce, Rockford, IL) containing 5 mM EDTA, PMSF, cocktail proteinase inhibitors, and phosphatase inhibitor cocktail. Cell extracts were centrifuged at 12000 × g for 15 min and the supernatants were then collected. Cell lysates were resolved by SDS-PAGE and transferred onto PVDF membranes which were blocked with 5% non-fat milk (Bio-Rad) in Tris-buffered saline containing 0.1% Tween 20 for 1 hour and incubated sequentially with primary and secondary antibodies and detected by immunoblotting with the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) or Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare). Antibodies used for Western blotting were as follows: ALKBH5 (ab195377, abcam), ALKBH5 (HPA007196, Sigma-Aldrich), TACC3 (sc-376883, Santa Cruz Biotechnology), Flag (F3165, Sigma-Aldrich), c-Myc (9402S, Cell Signaling Technology), P21 (2947S, Cell Signaling Technology) GAPDH (sc-47724, Santa Cruz Biotechnology), β-Actin (3700, Cell Signaling Technology), Vinculin (sc-25336, Santa Cruz Biotechnology). GAPDH or β-Actin or Vinculin was used as a loading control.
Shen C., Sheng Y., Zhu A., Robinson S., Jiang X., Dong L., Chen H., Su R., Yin Z., Li W., Deng X., Chen Y., Hu Y.C., Weng H., Huang H., Prince E., Cogle C.R., Sun M., Zhang B., Chen C.W., Marcucci G., He C., Qian Z, & Chen J. (2020). M6A demethylase ALKBH5 selectively promotes tumorigenesis and cancer stem cell self-renewal in acute myeloid leukemia. Cell stem cell, 27(1), 64-80.e9.
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2

Western Blot Analysis of Signaling Proteins

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Cells were lysed in TNE buffer (10 mM Tris [pH 7.5], 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid [EDTA]) supplemented with 1% NP-40, protease inhibitor cocktail, and phosphatase inhibitor. Samples were separated using a NuPAGE system (Thermo Fisher Scientific) on 4–12% Bis-Tris gels in MOPS-SDS buffer, and then transferred to a polyvinylidene difluoride membrane. Antibodies were purchased from the indicated suppliers: FGFR3 (sc-13121, Santa Cruz; dilution ratio 1:200), TACC3 (sc-22773, Santa Cruz; dilution ratio 1:200), phosphorylated ERK (#4370, Cell Signaling Technology; dilution ratio 1:1000), ERK (#4695, Cell Signaling Technology; dilution ratio 1:1000), phosphorylated-AKT (#4060, Cell Signaling Technology; dilution ratio 1:1000), AKT (#4691, Cell Signaling Technology; dilution ratio 1:1000), phosphorylated-STAT3 (#9145, Cell Signaling Technology; dilution ratio, 1:1000), STAT3 (#9139, Cell Signaling Technology; dilution ratio 1:1000), and actin (MAB1501R, Merck Millipore Headquarters; dilution ratio 1:1000). Blots were then incubated with either anti-mouse or anti-rabbit horseradish peroxidase-conjugated secondary antibodies (#115-035-166 and #111-035-144, Jackson Immuno Research; dilution ratio 1:10,000), and visualized by chemiluminescence. Images have been cropped for presentation. Full size images are presented in Supplementary Figures 8–14.
Tamura R., Yoshihara K., Saito T., Ishimura R., Martínez-Ledesma J.E., Xin H., Ishiguro T., Mori Y., Yamawaki K., Suda K., Sato S., Itamochi H., Motoyama T., Aoki Y., Okuda S., Casingal C.R., Nakaoka H., Inoue I., Verhaak R.G., Komatsu M, & Enomoto T. (2018). Novel therapeutic strategy for cervical cancer harboring FGFR3-TACC3 fusions. Oncogenesis, 7(1), 4.
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3

Quantifying Protein Expression in Cells

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Protein lysates were quantitated by Bradford Assay. Twenty micrograms of total protein was resolved with a 4%–15% gradient gel and protein levels measured by immunoblot using the following antibodies: FGFR3 (sc-13121; Santa Cruz Biotechnology, 1:250), TACC3 (sc-22773 Santa Cruz Biotechnology, 1:1000), and α-tubulin (1:10,000).
Parker Kerrigan B.C., Ledbetter D., Kronowitz M., Phillips L., Gumin J., Hossain A., Yang J., Mendt M., Singh S., Cogdell D., Ene C., Shpall E, & Lang F.F. (2020). RNAi technology targeting the FGFR3-TACC3 fusion breakpoint: an opportunity for precision medicine. Neuro-oncology Advances, 2(1), vdaa132.
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4

Comprehensive Protein Quantification Assay

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Following primary antibodies were used at the indicated dilutions: Myc tag (#ab32, 1:1,000, Abcam), PPP2R1A (#2041, 1:2,500, Cell Signaling Technology), B55α (#5689, 1:2,500, Cell Signaling Technology), B56α (#610615, 1:3000, BD Biosciences), B56δ (#5687, 1:2,500, Cell Signaling Technology), B56ε (#PA5‐17981, 1:2,000, Thermo Fisher Scientific), PP2AC Methyl (Leu 309) (#828801, 1:3,000, BioLegend), PP2AC (#05‐421, 1:4,000, Millipore/Merk), actin, (#sc‐47778 HRP, 1:5,000, Santa Cruz Biotechnology), TRAPPC2L (#HPA041714, 1:2,000, Sigma‐Aldrich), TACC3 (sc‐376883, 1:1,000, Santa Cruz Biotechnology), ROD1 (1:300, a kind gift from Reto Gassmann’s lab), GM‐130 (#610822, 1:500, BD Biosciences), Tubulin (#T9026, DM1A, 1:2,000, Sigma‐Aldrich).
Vit G., Duro J., Rajendraprasad G., Hertz E.P., Holland L.K., Weisser M.B., McEwan B.C., Lopez‐Mendez B., Sotelo‐Parrilla P., Jeyaprakash A.A., Montoya G., Mailand N., Maeda K., Kettenbach A., Barisic M, & Nilsson J. (2022). Chemogenetic profiling reveals PP2A‐independent cytotoxicity of proposed PP2A activators iHAP1 and DT‐061. The EMBO Journal, 41(14), e110611.
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5

Antibody Validation Protocol

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Following antibodies were used at the indicated dilutions: Myc tag (#ab32, 1:1000, Abcam), PP2A scaffold subunit (#2041, 1:2500, Cell Signaling Technology), PPP2R2A (#5689, 1:2500, Cell Signaling Technology), PPP2R5A (#610615, 1:3000, BD Biosciences), PPP2R5D (#5687, 1:2500, Cell Signaling Technology), PPP2R5E (#PA5-17981, 1:2000, Thermo Fisher Scientific), PP2AC Methyl (Leu 309) (#828801, 1:3000, BioLegend), actin, (#sc-47778 HRP, 1:5000, Santa Cruz Biotechnology), TRAPPC2L (#HPA041714, 1:2000, Sigma-aldrich), TACC3 (sc-376883, 1:1000, Santa Cruz Biotechnology), ROD1 (1:300, a kind gift from Reto Gassmann's lab), GM-130 (#610822, 1:500, BD Biosciences), Alexa Fluor 488 (#A32723, 1:1000, Thermo Fisher Scientific), Tubulin (#T9026, DM1A, 1:2000, Sigma-Aldrich).
Vit G., Duro J., Rajendraprasad G., Hertz E.P.T., Holland L.K., Weisser M., McEwan B.C., López‐Méndez B., Montoya G., Mailand N., Maeda K., Kettenbach A.N., Barišić M., & Nilsson J. (2021). Cellular toxicity of iHAP1 and DT-061 does not occur through PP2A-B56 targeting.
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