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Alpha tubulin

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Alpha-tubulin is a protein that is a major component of microtubules, which are part of the cytoskeleton in eukaryotic cells. Microtubules play a crucial role in cellular processes such as cell division, cell motility, and intracellular transport. Alpha-tubulin is a valuable tool for researchers studying these cellular functions.

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Lab products found in correlation

Nitrocellulose membrane, by Bio-Rad (17 mentions) Western lightning ecl, by PerkinElmer (16 mentions) Sigmafast protease inhibitor cocktail, by Merck Group (16 mentions) Chemostar imager, by Intas (16 mentions) Poinceau, by Merck Group (14 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions) β actin, by Merck Group (7 mentions) Cleaved caspase 3, by Cell Signaling Technology (5 mentions) Odyssey infrared imaging system, by LI COR (5 mentions) Alexa fluor 488, by Thermo Fisher Scientific (4 mentions) Gamma tubulin, by Merck Group (4 mentions) Flag tag (flag), by Merck Group (4 mentions) Flag m2, by Merck Group (4 mentions) Ripa buffer, by Thermo Fisher Scientific (4 mentions) Gapdh, by Merck Group (4 mentions) Cleaved parp, by Cell Signaling Technology (3 mentions) Image studio, by LI COR (3 mentions) Gsk3β, by Cell Signaling Technology (3 mentions) β actin, by Santa Cruz Biotechnology (3 mentions) Gapdh, by Abcam (3 mentions)

135 protocols using alpha tubulin

1

Standardized Western Blotting Protocol

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma, Taufkirchen, Germany). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in PBS. The following antibodies were used: alpha-Tubulin (Sigma, Taufkirchen, Germany), HOXB5 (Santa Cruz Biotechnology, Heidelberg, Germany), and BCL6 (Cell Signaling Technology, Danvers, MA, USA). For loading control, blots were reversibly stained with Poinceau (Sigma, Taufkirchen, Germany) and detection of alpha-Tubulin (TUBA) performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western Lightning ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Nagel S., Pommerenke C., Meyer C., Kaufmann M, & MacLeod R.A. (2023). Chromosomal Aberration t(14;17)(q32;q21) Simultaneously Activates HOXB5 and miR10a in Triple-Hit B-Cell Lymphoma. Biomedicines, 11(6), 1758.
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2

Western Blot Analysis of Signaling Proteins

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: MSX1 (R & D Systems), alpha-Tubulin (Sigma), PDGFRB (R & D Systems), phospho-PDGFRB (Aviva Systems Biology, Eching, Germany), NKX2-2 (Aviva Systems Biology) and PITX1 (Abnova, Taipei, Taiwan). For loading control blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany). PDGFD and BMP4 were quantified in the medium by ELISA using according Quantikine ELISA kits from R & D Systems. Samples were obtained by harvesting supernatants of 1x106 cells which were washed in PBS and subsequently incubated in 1 ml medium for 24 h.
Nagel S., Pommerenke C., MacLeod R.A., Meyer C., Kaufmann M., Fähnrich S, & Drexler H.G. (2019). Deregulated expression of NKL homeobox genes in T-cell lymphomas. Oncotarget, 10(35), 3227-3247.
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3

Western Blot Analysis of IRF4 Expression

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (Sigma) and IRF4 (NOVUS Biologicals, Colorado, USA). For loading control blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Nagel S., Pommerenke C., Meyer C., Kaufmann M., MacLeod R.A, & Drexler H.G. (2017). NKL homeobox gene MSX1 acts like a tumor suppressor in NK-cell leukemia. Oncotarget, 8(40), 66815-66832.
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4

Western Blot Analysis and FGF2 ELISA

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Western blots were generated by the semi-dry method. Proteins obtained from cell line lysates using SIGMAFast protease inhibitor cocktail (Sigma) were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (Sigma), OTX1 (Santa Cruz Biotechnology, Heidelberg, Germany), OTX2 (Abcam, Cambridge, UK), and ZHX1 (Santa Cruz Biotechnology). For loading control the blots were stained with Poinceau (Sigma) and then detection of alpha-Tubulin (TUBA) was performed. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Quantification of FGF2 protein in cell culture supernatants was performed as follows: log-phase cells were washed twice in PBS and cultivated in fresh medium at 1x106 cells per ml and supernatants harvested after 24 h. ELISA was performed using the Human FGF basic Quantikine ELISA Kit (R&D Systems) and an ELISA reader (Thermo Electron, Vantaa, Finland).
Nagel S., Ehrentraut S., Meyer C., Kaufmann M., Drexler H.G, & MacLeod R.A. (2015). Aberrantly Expressed OTX Homeobox Genes Deregulate B-Cell Differentiation in Hodgkin Lymphoma. PLoS ONE, 10(9), e0138416.
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5

Western Blot Protein Detection Protocol

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (T6199, Sigma), HMX2 (NBP1-91997, Novus Biologicals, Abingdon, UK), HMX3 (LS-B8011, LSBio, Eching, Germany), ERK (sc-94, Santa Cruz Biotechnology, Heidelberg, Germany), phosphor (P)-ERK (sc-7383, Santa Cruz Biotechnology), EPX (LS-C354350, LSBio), HTR7 (LS-C358892, LSBio). For loading controls blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Nagel S., Pommerenke C., Meyer C., MacLeod R.A, & Drexler H.G. (2020). Aberrant expression of NKL homeobox genes HMX2 and HMX3 interferes with cell differentiation in acute myeloid leukemia. PLoS ONE, 15(10), e0240120.
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6

Golgi Morphology and SMN Analysis

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Cells were washed in PBS and fixed for 10 minutes in 4% paraformaldehyde followed by 30 minutes blocking and permeabilization in 5% normal goat serum with 0.2% Triton-X 100. For Golgi morphology, cells were incubated over night at 4°C with anti GM130 (BD bioscience, 1:500) and alpha-tubulin (Sigma, 1:4000). Cells were washed three times and incubated in secondary antibody (Alexa-488 and Alexa 594 1:1000) for 1 hr at room temperature. Cells were mounted in Prolong Gold with DAPI to visualize nuclei. Golgi morphology was classified as normal, intermediate, diffuse or absent by blinded observers with at least 50 cells from three separate experiments, and statistical significance was determined by Chi2 analysis. For detection of SMN in COPA depleted NSC-34 cells, cells were plated on 18mm glass coverslips coated with PDL and cultured in DMEM/F12 with 1% FBS for 72 hours to induce neurite outgrowth. Cultures were then infected with 4×106 lentivirus particles for 48 hours. Alpha-COP knockdown was confirmed by Western Blot analysis. Cells were stained with BIII-tubulin (rabbit, 1:1000 Cell Signaling Technologies) to visualize neurites and growth cones, and Mansma2 (1:50) to visualize SMN. For cells transfected with YFP-KDELR-VSVGts045 cells were immunolabeled with alpha-tubulin (Sigma, 1:5000) and Alexa 594 (goat anti-mouse 1:1000).
Custer S., Foster J., Astroski J, & Androphy E. (2018). Abnormal Golgi morphology and decreased COPI function in cells with low levels of SMN. Brain research, 1706, 135-146.
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7

Western Blotting and IL-17F ELISA Protocol

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (T6199, Sigma), HLX (Novus Biologicals, Abingdon, UK), STAT3 (9139, Cell Signalling, Leiden, Netherlands) and phospho-STAT3 (9136, Cell Signalling). For loading controls blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
IL17F protein was quantified in cell culture supernatants using the IL-17 Human ELISA kit (Thermo Fisher Scientific). To harvest the supernatants 1 × 106 cells were washed in PBS and cultured in 1 ml fresh medium. After 24 hours supernatants were centrifuged and stored at –20°C. To optimize the range of the kit supernatants were diluted 1:50 and 1:100 in PBS.
Nagel S., Pommerenke C., MacLeod R.A., Meyer C., Kaufmann M, & Drexler H.G. (2020). The NKL-code for innate lymphoid cells reveals deregulated expression of NKL homeobox genes HHEX and HLX in anaplastic large cell lymphoma (ALCL). Oncotarget, 11(34), 3208-3226.
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8

Western Blotting and BMP2 ELISA Protocol

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma, Taufkirchen, Germany). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (Sigma, #T6199), IRX1 (Biozol, Eching, Germany, #DF3225), and IRX3 (Biozol, #MBS8223417). For loading control, blots were reversibly stained with Poinceau (Sigma) and the detection of alpha-Tubulin (TUBA) was performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
The enzyme-linked immunosorbent assay (ELISA) was used to quantify BMP2 protein levels in the supernatant of cell cultures. In addition, 2 × 106 cells were cultured in 2 mL of fresh medium in a 24-well plate. After 24 h, 1 of mL medium was harvested and frozen in aliquots. Quantification was performed using the Quantikine ELISA BMP-2 kit (R & D Systems, #DBP200), as described by the company. Two biological replicates were analyzed in triplicate.
Nagel S., Pommerenke C., Meyer C, & MacLeod R.A. (2022). The Hematopoietic TALE-Code Shows Normal Activity of IRX1 in Myeloid Progenitors and Reveals Ectopic Expression of IRX3 and IRX5 in Acute Myeloid Leukemia. International Journal of Molecular Sciences, 23(6), 3192.
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9

Western Blot Analysis of Cell Lysates

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Western blots were generated by the semi-dry method. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). Proteins were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (Sigma, #T6199), PBX1 (LifeSpan Biosciences, Eching, Germany, #LS-C133363), and NFIB (Novus Biologicals, Abingdon, UK, #NBP1-81000). For loading control blots were reversibly stained with Poinceau (Sigma) and detection of alpha-Tubulin (TUBA) performed thereafter. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Nagel S., Pommerenke C., Meyer C., MacLeod R.A, & Drexler H.G. (2021). Establishment of the TALE-code reveals aberrantly activated homeobox gene PBX1 in Hodgkin lymphoma. PLoS ONE, 16(2), e0246603.
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10

Western Blot Protein Detection Protocol

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Western blots were generated by the semi-dry method. Proteins obtained from cell line lysates using SIGMAFast protease inhibitor cocktail (Sigma) were transferred onto nitrocellulose membranes (Bio-Rad, München, Germany) and blocked with 5% dry milk powder dissolved in phosphate-buffered-saline buffer (PBS). The following antibodies were used: alpha-Tubulin (Sigma), and SIX1 (Novus Biologicals Europe, Cambridge, UK). For loading control the blots were stained with Poinceau (Sigma) and then detection of alpha-Tubulin (TUBA) was performed. Secondary antibodies were linked to peroxidase for detection by Western-Lightning-ECL (Perkin Elmer, Waltham, MA, USA). Documentation was performed using the digital system ChemoStar Imager (INTAS, Göttingen, Germany).
Nagel S., Meyer C., Kaufmann M., Drexler H.G, & MacLeod R.A. (2015). Aberrant expression of homeobox gene SIX1 in Hodgkin lymphoma. Oncotarget, 6(37), 40112-40126.
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