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Mouse monoclonal anti α tubulin clone dm1a

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Mouse monoclonal anti-α-tubulin (clone DM1A) is a laboratory reagent used for the detection and analysis of α-tubulin, a key structural component of microtubules. This antibody is produced in mice and can be used to visualize and study the distribution and dynamics of α-tubulin in various cell types and tissues.

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Lab products found in correlation

Peroxidase and, by Jackson ImmunoResearch (2 mentions) Anti aurora b, by Abcam (2 mentions) Mouse monoclonal anti flag clone m2, by Merck Group (2 mentions) Alexa fluor conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions) Mouse monoclonal anti cit k, by BD (2 mentions) Phospho drosophila p70 s6 kinase thr398, by Cell Signaling Technology (1 mentions) Phospho akt thr308, by Cell Signaling Technology (1 mentions) Rabbit polyclonal anti ha h6908, by Merck Group (1 mentions) Goat polyclonal anti egfp, by Rockland Immunochemicals (1 mentions) Adenosine 5 diphosphate, by Merck Group (1 mentions) Alexa fluor conjugated antibody, by Thermo Fisher Scientific (1 mentions) Poly l lysine solution, by Merck Group (1 mentions) Hoechst 33258, by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Human fibronectin, by BD (1 mentions) Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Proteasome inhibitor cocktail, by Merck Group (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Anti mouse igg dylight680, by Thermo Fisher Scientific (1 mentions)

12 protocols using mouse monoclonal anti α tubulin clone dm1a

1

Immunoblotting Analysis of Signaling Proteins

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The following antibodies were used: rabbit polyclonal anti-FMRP (Sigma, F4055), mouse monoclonal anti-αTubulin (clone DM1A, Sigma), rabbit polyclonal anti-p110β (Millipore, 09–482), rabbit polyclonal anti-PIKE-L/S (Millipore, 07–675), rabbit polyclonal anti-mGlu5 (Millipore, AB5675), mouse monoclonal anti-dFMR1 (clone 6A15, Abcam), and mouse monoclonal anti-fasciclin II (clone 1D4, NeuroMab). The following rabbit monoclonal antibodies from Cell Signaling Technology were used: phospho-Akt(Thr308) (#4056), phospho-Drosophila p70 S6 Kinase (Thr398) (#9209), IRS-2 (#4502).
Gross C., Chang C.W., Kelly S.M., Bhattacharya A., McBride S.M., Danielson S.W., Jiang M.Q., Chan C.B., Ye K., Gibson J.R., Klann E., Jongens T.A., Moberg K.H., Huber K.M, & Bassell G.J. (2015). Increased expression of the PI3K enhancer PIKE mediates deficits in synaptic plasticity and behavior in Fragile X syndrome. Cell reports, 11(5), 727-736.
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2

Antibody Sources and Validation

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The following antibodies were obtained from the indicated commercial sources: mouse monoclonal anti-HA (clone HA-7; Covance), rabbit polyclonal anti-HA (H6908; Sigma), mouse monoclonal anti-α-tubulin (clone DM1A; Sigma), rabbit polyclonal anti-actin (A2066; Sigma), goat polyclonal anti-EGFP (Rockland), and mouse monoclonal anti-GM130 (clone 35/GM130; BD Transduction Laboratories). The anti-synapsin antibody was previously described (Han et al., 2016b (link)).
Kang H., Han K.A., Won S.Y., Kim H.M., Lee Y.H., Ko J, & Um J.W. (2016). Slitrk Missense Mutations Associated with Neuropsychiatric Disorders Distinctively Impair Slitrk Trafficking and Synapse Formation. Frontiers in Molecular Neuroscience, 9, 104.
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3

Platelet Function Assays Protocol

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Adenosine 5′-diphosphate (ADP), Hoechst 33258, Poly-L-lysine solution, paraformaldehyde (PFA), eosin-hematoxylin solution, Triton X-100, tetramethyrhodamine isothiocyanate (TRITC)-conjugated phalloidin and cyclopiazonic acid from Penicillium cyclopium (CPA) were from Sigma Aldrich (Milan, Italy). Fura-2 acetoxymethyl ester (Fura-2 AM) was from Molecular Probes Europe BV (Leiden, The Netherlands). Human fibronectin was from BD Bioscience (Milan, Italy). Type I collagen was purified as described previously35 (link). The following antibodies were used: monoclonal anti-CD61 (clone SZ21) (Immunotech, Marseille, France); goat monoclonal anti-CD61 (clone C-20) and mouse monoclonal anti-P-Selectin (clone 1E3) (Santa Cruz Biotechnology, California, USA); mouse monoclonal anti-α-tubulin (clone DM1A) and mouse monoclonal anti-human P-Selectin-allophycocyanin (APC) (clone clone AK4) (Sigma Aldrich, Milan, Italy); rabbit polyclonal anti-von Willebrand Factor (Dako, Milan, Italy); mouse monoclonal anti-Thrombospondin (clone A6.1), mouse monoclonal anti-CD61-fluorescein isothiocyanate (FITC) (clone PM6/13) and mouse monoclonal anti-CD42b-phycoerythrin (PE) (clone HIP1) (Abcam, Cambridge, UK); anti-CD61-FITC and anti-CD42b-PE (BD Biosciences, San José, CA, USA); Alexa Fluor-conjugated antibodies (Invitrogen, Milan, Italy).
Di Buduo C.A., Alberelli M.A., Glembostky A.C., Podda G., Lev P.R., Cattaneo M., Landolfi R., Heller P.G., Balduini A, & De Candia E. (2016). Abnormal proplatelet formation and emperipolesis in cultured human megakaryocytes from gray platelet syndrome patients. Scientific Reports, 6, 23213.
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4

Generation and Characterization of CHMP4C Phospho-Specific Antibodies

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CHMP4C phospho-specific antibodies were raised in rabbits against synthetic CHMP4C peptides encompassing residues 206–220 (TARRSRAASSQRAEEC) and containing either a phosphorylated serine at position S210 (TARRpSRAASSQRAEEC; mono-phospho CHMP4C) or phosphorylated serines at position S210, S214 and S215 (TARRpSRAApSpSQRAEEC; triphospho CHMP4C). Sera were first eluted through a column containing a non-phosphorylated peptide, and then each antibody was affinity purified using the appropriate phosphopeptide. Peptide synthesis, conjugation, rabbit immunizations, serum production and affinity purification were carried out by Generon, UK.
Other antibodies used in this study were: mouse monoclonal anti α-tubulin (clone DM1A, Sigma, T9026), chicken polyclonal anti-α-tubulin (Abcam, ab89984), mouse monoclonal anti-Flag (clone M2, Sigma, F3165), rabbit polyclonal anti-Aurora B (Abcam, ab2254), mouse monoclonal anti-Borealin (clone 1D11-MLB Life Science M147-3), rabbit polyclonal anti-KIF23/MKLP1 (clone N19, Santa Cruz Biotechnology, sc-867), rabbit polyclonal anti-CHMP4B, a gift of Sagona & Stenmark [36 (link)]. Peroxidase- and Alexa-fluor-conjugated secondary antibodies were purchased from Jackson Laboratories and Thermo, respectively.
Capalbo L., Mela I., Abad M.A., Jeyaprakash A.A., Edwardson J.M, & D'Avino P.P. (2016). Coordinated regulation of the ESCRT-III component CHMP4C by the chromosomal passenger complex and centralspindlin during cytokinesis. Open Biology, 6(10), 160248.
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5

Immunodetection of Apoptosis Signaling

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The following antibodies were used: mouse monoclonal anti-α-tubulin (clone DM1A, Sigma, T9026), mouse monoclonal anti-CIT-K (BD Transduction Laboratories, 611377), rabbit anti-cleaved caspase 3 (Cell Signaling Technology, CST, 9661), rabbit anti-cleaved caspase 6 (CST, 9761), rabbit anti-cleaved caspase 7 (CST, 9491), rabbit anti-cleaved PARP (CST, 9541), mouse monoclonal anti-p53 (clone D0-1, Santa Cruz Biotechnology, sc-126), rabbit anti-phosphorylated LATS (CST, D57D3), rabbit anti-LATS2 (Bethyl Laboratories, A300-479A) and mouse monoclonal anti-TUBB3 (clone 2G10, Abcam, ab78078). Peroxidase and Alexa-fluor conjugated secondary antibodies were purchased from Jackson Laboratories and Invitrogen respectively.
McKenzie C, & Paolo D’Avino P. (2016). Investigating cytokinesis failure as a strategy in cancer therapy. Oncotarget, 7(52), 87323-87341.
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6

Quantifying Cellular BDNF Protein Levels

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Protein expression was assessed by Western blotting. Cells were harvested from the culture plates and total cellular proteins were extracted by lysis buffer containing 1% Triton X-100, 1% proteasome inhibitor cocktail (Sigma-Aldrich). Lysates were cleared by centrifugation at 13,000 g for 20 min and protein concentrations were measured using the BC Assay Protein Quantitation Kit (Uptima, Oakland, CA). Total protein lysates (50 μg) were fractionated on 15% SDS-PAGE then transferred to nitrocellulose membranes (GE Healthcare Life Sciences, Vélizy-Villacoublay, France). Membranes were blocked for 1 h in PBST with 2.5% BSA, and then incubated with primary antibodies overnight at 4 °C, rabbit polyclonal anti-BDNF (sc-546, Santa Cruz [51 (link)]) at 1:200 and mouse monoclonal anti-α-tubulin (Clone DM1A, Sigma-Aldrich) at 1:10,000. Incubation with secondary antibodies conjugated to infrared fluorophores (goat anti-rabbit IgG Dylight 800 and anti-mouse IgG Dylight 680 at 1:10,000 from Thermo Scientific) was performed for 1 h. An Odyssey infrared imaging system (LI-COR, Bad Homburg, Germany) was used to scan membranes at a wavelength of 680 nm (anti-mouse) or 800 nm (anti-rabbit). Data were analyzed with Image Studio 1.1 software (Li-COR).
Chen H., Lombès M, & Le Menuet D. (2017). Glucocorticoid receptor represses brain-derived neurotrophic factor expression in neuron-like cells. Molecular Brain, 10, 12.
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7

Antibodies and Selective Inhibitors for Cell Signaling

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The following primary antibodies were used: Rabbit polyclonal anti-SNX18 IgG (AbCam, ab99035); mouse monoclonal anti-myc (Cell Signaling Technology, 2272); Rabbit polyclonal anti-β-Tubulin (LI-COR Biosciences, 926-42211); mouse monoclonal anti-α-tubulin Clone DM 1A (Sigma Aldrich, T9026), anti-α1 Na+/K+ ATPase antibody (ab7671, Abcam), mouse monoclonal anti-LPS (Abcam, ab 8274); Mouse anti-Phospho-Tyrosine antibody, clone 4G10 (Millipore, 05-1050); mouse monoclonal anti-HA (Covance); Rabbit monoclonal anti-Akt (pan) C67E7 (Cell Signaling, 4691); and anti-phospho-Akt (Ser473) D9E (Cell Signaling, 4060). Goat anti-mouse coupled to Alexa Fluor 405, 546 or 647 (Invitrogen) were used as secondary antibodies for immunofluorescence. The IRDye800CW goat anti-mouse IgG; IRDye800CW donkey anti-rabbit IgG; IRDye680LT goat anti-rabbit IgG; and IRDye680LT donkey anti-mouse IgG were purchased from LI-COR Biosciences. Phalloidin-Alexa Fluor 635, fixable analog of lipophilic membrane stain FM 4-64FX and Wheat germ agglutinin (WGA) coupled to Alexa Fluor 647 were from Invitrogen (A34054, F34653, and W324666). Selective inhibitor of Rac1-GEF interaction NSC 23766 was purchased from Tocris Bioscience and the Akt1/2 kinase inhibitor from Sigma Aldrich.
Liebl D., Qi X., Zhe Y., Barnett T.C, & Teasdale R.D. (2017). SopB-Mediated Recruitment of SNX18 Facilitates Salmonella Typhimurium Internalization by the Host Cell. Frontiers in Cellular and Infection Microbiology, 7, 257.
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8

Antibody Sources for Signaling Pathways

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Antibodies were purchased from the following resources: mouse monoclonal anti-Cathepsin K (clone 182-12G5) (Millipore); rat anti-Notch2 (clone C651.6DbHN) (Developmental Studies Hybridoma Bank); rabbit polyclonal anti-ERK1/2, mouse monoclonal anti-phospho-ERK1/2 (Thr202/Tyr204), mouse monoclonal anti-Akt (pan) (clone 40D4), rabbit monoclonal anti-phosphor-Akt (Ser473) (clone 193H12), rabbit polyclonal anti-JNK, mouse monoclonal anti-phospho-JNK (Thr183/Tyr185) (clone G9), rabbit polyclonal anti-IKB-α, mouse monoclonal anti-phospho-IKB-α (Ser32/36) (clone 5A5), rabbit monoclonal anti-cleaved caspase-3 (catalog number 9664), rabbit polyclonal anti-PARP (catalog number 9542), rabbit monoclonal anti-Numb (C29G11) (catalog number 2756) (Cell Signaling Technology); mouse monoclonal anti-α-tubulin (clone DM1A) (Sigma-Aldrich); mouse monoclonal anti-beta actin (catalog number A00702) (GenScript).
Cell culture alpha-MEM and 10 × Penicillin-Streptomycin-L-Glutamine were purchased from Life Technologies and Sigma-Aldrich, respectively. Fetal bovine serum was purchased from Hyclone.
Zhou J., Fujiwara T., Ye S., Li X, & Zhao H. (2015). Ubiquitin E3 Ligase LNX2 is Critical for Osteoclastogenesis in vitro by Regulating M-CSF/RANKL Signaling and Notch2. Calcified tissue international, 96(5), 465-475.
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9

Immunofluorescence Microscopy of Cell Markers

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The following primary antibodies were used for immunofluorescence microscopy: mouse monoclonal anti-E-cadherin, clone 36 (BD Transduction Laboratories); mouse monoclonal anti-β-actin, clone 4C2 (Abcam); mouse monoclonal anti-α-tubulin, clone DM1A (Sigma); mouse monoclonal anti-zyxin, clone 164D4 (Synaptic Systems); rabbit polyclonal anti-EPLIN (Novus Biologicals NB100-2305, lot A4); mouse monoclonal anti-β-catenin, clone 14 (BD Transduction Laboratories); rabbit polyclonal anti-p34-Arc/ARPC2 (Upstate, Merck), rabbit polyclonal anti-phospho-p44/42 MAPK (ERK1/2) (Cell Signaling Technology). The following secondary antibodies from Jackson ImmunoResearch were used: goat polyclonal anti-mouse IgG, IgG1, or IgG2a, or anti-rabbit IgG conjugated with AlexaFluor488, AlexaFluor594, or AlexaFluor647. AlexaFluor488-conjugated phalloidin (Molecular Probes) or TRITC-conjugated phalloidin (Fluka) were added to secondary antibodies. Horseradish peroxidase-conjugated goat polyclonal anti-mouse and anti-rabbit IgG antibodies (Jackson ImmunoResearch) were used for Western blot analysis. Other reagents were obtained from Sigma.
Zhitnyak I.Y., Rubtsova S.N., Litovka N.I, & Gloushankova N.A. (2020). Early Events in Actin Cytoskeleton Dynamics and E-Cadherin-Mediated Cell-Cell Adhesion during Epithelial-Mesenchymal Transition. Cells, 9(3), 578.
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10

Immunofluorescence Staining of Cellular Structures

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DNA-binding probe DAPI (4,6-Diamino-2-phenylindole dyhydrochloride) was obtained from Molecular Probes (USA). Mouse monoclonal anti-flotillin-2 antibody (clone 29) was from BD Transduction Labs (USA). Rabbit polyclonal anti-chick desmin antibody was a gift from Dr. Howard Holtzer (University of Pennsylvania, USA). Rabbit anti-chick MyoD antibody [10] (link) was kindly provided by Dr. Bruce Paterson (National Cancer Institute, NIH, USA). Mouse monoclonal anti-α-tubulin (clone DM1A) was from Sigma-Aldrich (USA). Alexa Fluor 488-goat anti-mouse/rabbit IgG and Alexa Fluor 546-goat anti-mouse/rabbit IgG antibodies were from Molecular Probes (USA). Peroxidase-conjugated goat anti-mouse/rabbit antibodies were obtained from Amersham Biosciences (UK). Goat anti-mouse 10-ηm gold-conjugated antibody was from BB International (USA).
Possidonio A.C., Soares C.P., Portilho D.M., Midlej V., Benchimol M., Butler-Browne G., Costa M.L, & Mermelstein C. (2014). Differences in the Expression and Distribution of Flotillin-2 in Chick, Mice and Human Muscle Cells. PLoS ONE, 9(8), e103990.
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