alpha Tubulin Antibody (DM1A)

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - Staining of skin fibroblasts.

Simple Western: alpha Tubulin Antibody (DM1A) [NB100-690] - Simple Western lane view shows a specific band for alpha Tubulin in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Alpha tubulin molecular weight: 50 kDa.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of formalin fixed colon sections. Heat mediated antigen retrieval was performed using sodium citrate buffer for 20 min before incubating with primary antibody at a 0.5ug/ml dilution for 15 min at RT.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of colon tissue. Sections were formalin fixed and embedded with paraffin. Sodium citrate heat mediated antigen retrieval for 20 min. Incubated with primary antibody for 15 min at a 5 ug/ml concentration. Corner image is staining with secondary only.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of formalin fixed paraffin embedded heart sections. Used at a dilution of 1:500.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analsis of alpha tubulin in 9 cell lysates. Lane 1. HeLa; Lane 2. JURKAT; Lane 3. COS7; Lane 4. NIH-3T3; Lane 5. PC-12; Lane 6. RAT2; Lane 7. CHO; Lane 8. MDBK; Lane 9. MDCK

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using alpha Tubulin antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Immunomicroscopy: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of HeLa cells, green staining is alpha tubulin whereas red is DNA stained with propidium iodide.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of HeLa and COS-7 lysates. Alpha tubulin molecular weight: 50 kDa.

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-alpha Tubulin (DM1A) (NB100-690) at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse prostate using alpha Tubulin Antibody (DM1A) at 1:200 dilution. The signal was developed using HRP labelled secondary and DAB reagent which followed counterstaining with hematoxylin. The antibody generated a specific cytoplasmic/cytoskeletal staining in the prostate epithelial cells.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin (DM1A) Antibody NB100-690JF646 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Janelia Fluor 646.

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Western blot analysis of extracts from HeLa, COS and C6 cells using alpha Tubulin antibody (NB100-690, 1:1000, Alpha tubulin molecular weight: 50 kDa)

Western Blot: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of alpha tubulin (molecular weight of 50 kDa) in 9 cell lysates. Lane 1. HeLa; Lane 2. JURKAT; Lane 3. COS7; Lane 4. NIH-3T3; Lane 5. PC-12; Lane 6. RAT2; Lane 7. CHO; Lane 8. MDBK; Lane 9. MDCK

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - IF Confocal analysis of C6 cells using alpha Tubulin antibody (NB100-690, 1:50). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).

Immunocytochemistry/Immunofluorescence: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of embryonic fibroblasts in the anaphase portion of mitosis.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of paraffin embedded colon sections.

Immunohistochemistry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of small intestine tissue fixed with formalin and paraffin embedded showing cytoplasmic and cytoskeletal staining of glandular cells.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse skeletal muscle using alpha Tubulin Antibody (DM1A) at 1:100 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a strong cytoplasmic signal in the muscle cells with cytoplasmic-nuclear signal in the endothelial cells.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse lung using alpha Tubulin Antibody (DM1A) at 1:100 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated chunks of cytoplasmic signal in the alveolar and bronchiolar epithelial cells.

Immunohistochemistry-Paraffin: alpha Tubulin Antibody (DM1A) [NB100-690] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse heart using alpha Tubulin Antibody (DM1A) at 1:100 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a strong and specific cytoplasmic signal in the muscle cells.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of PE conjugate of NB100-690. An intracellular stain was performed on RAW 246.7 cells with Alpha Tubulin antibody (DM1A) NB100-690PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized wi

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - Analysis of PE conjugate of NB100-690. An intracellular stain was performed on SH-SY5Y cells with Alpha Tubulin antibody (DM1A) NB100-690PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized with

Flow (Intracellular): alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on U-937 cells with alpha Tubulin Antibody (DM1A) NB100-690AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647

Flow (Intracellular): alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin Antibody (DM1A) NB100-690AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488. Image from the Alexa Fluor 488 version of this antibody.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin (DM1A) Antibody NB100-690G (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 488.

Flow Cytometry: alpha Tubulin Antibody (DM1A) [NB100-690] - An intracellular stain was performed on HeLa cells with alpha Tubulin [DM1A] Antibody NB100-690AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunomicroscopy: alpha Tubulin Antibody (DM1A) [NB100-690] - Staining of the marine parasite Cryptocaryon irritans mouth. Large bundles of microtubules form a cytophyrigeal basket.

• Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Fu, Gp, Gt, Ha, Pa, Pm, Pm, Rb, Xp, Ye
• Applications: WB, Simple Western, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IM, IP, CyTOF-ready
• Clone: DM1A
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

alpha Tubulin Antibody (DM1A) Summary

• Description: As the TUBA1A gene is conserved evolutionarily and is ubiquitously expressed in most eukaryotic cell lines, the Alpha tubulin antibody has been shown to be an attractive and effective choice for a loading control, detecting at approximately 50-55 kDa. Quantitative western blotting requires a loading control in order to account and adjust for the differences in the loading of samples across wells.
• Immunogen: This alpha Tubulin Antibody (DM1A) was developed against native chicken brain microtubules.
• Marker: Microtubule Marker
• Specificity: This alpha Tubulin Antibody (DM1A) does not cross-react with beta Tubulin.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TUBA1A
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:5000
  • Simple Western 1:50
  • Flow Cytometry 1 ug per million cells
  • Immunocytochemistry/Immunofluorescence 1:1000
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Frozen 1:100-1:500
  • Immunohistochemistry-Paraffin 1:100-1:500
  • Immunomicroscopy
  • Immunoprecipitation 1:50-1:100
  • CyTOF-ready
  • Flow (Intracellular)
• Application Notes: This alpha Tubulin Antibody (DM1A) is useful as a loading control for Western blot as well as Immunoprecipitation, Immunohistochemistry on paraffin-embedded and frozen sections, Immunocytochemistry/Immunofluorescence and Flow Cytometry. The DM1A alpha tubulin antibody is ideal for use as a Western blot loading control, where a band can be seen around 50-55 kDa and as a cytoskeletal marker in ICC. For IHC-Paraffin, antigen retrieval is not essential, but may optimize staining.
  
  In Simple Western only 10 - 15 ul of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  
  This antibody is CyTOF ready.
• Theoretical MW: 50 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Tubulin alpha-1B Overexpression Lysate
  Human Lung Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

Human, mouse, rat, porcine, bovine, canine, chicken, guinea pig, Chinese hamster, monkey, avian, rabbit, Drosophila fungi and Xenopus. Yeast reactivity reported in scientific literature (PMID: 25126732). Goat reactivity reported in scientific literature (PMID:31805146). Will likely react with all mammals.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for alpha Tubulin Antibody (DM1A)

• alpha tubulin loading control
• alpha tubulin monoclonal
• alpha Tubulin
• Alpha-Tubulin 3
• B-ALPHA-1
• DM1A anti-tubulin
• dm1a monoclonal
• DM1a tubulin
• DM1A
• FLJ25113
• hum-a-tub1
• hum-a-tub2
• LIS3
• TUBA1 monoclonal
• TUBA1A
• TUBA3
• tubulin alpha-1A chain
• Tubulin alpha-3 chain
• Tubulin B-Alpha-1
• tubulin monoclonal
• tubulin, alpha 1a
• tubulin, alpha 3
• tubulin, alpha, brain-specific

Background

Tubulin is the major building block of microtubules. Microtubules function as structural and mobile elements in mitosis, intracellular transport, flagellar movement and in the cytoskeleton. The tubulin superfamily is composed of six distinct families. Genes from the alpha, beta and gamma tubulin families are found in all eukaryotes. Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulins. Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter with a theoretical molecular weight of 50 kDa.

Tyrosine ligase adds a C-terminal tyrosine to monomeric alpha tubulin. Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated alpha tubulin is referred to as Glu-tubulin. Another post-translational modification of detyrosinated alpha tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle.

Like GAPDH and beta-actin, alpha/beta tubulin is often used as a loading control in immunoblot applications (1). Alpha/beta tubulin is also good for counterstaining microtubules in immunofluorescence (2).

References

1. Hannen, R., Selmansberger, M., Hauswald, M., Pagenstecher, A., Nist, A., Stiewe, T.,... Bartsch, J. W. (2019). Comparative Transcriptomic Analysis of Temozolomide Resistant Primary GBM Stem-Like Cells and Recurrent GBM Identifies Up-Regulation of the Carbonic Anhydrase CA2 Gene as Resistance Factor. Cancers (Basel), 11(7). doi:10.3390/cancers11070921

2. Nel, M., Joubert, A. M., Dohle, W., Potter, B. V., & Theron, A. E. (2018). Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines. Drug Des Devel Ther, 12, 1881-1904. doi:10.2147/dddt.S152718

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for alpha Tubulin Antibody (NB100-690)(73)

Publications using NB100-690

• Kaid C, Madi RADS, Astray R et al. Safety, Tumor Reduction, and Clinical Impact of Zika Virus Injection in Dogs with Advanced-Stage Brain Tumors Mol. Ther. Mar 10 2020 [PMID: 32220305] (IHC-P, Canine)
• Jaworski Jp, Urrutia M, Dascal E et Al. C-C motif chemokine receptor 2 as a novel intermediate in the ovulatory cascade Mol. Hum. Reprod. Mar 11 2020 [PMID: 32159806]
• TRPM7 contributes to progressive nephropathy Sci Rep Feb 11 2020 [PMID: 32047249] (WB, Mouse)
• Chou CW, Lin CH, Hsiao TH et al. Therapeutic effects of statins against lung adenocarcinoma via p53 mutant-mediated apoptosis Sci Rep Dec 31 2019 [PMID: 31892709] (WB, Human)
• Yamada Y, Liao GR, Tseng CY, et al. Establishment and characterization of transformed goat primary cells by expression of simian virus 40 large T antigen for orf virus propagations PLoS ONE Dec 5 2019 [PMID: 31805146] (WB, Human, Primate, Goat)
• Doyle TB, Muntean BS, Ejendal KF et al. Identification of Novel Adenylyl Cyclase 5 (AC5) Signaling Networks in D1 and D2 Medium Spiny Neurons using Bimolecular Fluorescence Complementation Screening Cells. Nov 19 2019 [PMID: 31752385] (WB, Human)
• Murray JR, de la Vega L, Hayes JD et al. Induction of the Antioxidant Response by the Transcription Factor NRF2 Increases Bioactivation of the Mutagenic Air Pollutant 3-Nitrobenzanthrone in Human Lung Cells Chem. Res. Toxicol. Nov 20 2019 [PMID: 31746589]
• Seale LA, Khadka VS, Menor M et al. Combined Omics Reveals That Disruption of the Selenocysteine Lyase Gene Affects Amino Acid Pathways in Mice Nutrients 2019 Oct 26 [PMID: 31717805]
• Lee S, Eyer J, Letournel F et al. Neurofilaments form flexible bundles during neuritogenesis in culture and in mature axons in situ J. Neurosci. Res. Jul 15 2019 [PMID: 31304612] (ICC/IF, Mouse, Chicken, Canine)
• Tajima K, Matsuda S, Yae T et al. SETD1A protects from senescence through regulation of the mitotic gene expression program Nat Commun Jun 28 2019 [PMID: 31253781] (ICC/IF, Human)
• Lutz MM th, Worth MP, Hinchman MM et al. Mammalian orthoreovirus Infection is Enhanced in Cells Pre-Treated with Sodium Arsenite Viruses. 2019 Jun 17 [PMID: 31216693] (WB, Monkey)
• Doyle TB, Hayes MP, Chen DH et al. Functional characterization of AC5 gain-of-function variants: Impact on the molecular basis of ADCY5-related dyskinesia Biochem. Pharmacol. Feb 14 2019 [PMID: 30772269] (WB, Human)
• Miller, CN;Samuels, JS;Azhar, Y;Parmar, A;Shashidharamurthy, R;Rayalam, S; Guggulsterone Activates Adipocyte Beiging through Direct Effects on 3T3-L1 Adipocytes and Indirect Effects Mediated through RAW264.7 Macrophages Medicines (Basel) Jan 31 2019 [PMID: 30709026] (WB, Mouse)
• Kandel MB, Yamamoto S, Midorikawa R et al. N-glycosylation of the AMPA-type glutamate receptor regulates cell surface expression and tetramer formation affecting channel function J. Neurochem. Aug 9 2018 [PMID: 30092607] (WB, Human)
• Nel M, Joubert AM, Dohle W et al. Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines. Drug Des Devel Ther. Jun 25 2018 [PMID: 29983544] (ICC/IF, Human)
• Lin CY, Yang ST, Shen SC et al. Serum amyloid A1 in combination with integrin aVB3 increases glioblastoma cells mobility and progression Mol Oncol May 1 2018 [PMID: 29603594] (WB, Human)
• Tomoshige S, Kobayashi Y, Hosoba K et al. Cytoskeleton-related regulation of primary cilia shortening mediated by melanin-concentrating hormone receptor 1 Gen. Comp. Endocrinol. 2017 Aug 22 [PMID: 28842217] (WB, Human)
• Zhang M, Yu Q, Liu Z et al. UBX domain-containing proteins are involved in lipid homeostasis and stress responses in Pichia pastoris Int. J. Biochem. Cell Biol. 2017 Aug 12 [PMID: 28807601] (WB, Yeast)
• Vent-Schmidt RYJ, Wen RH, Zong Z et al. Opposing Effects of Valproic Acid Treatment Mediated by Histone Deacetylase Inhibitor Activity in Four Transgenic X. laevis Models of Retinitis Pigmentosa J Neurosci. 2017 Jan 24 [PMID: 28490005] (WB, Xenopus)
• Chen PC, Hsieh MH, Wen-ShuoKuo P et al. Water-soluble chitosan inhibits nerve growth factor and attenuates allergic inflammation in mite allergen-induced allergic rhinitis. J. Allergy Clin. Immunol. Apr 12 2017 [PMID: 28412394] (WB)
• Sidoli M, Musner N, Silvestri N et al. Ablation of Perk in Schwann Cells Improves Myelination in the S63del Charcot-Marie-Tooth 1B Mouse. J. Neurosci. Nov 2 2016 [PMID: 27807175] (WB, Mouse)
• Rueli RH, Torres DJ, Dewing AS et al. Selenoprotein S Reduces Endoplasmic Reticulum Stress-Induced Phosphorylation of Tau: Potential Role in Selenate Mitigation of Tau Pathology. J. Alzheimers Dis. Oct 11 2016 [PMID: 27802219]
• Chang CF, Hsu LS, Weng CY et al. N-Glycosylation of Human R-Spondin 1 Is Required for Efficient Secretion and Stability but Not for Its Heparin Binding Ability. Int J Mol Sci Jun 18 2016 [PMID: 27314333] (WB,)
• Macedo CG, Napimoga MH, Rocha-Neto LM et al. The role of endogenous opioid peptides in the antinociceptive effect of 15-deoxy(delta12,14)-prostaglandin J2 in the temporomandibular joint. Prostaglandins Leukot. Essent. Fatty Acids [PMID: 27255640] (WB)
• Park HS, Jang JE, Ko MS et al. Statins Increase Mitochondrial and Peroxisomal Fatty Acid Oxidation in the Liver and Prevent Non-Alcoholic Steatohepatitis in Mice. Diabetes Metab J. 2016 Apr 21 [PMID: 27098507] (WB, Mouse)
• Tu DG, Yu Y, Lee CH et al. Hinokitiol inhibits vasculogenic mimicry activity of breast cancer stem/progenitor cells through proteasome-mediated degradation of epidermal growth factor receptor. Oncol Lett 2016 Apr [PMID: 27073579] (WB)
• Wang TK, Lin YM, Lo CM et al. Oncogenic roles of carbonic anhydrase 8 in human osteosarcoma cells. Tumour Biol. 2015 Dec 28 [PMID: 26711783] (WB, Human)
  
  Details:
  alpha Tubulin antibody (clone DM1A) was used for WB assays in experiments involving human osteosarcoma cell lines MG-63, U-2 OS, HOS and 143B and human embryonic kidney cell line HEK293.
• Farnsworth NL, Walter RL, Hemmati A et al. Low Level Pro-Inflammatory Cytokines Decrease Connexin36 Gap Junction Coupling in Mouse and Human Islets through Nitric Oxide Mediated Protein Kinase Cdelta. J. Biol. Chem. 2015 Dec 14 [PMID: 26668311] (WB, Mouse)
  
  Details:
  alpha Tubulin antibody (clone DM1A) used for WB on mouse islets at a dilution of 1:5,000. alpha Tubulin antibody was used as a loading control (Fig. 6D).
• Pitts MW, Kremer PM, Hashimoto AC et al. Competition between the Brain and Testes under Selenium-Compromised Conditions: Insight into Sex Differences in Selenium Metabolism and Risk of Neurodevelopmental Disease. J. Neurosci. 2015 Nov 18 [PMID: 26586820] (WB, Mouse)
  
  Details:
  Alpha Tubulin antibody used for WB as a loading control at a dilution of 1:5000 on mouse brainstem and whole-brain samples (Figure 6A).
• Alleaume-Butaux A, Nicot S, Pietri M et al. Double-Edge Sword of Sustained ROCK Activation in Prion Diseases through Neuritogenesis Defects and Prion Accumulation. PLoS Pathog. 2015 Aug 01 [PMID: 26241960] (ICC/IF, WB, Mouse)
• Seale LA, Gilman CL, Hashimoto AC et al. Diet-induced obesity in the selenocysteine lyase knockout mouse Antioxid. Redox Signal. 2015 Jul 20 [PMID: 26192035]
• Zhang W, Ogando DG, Bonanno JA, Obukhov AG. Human SLC4A11 is a Novel NH3: H+ Co-transporter J. Biol. Chem. 2015 May 27 [PMID: 26018076] (WB, Human)
• Bachman MJ, Foltz K, Lynch JM et al. Using cytochrome P4501A1 expression in liver and blubber to understand effects of persistent organic pollutant exposure in stranded pacific island cetaceans Environ. Toxicol. Chem. 2015 Apr 18 [PMID: 25892359] (WB)
• Sulaiman MK, Chu Z, Blanco VM et al. SapC-DOPS nanovesicles induce Smac- and Bax-dependent apoptosis through mitochondrial activation in neuroblastomas Mol. Cancer 2015 Apr 19 [PMID: 25889084] (WB, Human)
• Stander XX, Stander BA, Joubert AM. Synergistic Anticancer Potential of Dichloroacetate and Estradiol Analogue Exerting their Effect via ROS-JNK-Bcl-2-Mediated Signalling Pathways Cell. Physiol. Biochem. 2015 Mar 31 [PMID: 25791820] (WB, Human)
• Munoz-Saez E, de Munck Garcia E, Arahuetes Portero RM et al. Neuroprotective role of sphingosine-1-phosphate in L-BMAA treated neuroblastoma cells (SH-SY5Y). Neurosci. Lett. 2015 Mar 10 [PMID: 25769802] (WB, Human)
  
  Details:
  Alpha-Tubulin antibody (DM1A) used for WB as a loading control on total extract and nucleus from SH-SY5Y cells (Fig. 3 & 4).
• Eriksen AB, Torgersen ML, Holm KL et al. Retinoic acid-induced IgG production in TLR-activated human primary B cells involves ULK1-mediated autophagy Autophagy 2015 Mar 06 [PMID: 25749095] (WB, Human)
• Park SJ, Jeong J, Park YU et al. Disrupted-in-schizophrenia-1 (DISC1) Regulates Endoplasmic Reticulum Calcium Dynamics Sci Rep 2015 Mar 03 [PMID: 25732993] (WB, Human)
• Hsuan SW, Chyau CC, Hung HY et al. The induction of apoptosis and autophagy by Wasabia japonica extract in colon cancer Eur J Nutr 2015 Feb 27 [PMID: 25720497] (WB, Human)
• Xu N, Dong YJ, Yu QL et al. Convergent Regulation of Candida albicans Aft2 and Czf1 in Invasive and Opaque Filamentation J. Cell. Biochem. 2015 Feb 26 [PMID: 25716417] (WB, Fungi)
  
  Details:
  Alpha tubulin antibody used for WB in experiments involving several stains of Candida albicans.
• Yu Q, Jia C, Dong Y et al. Candida albicans autophagy, no longer a bystander: Its role in tolerance to ER stress-related antifungal drugs. Fungal Genet. Biol. 2015 Feb 21 [PMID: 25711686] (WB, Fungi)
  
  Details:
  Alpha-Tubulin antibody (DM1A) used for WB as a loading control on fungal cells that were untreated or treated with tunicamycin (Fig. 8A).
• Rucker H, Amslinger S. Identification of heme oxygenase-1 stimulators by a convenient ELISA-based bilirubin quantification assay. Free Radical Biology and Medicine. 2014 Oct 23 [PMID: 25462643] (WB, Mouse)
  
  Details:
  alpha Tubulin antibody used as a loading control for WB application (1:500 dilution, ECL detection, sharp band at 57kDa position) in experiments involving stimulation of murine macrophage RAW264.7 cells.
• Yu Q, Zhang B, Yang B et al. Interaction between the Vacuole, the Mitochondria and Oxidative Stress Response Is Governed by the Transient Receptor Potential Channel in Candida albicans. Free Radic. Biol. Med. 2014 Oct 09 [PMID: 25308698] (WB)
• Levine B. Autophagy is required for G1/G0 quiescence in response to nitrogen starvation in Saccharomyces cerevisiae. Autophagy. 2014 Aug 12 [PMID: 25126732] (ICC/IF, Yeast)
  
  Details:
  alpha Tubulin antibody used for ICC-IF on yeast strains - VPS30 delta/ATG6 delta, ATG7 delta, PEP4 delta subjected or not to starvation with nitrogen starvation medium/SD-N for 4 hours (Figure 4A).
• Doria ML,Ribeiro AS,Wang J et al. Fatty acid and phospholipid biosynthetic pathways are regulated throughout mammary epithelial cell differentiation and correlate to breast cancer survival FASEB J. 2014 Jun 26 [PMID: 24970396] (WB, Mouse)
  
  Details:
  alpha Tubulin antibody used as loading control in WB for lysates of differentiating mammary epithelial cell line HC11 (Figure 3B, Figure 4C)
• Stoeckelhuber M, Olzowy B, Ihler F et al. Immunolocalization of antimicrobial and cytoskeletal components in the serous glands of human sinonasal mucosa. Histol. Histopathol. 2014 Apr 15 [PMID: 24737387] (IHC-P, Human)
• Weng CM, Yu CC, Kuo ML, Lin CH. Endothelin-1 induces connective tissue growth factor expression in human lung fibroblasts by ETAR-dependent JNK/AP-1 pathway. Biochem. Pharmacol. 2014 Feb 21 [PMID: 24486572] (WB, Human)
• Hsieh YT, Chou MM, Chen HC, Tseng JJ. IMP1 promotes choriocarcinoma cell migration and invasion through the novel effectors RSK2 and PPME1. Gynecol Oncol. 2013 Oct [PMID: 23911878]
• Barayuga SM, Pang X, Andres MA et al. Methamphetamine decreases levels of glutathione peroxidases 1 and 4 in SH-SY5Y neuronal cells: Protective effects of selenium. Neurotoxicology 2013 May 27 [PMID: 23721877] (WB, Human)
• Cheng X, Xu N, Yu Q et al. Novel insight into the expression and function of the multicopper oxidases in Candida albicans. Microbiology 2013 Apr 11 [PMID: 23579686] (WB, Fungi)
• Chandley MJ, Szebeni K, Szebeni A et al. Gene expression deficits in pontine locus coeruleus astrocytes in men with major depressive disorder. J Psychiatry Neurosci 2013 Jul [PMID: 23415275] (WB, Human)
• Kim KW, Lee JM, Jeon YS et al. Vascular disrupting effect of CKD-516: preclinical study using DCE-MRI Invest New Drugs 2013 Jan 9 [PMID: 23299389] (ICC/IF, IHC-Fr, Human, Rabbit)
• Cavallo F, Graziani G, Antinozzi C et al. Reduced proficiency in homologous recombination underlies the high sensitivity of embryonal carcinoma testicular germ cell tumors to Cisplatin and poly (adp-ribose) polymerase inhibition PLoS One 2012 [PMID: 23251575] (WB, Human, Mouse)
• Gobeil PA, Leib DA. Herpes Simplex Virus gamma34.5 Interferes with Autophagosome Maturation and Antigen Presentation in Dendritic Cells MBio 2012 Oct 16 [PMID: 23073763] (WB, Mouse)
• Seale LA, Hashimoto AC, Kurokawa S et al. Disruption of the selenocysteine lyase-mediated selenium recycling pathway leads to metabolic syndrome in mice Mol Cell Biol 2012 Oct [PMID: 22890841] (WB, Mouse)
• Gossett DR, Tse HT, Lee SA, Ying Y, Lindgren AG, Yang OO, Rao J, Clark AT, Di Carlo D. Hydrodynamic stretching of single cells for large population mechanical phenotyping. Proc Natl Acad Sci U S A. 2012 Apr 30. [PMID: 22547795] (ICC/IF, Human)
• Raman AV, Pitts MW, Seyedali A, Hashimoto AC, Seale LA, Bellinger FP, Berry MJ. Absence of selenoprotein P but not selenocysteine lyase results in severe neurological dysfunction. Genes Brain Behav. 2012 Apr 6. [PMID: 22487427] (WB, Mouse)
• Burmeister D, Aboushwareb T, D'Agostino R Jr et al. Impact of partial urethral obstruction on bladder function: time-dependent changes and functional correlates of altered expression of Ca2+ signaling regulators Am J Physiol Renal Physiol 2012 Jun [PMID: 22442207] (WB, Rat)
• Ko WC, Chen BC, Hsu MJ et al. Thrombin induced connective tissue growth factor expression in rat vascular smooth muscle cells via the PAR-1/JNK/AP-1 pathway. Acta Pharmacol Sin. 2012 Jan [PMID: 22212430] (WB, Rat)
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Reviews for alpha Tubulin Antibody (NB100-690) (14)

Average Rating: 4.6

(Based on 14 reviews)

We have 14 reviews tested in 4 species: Human, Mouse, Human and Mouse, Other.

reviewed by: Anonymous WB Human 05/18/2020

Summary

• Application: Western Blot
• Sample Tested: Breast cancer cells
• Species: Human
• Lot: A-4

reviewed by: Anonymous WB Human 10/22/2018

Summary

• Application: Western Blot
• Sample Tested: Human breast cancer cell lines
• Species: Human
• Lot: A-4

reviewed by: Jun Hamazaki WB Human and Mouse 06/29/2017

Summary

• Application: Western Blot
• Sample Tested: 293T cell lysate,nih3t3 cell lysate
• Species: Human and Mouse
• Lot: e-3

reviewed by: Emily Ledgerwood WB Other 07/15/2016

Summary

• Application: Western Blot
• Sample Tested: African Green Monkey Kidney (CV-1) whole cell lysate
• Species: Other
• Lot: E-1

Blocking

• Blocking Details: 1 hour in 5% milk/TBST

Primary Anitbody

• Dilution Ratio: alpha tubulin NB100-690 1:5000 in 5% milk/TBST overnight at 4 degrees. This is a membrane that was stripped and reprobed.

Secondary Antibody

• Secondary Description: Donkey anti-mouse HRP
• Secondary Manufacturer Cat#: 715-035-150
• Secondary Concentration: 1:10000

Details

• Detection Notes: Exposed on a LiCor C-Digit using Advansta WesternBright Quantum

Comments

• Comments: This is a fantastic antibody for alpha-tubulin that I have been using for years. For this review, I opted to submit an image of a PVDF membrane that was originally probed for a different primary, stripped using LiCor's Western Blot Stripping Buffer according to manufacturer's instructions and then reprobed with this alpha-tubulin antibody. To top it off, I am imaging on a C-digit which is the cheapest imager on the market. This antibody is extremely reliable and well worth the money.

reviewed by: Anony- mous WB Human 06/03/2014

Summary

• Application: Western Blot
• Species: Human
• Special Applications: Western blot analysis of extracts from HeLa, COS and C6 cells using alpha Tubulin antibody (NB100-690, 1:1000).
• File: View PDF

reviewed by: Anony- mous ICC Human 06/03/2014

Summary

• Application: Immunocytochemistry
• Species: Human
• Special Applications: IF Confocal analysis of C6 cells using alpha Tubulin antibody (NB100-690, 1:50). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).
• File: View PDF

reviewed by: Samantha Garcia ICC Human 11/06/2013

Summary

• Application: Immunocytochemistry
• Sample Tested: MDA-MB-231 cells
• Species: Human
• Lot: A2

Blocking

• Blocking Details: PBS with 2% BSA

Primary Anitbody

• Dilution Ratio: 1:2000 in blocking buffer

Details

• Fixation Details: MDA-MB-231 cells were fixed in 4% PFA. Glutaraldehyde fixation also worked.

reviewed by: Anonymous WB Mouse 01/06/2012

Summary

• Application: Western Blot
• Sample Tested: mouse lung tissue
• Species: Mouse

Blocking

• Blocking Details: Blocking Buffer: 100ug FBS

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: Mouse, Secondary Ab Dilution Ratio: 1:10000

Details

• Detection Notes: Detection Method: ECL, Exposure Time: 1 minute, Specific Bands: 51 kDa, Non-Specific Bands: 50-55 kDa

reviewed by: Anonymous WB Mouse 12/05/2011

Summary

• Application: Western Blot
• Sample Tested: Cell lysate
• Species: Mouse
• Comments: The signal is strong and clear.

Blocking

• Blocking Details: Blocking Buffer: 5% BSA

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: 2 hours

Secondary Antibody

• Secondary Description: Secondary Ab: mouse HRP

Details

• Detection Notes: Detection Method: ECL with X-ray film

Comments

• Comments: The signal is strong and clear.

reviewed by: Anonymous WB Human 11/23/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human
• Comments: Worked well

Blocking

• Blocking Details: Blocking Time: 1 hour

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: mouse

Details

• Detection Notes: Detection Method: HRP

Comments

• Comments: Worked well

reviewed by: Anonymous WB Human 11/23/2011

Summary

• Application: Western Blot
• Sample Tested: Human cell
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: BSA

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: anti-mouse

Details

• Detection Notes: Detection Method: HRP

reviewed by: Anonymous WB Human 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 5% non-fat milk in TBST

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:100, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: anti-rabbit

Details

• Detection Notes: Detection Method: Western Lighting plus ECL

reviewed by: Anonymous WB Human 11/18/2011

Summary

• Application: Western Blot
• Sample Tested: Human cell
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: 1 hour, Primary Ab Incubation Temp: room temperature

Secondary Antibody

• Secondary Description: Secondary Ab: HRP

Details

• Detection Notes: Detection Method: chemiluminescence system

reviewed by: Anonymous WB Human 10/31/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human
• Comments: Great performance! Clear band in signal

Blocking

• Blocking Details: Blocking Buffer: 5% Skim Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:10000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: HRP conjugated anti-mouse

Details

• Detection Notes: Detection Method: Chemiluminescence

Comments

• Comments: Great performance! Clear band in signal

FAQs for alpha Tubulin Antibody (NB100-690). (Showing 1 - 3 of 3 FAQs).

1. I am interested in buying an actin or tubulin antibody for Western blot loading control. We would like to use it on rat heart samples. Please recommend us one of your actin and/or tubulin products, and add it to the NOX2 quotation.
   • NB100-690 is for Alpha Tubulin and has been validated for use in WB and Rat. This will be added to the quote along with our Actin Antibody.
2. I am working on a dissection fixation protocol for a non model organism, and I am using DM1A alpha tubulin as a control.  I do not see any sort of Astral projections; does this convince you that it is a working stain ?
   • The staining looks a little off to my eye. What were your fixation conditions? Tubulin immunostaining can be finicky and requires particular fixation conditions for optimal labeling. Depending on your other antigenic sites, it's ideal to use either mixed aldehydes (PFA and Glutaraldehyde) with simultaneous extraction or methanol. PFA alone gives an undesirable hazy staining pattern.Some laboratories also recommend adding a calcium chelator into the fixation buffer as calcium induces depolymerization of microtubules. Lastly, be sure to run negative controls with your staining.
3. We would like to stain cilia with an acetylated alpha tubulin antibody in our cells, but I am unsure if this antibody will be able to conclusively differentiate cilia from other structures such as spindle pole bodies. Does anyone know what acetylated alpha tubulin antibodies might bind to apart from cilia?
   • Acetylated alpha tubulin is found in relatively stable microtubules. It is best practice to use this marker together with a centrosome/centriole marker, which will stain the basal bodies at the base of the cilium. After that, it is relatively straightforward to identify the acetylated alpha tubulin signal that corresponds to the cilium.

Bioinformatics

• Gene Symbol: TUBA1A
• Entrez:
  • 7846(Human)