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RabMAb

Anti-alpha Tubulin 抗体 [EP1332Y] - Microtubule Marker (ab52866)

製品の概要

  • 製品名

    Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker
    alpha Tubulin 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: ICC/IF, IHC - Wholemount, WB, Flow Cyt, IHC-P, IHC-Frmore details
  • 種交差性

    交差種: Mouse, Rat, Human, Pig, Drosophila melanogaster
  • 免疫原

    Synthetic peptide within Human alpha Tubulin aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P68366

  • ポジティブ・コントロール

    • WB- HeLa, HEK-293, HepG2, Caco2, NIH/3T3, PC-12, RAW 264.7, PC-12, C6 Jurkat and HEK-293T whole cell lysates; human fetal kidney lysate; Mouse and rat brain lysate; Pig skeletal muscle lysates; IHC-P: Pig kidney tissue; rat kidney tissue; mouse kidney tissue; human breast cancer and stomach tissue; IHC-Fr: Rat kidney tubule tissue; Flow cyt: HepG2 cells; ICC/IF: HUVEC, HeLa and 293 cells.
  • 特記事項

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab52866 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF 1/250 - 1/500.
IHC - Wholemount Use at an assay dependent concentration.
WB 1/1000 - 1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
Flow Cyt 1/20 - 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration. PubMed: 21933451

ターゲット情報

  • 機能

    Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
  • 配列類似性

    Belongs to the tubulin family.
  • 翻訳後修飾

    Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
    Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
  • 細胞内局在

    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 参照データベース

  • 別名

    • Alpha-tubulin 1 antibody
    • ALS22 antibody
    • B ALPHA 1 antibody
    • bA408E5.3 antibody
    • H2 ALPHA antibody
    • Hum a tub1 antibody
    • Hum a tub2 antibody
    • LIS3 antibody
    • MGC171407 antibody
    • MGC55332 antibody
    • TBA4A_HUMAN antibody
    • Testis-specific alpha-tubulin antibody
    • TUBA1 antibody
    • TUBA1A antibody
    • tuba1l antibody
    • Tuba4a antibody
    • Tubulin alpha 1 chain antibody
    • Tubulin alpha antibody
    • Tubulin alpha-1 chain antibody
    • tubulin alpha-1B chain antibody
    • Tubulin alpha-4A chain antibody
    • Tubulin H2-alpha antibody
    • Tubulin, alpha 1 (testis specific) antibody
    • tubulin, alpha 1, like antibody
    • Tubulin, alpha 4a antibody
    • Tubulin, alpha, testis-specific antibody
    • Tubulin, alpha-1 antibody
    see all

画像

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.

     

  • All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/10000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates
    Lane 2 : PC-12 (Rat adrenal gland heochromocytoma) whole cell lysates
    Lane 3 : NIH/3T3( Mouse embryonic fibroblast)whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling alpha Tubulin with ab52866 at   followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human stomach.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol  permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling alpha Tubulin with ab52866 at 1/2000 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.  

  • All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/5000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : C6 (Rat glial tumor cell line) whole cell lysates
    Lane 3 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysates
    Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 50 kDa

  • Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/100 dilution.

  • Flow cytometry analysis of 2% paraformaldehyde fixed HepG2 (human liver hepatocellular carcinoma cell line) cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

  • Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of rat kidney tubule tissue, staining alpha Tubulin with ab52866.
  • AJAP1 co-localizes with microtubules in HUVECs

    The association of AJAP1 with microtubules in HUVECs is lost upon microtubule destruction. Treatment with 12.5 µM nocodazole for 24 h shows destruction of the microtubule network and loss of AJAP1 tubular localization. For a negative control, HUVECs are treated with DMSO for 24 h. Cell nuclei were counterstained with DAPI (cyan). Microscope: Zeiss LSM 780; objective lens: 63×/1.40 oil; scale bar: 25 µm.

    Incubated overnight at 4°C with ab52866.

    (From Figure 3E of Hotte et al)

  • All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/20000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 50 kDa

  • Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/50000 dilution + Rat brain lysates at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 50 kDa

  • ab52866 staining alpha Tubulin in 293 Human embryonic kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 2 hours at 23°C. Samples were incubated with primary antibody (1/200 in 0.5% saponin) for 2 hours at 23°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.

    See Abreview

  • Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

  • Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

     

  • Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/50000 dilution + Human fetal kidney lysates at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 50 kDa

  • All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52866 overnight at 4°C. Antibody binding was detected using Anti-Rabbit Alexa Fluor® 790 (ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/5000 dilution + Pig skeletal muscle lysates at 20 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 50 kDa

参考文献

This product has been referenced in:

  • Lee M  et al. In-depth study of lin-28 suggests selectively conserved let-7 independent mechanism in Drosophila. Gene 687:64-72 (2019). Read more (PubMed: 30415010) »
  • Takanezawa Y  et al. An autophagy deficiency promotes methylmercury-induced multinuclear cell formation. Biochem Biophys Res Commun 511:460-467 (2019). Read more (PubMed: 30797556) »
See all 84 Publications for this product

レビューと Q&A

1-10 of 26 Abreviews or Q&A

Application
Western blot
Loading amount
62.5 µg
Gel Running Conditions
Reduced Denaturing (12.5% acrylamide)
Sample
Pig Tissue lysate - other (Muscle sarcoplasmic extract)
Specification
Muscle sarcoplasmic extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Shannon Cruzen

Verified customer

投稿 Feb 05 2014

Application
IHC - Wholemount
Sample
Fruit fly (Drosophila melanogaster) Tissue (Male gonad)
Specification
Male gonad

Abcam user community

Verified customer

投稿 Jul 05 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No

Abcam user community

Verified customer

投稿 Feb 28 2012

Application
Western blot
Sample
Rat Cell lysate - whole cell (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (10 %)
Blocking step
Odyssey blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C

Abcam user community

Verified customer

投稿 Jun 17 2009

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (MC3T3)
Permeabilization
Yes - NP40
Specification
MC3T3
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Aug 16 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MC3T3)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
MC3T3
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

投稿 Aug 14 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Vascular smooth muscle cells)
Permeabilization
Yes - NP40
Specification
Vascular smooth muscle cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Oct 22 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Elephant Cell (Sperm)
Permeabilization
Yes - BSA-T
Specification
Sperm
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Glutaraldehyde

Abcam user community

Verified customer

投稿 May 31 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (osteosarcoma)
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Loading amount
40 µg
Treatment
Plasmid transfection using HD fuGENE
Specification
osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Abcam user community

Verified customer

投稿 Jun 27 2017

Application
Western blot
Sample
Human Cell lysate - other (cytoplasmic anf nuclear fraction from skin keratin)
Gel Running Conditions
Reduced Denaturing (any Kd BioRad)
Loading amount
20 µg
Specification
cytoplasmic anf nuclear fraction from skin keratin
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Julien Coutier

Verified customer

投稿 Feb 07 2017

1-10 of 26 Abreviews or Q&A

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