RabMAb

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

Overview

  • Product nameAnti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker
    See all alpha Tubulin primary antibodies
  • Description
    Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker
  • Tested applicationsICC/IF, IHC - Wholemount, WB, IP, Flow Cyt, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig, Fruit fly (Drosophila melanogaster)
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) (N terminal)

  • Positive control
    • This antibody gave a positive signal within WB in the following whole cell lysates: HeLa; HEK293; HepG2; Caco2; NIH3T3; PC12. HeLa cell lysate, human gastric carcinoma tissue or HeLa cells.
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A 40 μl trial size is available to purchase for this antibody.

    The Alexa Fluor® 488 variant is available, see ab185031.

    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.

Properties

Applications

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.

Application Abreviews Notes
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).
IP 1/50.
Flow Cyt 1/20 - 1/50.
IHC-P Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent concentration. PubMed: 21933451

Target

  • FunctionTubulin 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.
  • Sequence similaritiesBelongs to the tubulin family.
  • Post-translational
    modifications
    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.
  • Cellular localizationCytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha tubulin ubiquitous antibody
    • Alpha-tubulin 1 antibody
    • B ALPHA 1 antibody
    • bA408E5.3 antibody
    • H2 ALPHA antibody
    • Hum a tub1 antibody
    • Hum a tub2 antibody
    • LIS3 antibody
    • TBA4A_HUMAN antibody
    • Testis-specific alpha-tubulin antibody
    • TUBA1 antibody
    • TUBA1A antibody
    • Tuba4a antibody
    • Tubulin alpha 1 chain antibody
    • Tubulin alpha antibody
    • Tubulin alpha ubiquitous chain antibody
    • Tubulin alpha-1 chain antibody
    • Tubulin alpha-4A chain antibody
    • Tubulin H2-alpha antibody
    • Tubulin K alpha 1 antibody
    • Tubulin, alpha 1 (testis specific) antibody
    • Tubulin, alpha 4a antibody
    • Tubulin, alpha, testis-specific antibody
    • Tubulin, alpha-1 antibody
    see all

Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker images

  • alpha Tubulin was immunoprecipitated from 1mg of HeLa whole cell extract using ab52866 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab52866 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
    Lane 1 Hela whole cell extract, Lane 2 PBS instead of whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST. 

  • Flow cytometry analysis of 2% paraformaldehyde fixed HepG2 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).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Goat anti-rabbit IgG (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 ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab52866 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • 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. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) 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.

  • 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 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.

  • 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 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.

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

    Lane 1 : Mouse brain lysates
    Lane 2 : C6 whole cell lysates
    Lane 3 : Raw264.7 whole cell lysates
    Lane 4 : PC-12 whole cell lysates
    Lane 5 : NIH3T3 whole cell lysates

    Lysates/proteins at 10 µg per lane.

    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 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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 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 1/50000 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
  • 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
  • All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/20000 dilution

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HepG2 whole cell lysate
    Lane 3 : Jurkat whole cell lysate
    Lane 4 : 293T whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated 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

  • Immunohistochemical analysis of rat kidney tubule tissue, staining alpha Tubulin with ab52866.

References for Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)

This product has been referenced in:
  • Anchan D  et al. GPR30 activation decreases anxiety in the open field test but not in the elevated plus maze test in female mice. Brain Behav 4:51-9 (2014). WB ; Mouse . Read more (PubMed: 24653954) »
  • Bledau AS  et al. The H3K4 methyltransferase Setd1a is first required at the epiblast stage, whereas Setd1b becomes essential after gastrulation. Development 141:1022-35 (2014). Read more (PubMed: 24550110) »

See all 14 Publications for this product

Product Wall

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
Username

Dr. Shannon Cruzen

Verified customer

Submitted Feb 05 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application IHC - Wholemount
Sample Fruit fly (Drosophila melanogaster) Tissue (Male gonad)
Specification Male gonad
Username

Abcam user community

Verified customer

Submitted Jul 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Feb 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Jun 17 2009

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - other (vascular smooth muscle cell)
Specification vascular smooth muscle cell
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Oct 24 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (cultured cortical neurons at 20 DIV)
Specification cultured cortical neurons at 20 DIV
Fixative Methanol
Permeabilization Yes - 0,5%(w/v) saponin
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Feb 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Huh7 Human Hepatoma Cells)
Specification Huh7 Human Hepatoma Cells
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Saponin
Username

Abcam user community

Verified customer

Submitted Jan 06 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (cultured cortical neurons)
Loading amount 20 µg
Specification cultured cortical neurons
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Dec 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (293 human embryonic kidney)
Specification 293 human embryonic kidney
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Sep 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (293 human embryonic kidney)
Loading amount 25 µg
Specification 293 human embryonic kidney
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 31 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"