Overview

  • Product name
    Anti-alpha Tubulin antibody - Loading Control
    See all alpha Tubulin primary antibodies
  • Description
    Chicken polyclonal to alpha Tubulin - Loading Control
  • Host species
    Chicken
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Monkey
  • Immunogen

    Synthetic peptide corresponding to Human alpha Tubulin aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab23537)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; HepG2; Caco2; HCT116; NIH 3T3; PC12; ICC/IF: Caco-2 cells, NIH3T3 cells, SV40LT-SMC cells

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab89984 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 50 kDa).
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

  • Function
    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.
  • Sequence similarities
    Belongs 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 localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • All lanes : Anti-alpha Tubulin antibody - Loading Control (ab89984) at 1 µg/ml

    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 : HCT 116 (Human Colorectal Carcinoma) Whole Cell Lysate
    Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Chicken IgY H&L (Alexa Fluor® 790) (ab175787) 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 ab89984 overnight at 4°C. Antibody binding was detected using ab175787 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • ab89984 staining alpha Tubulin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab89984 at a working concentration of 1μg/ml and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-chicken AlexaFluor® 488 (ab150173) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
    This product also gave a positive signal in 100% methanol (5 min) fixed SV40 cells under the same testing conditions.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • All lanes : Anti-alpha Tubulin antibody - Loading Control (ab89984) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Chicken IgY - H&L (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 53 kDa why is the actual band size different from the predicted?
    Additional bands at: 125 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds
  • ab89984 staining alpha Tubulin in Caco-2 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab89984 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Chicken secondary (ab150173) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • ab89984 staining Tubulin in NIH3T3 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab89984 at 5μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ICC/IF image of ab89984 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab89984 at 5ug overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti- chicken IgY (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in HepG2 PFA fixed cell types at 5ug/ml.

References

This product has been referenced in:
  • Di Francesco L  et al. Visualization of human karyopherin beta-1/importin beta-1 interactions with protein partners in mitotic cells by co-immunoprecipitation and proximity ligation assays. Sci Rep 8:1850 (2018). IF . Read more (PubMed: 29382863) »
  • Shah M  et al. Profiling Subcellular Protein Phosphatase Responses to Coxsackievirus B3 Infection of Cardiomyocytes. Mol Cell Proteomics 16:S244-S262 (2017). Read more (PubMed: 28174228) »
See all 18 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (MDCK)
Permeabilization
Yes - 0.5% Triton X-100
Specification
MDCK
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 29 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cell lines: SH-SY5Y (neuronal, human) MLP29 (hepat)
Gel Running Conditions
Non-reduced Denaturing (4-12%)
Loading amount
20 µg
Specification
Cell lines: SH-SY5Y (neuronal, human) MLP29 (hepat
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 30 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - PBST for 30 minutes
Specification
HeLa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
4% formaldehyde in PBS/0.2% Triton X-100

Weiguo Zhang

Verified customer

Submitted Aug 25 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cell (Drosophila S2 cells)
Specification
Drosophila S2 cells
Fixative
Formaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Ms. Zuni Bassi

Verified customer

Submitted May 20 2011

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

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