Recombinant
RabMAb

Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560)

Overview

  • Product name
    Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control
    See all alpha Tubulin primary antibodies
  • Description
    Rabbit monoclonal [EPR13478(B)] to alpha Tubulin - Loading Control
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human alpha Tubulin aa 400 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: P68366

  • Positive control
    • HeLa, Jurkat, A431 and K562 cell lysates; Human kidney and uterus tissues; A431 and Jurkat cells.
  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab176560 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 1/2000 - 1/10000. Predicted molecular weight: 50 kDa.

For unpurified, use 1/1000 - 1/10000.

IHC-P 1/350. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified, use 1/100.

ICC/IF 1/350.

For unpurified, use 1/100.

Flow Cyt 1/60.

For unpurified, use 1/10.

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

IP 1/100.

For unpurified, use 1/30.

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

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human uterus tissue labeling alpha Tubulin with unpurified ab176560 at a 1/100 dilution.

  • Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560) at 1/7000 dilution (Purified) + Jurkat cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 50 kDa
    Observed band size: 50 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human kidney with purified ab176560 at a dilution of 1/350. A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was counter stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunofluorescent staining of A431 cells fixed in 4% PFA with purified ab176560 at a dilution of 1/350. An Alexa Fluor® 555 goat anti-rabbit was used as the secondary at a dilution of 1/500 and the sample was counter stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

  • All lanes : Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560) at 1/1000 dilution (Unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 50 kDa



    Secondary antibodies - anti-rabbit HRP (ab6721)

  • Immunofluorescent staining of A431 cells fixed in 4% PFA with unpurified ab176560 at a dilution of 1/100. An Alexa Fluor® 555 goat anti-rabbit was used as the secondary at a dilution of 1/500 and the sample was counter stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

  • Immunohistochemical staining of paraffin embedded human kidney with unpurified ab176560 at a dilution of 1/100. A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was counter stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Flow cytometry analysis of permeabilized HeLa cells labeling alpha Tubulin (pink) with purified ab176560 at a 1/70 dilution, or negative control rabbit IgG (green). The secondary antibody was FITC goat anti-rabbit.

  • Flow cytometry analysis of permeabilized Jurkat cells labeling alpha Tubulin (red) with unpurified ab176560 at a 1/10 dilution, or negative control rabbit IgG (green)

References

This product has been referenced in:
  • Mohamoud HS  et al. A missense mutation in TRAPPC6A leads to build-up of the protein, in patients with a neurodevelopmental syndrome and dysmorphic features. Sci Rep 8:2053 (2018). WB ; Human . Read more (PubMed: 29391579) »
  • Deng X  et al. Guanxin Danshen Formulation Protects against Myocardial Ischemia Reperfusion Injury-Induced Left Ventricular Remodeling by Upregulating Estrogen Receptor ß. Front Pharmacol 8:777 (2017). Read more (PubMed: 29163163) »

See all 6 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Mr. Bryan Niedenberger

Verified customer

Submitted Aug 08 2016

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (13%)
Sample
Human Cell lysate - whole cell (HCT116 and MEF cell lysates)
Specification
HCT116 and MEF cell lysates
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Jan 23 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (RAMOS cells)
Specification
RAMOS cells
Treatment
TGF-beta, 24 hours, 50 pM or untreated
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Dec 04 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
African Green Monkey Cell lysate - whole cell (COS-1 cells)
Specification
COS-1 cells
Treatment
TGF-beta, 24 hours, 50 pM or untreated
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Dec 04 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (NMuMG cells)
Specification
NMuMG cells
Treatment
TGF-beta, 24 hours, 50 pM or untreated
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Dec 04 2013

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

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