Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)

Reviews (2)Q&A (3)References (21)

Overview

  • Product name
    Anti-beta IV Tubulin antibody [ONS.1A6]
    See all beta IV Tubulin primary antibodies
  • Description
    Mouse monoclonal [ONS.1A6] to beta IV Tubulin
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IHC-Fr, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Hamster, Cow
  • Immunogen

    Synthetic peptide corresponding to the C-terminal sequence, coupled to BSA.

  • Positive control
    • WB: HeLa nuclear extract and HeLa, HEK293, K562, NIH3T3 and U20S whole cell lysates. ICC/IF: methanol fixed HeLa cells. IHC-P: Human normal skin tissue.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab11315 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 17575139
ELISA Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 50 kDa).

3% milk is recommended for blocking non-specific protein-protein interactions.

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.
  • Domain
    The highly acidic C-terminal region may bind cations such as calcium.
  • 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.
  • Cellular localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • Beta 4 antibody
    • Beta 4 tubulin antibody
    • beta 5 antibody
    • Dystonia 4 torsion (autosomal dominant) antibody
    • MC1R antibody
    • TBB4_HUMAN antibody
    • TUB B4 antibody
    • TUBB 4 antibody
    • tubb4 antibody
    • TUBB4A antibody
    • TUBB5 antibody
    • Tubulin 5 beta antibody
    • Tubulin beta 3 antibody
    • Tubulin beta 4 antibody
    • Tubulin beta 4 chain antibody
    • Tubulin beta 4A class IVa antibody
    • Tubulin beta 5 antibody
    • Tubulin beta IV antibody
    • Tubulin beta-4 chain antibody
    see all

Images

  • Western blot - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    Western blot - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    All lanes : Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Additional bands at: 52 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    This blot was produced using a 10% 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 3% Milk before being incubated with ab11315 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)

    IHC image of beta IV Tubulin staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11315, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)

    ICC/IF image of ab11315 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab11315 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)This image is courtesy of Xinmei Chen, University of Alberta
    Immunofluorescence analysis of MCF-7 Cells (human breast cancer cell line), staining beta IV Tubulin with ab11315.

    Cells on coverslip were fixed with -20°C Methanol for 5 min before permeabilization with 0.2% Triton X-100 in PBS for 10 min. Cells were incubated with the primary antibody with a dilution 1/50 for 1 hour. After brief washing with PBS, cells were incubated with TRITC conjugated secondary antibody with a dilution 1/50 for 45 min and washed with PBS to be examined under fluorescence microscope.
  • Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
    Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)This image is courtesy of Xinmei Chen, University of Alberta
    Immunofluorescence analysis of MCF-7 Cells (human breast cancer cell line), staining beta IV Tubulin with ab11315.

    Cells on coverslip were fixed with -20°C Methanol for 5 min before permeabilization with 0.2% Triton X-100 in PBS for 10 min. Cells were incubated with the primary antibody with a dilution 1/50 for 1 hour. After brief washing with PBS, cells were incubated with TRITC conjugated secondary antibody with a dilution 1/50 for 45 min and washed with PBS to be examined under fluorescence microscope.

References

This product has been referenced in:
  • Kohanski MA  et al. Solitary chemosensory cells are a primary epithelial source of IL-25 in patients with chronic rhinosinusitis with nasal polyps. J Allergy Clin Immunol N/A:N/A (2018). Read more (PubMed: 29778504) »
  • Luukkainen A  et al. A Co-culture Model of PBMC and Stem Cell Derived Human Nasal Epithelium Reveals Rapid Activation of NK and Innate T Cells Upon Influenza A Virus Infection of the Nasal Epithelium. Front Immunol 9:2514 (2018). Read more (PubMed: 30467502) »
See all 21 Publications for this product

Publishing research using ab11315? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Immunocytochemistry abreview for Anti-beta IV Tubulin antibody [ONS.1A6]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Primary oligodendrocytes from postnatal day 4 mous)
Permeabilization
Yes - 0.5% Triton X 10 minutes
Specification
Primary oligodendrocytes from postnatal day 4 mous
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More

Julia Edgar

Verified customer

Submitted Jan 09 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-beta IV Tubulin [ONS.1A6] antibody

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MCF-7, human breast cancer cell line)
Specification
MCF-7, human breast cancer cell line
Fixative
Methanol
Permeabilization
Yes - 0.2% Triton X-100 for 10 min.
Read More

Xinmei Chen

Verified customer

Submitted Dec 03 2012

Question

I would like to test ab11315 in ICC/IF

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Answer


I am very pleased to hear you would like to accept our offer and test ab11315 in ICC/IF. This code will give you a discount off your next order before the expiration date. To redeem this offer, please submit an Abreview for ICC/IF and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

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Question

I have a question regarding antibody ab11315. I would like to know whether this antibody detects both the beta IVa and beta IVb subtypes of the beta IV tubulin. I know they only differ at 10 positions so I realize there won't be more then one band present but I would like to know if the antibody is specific to only one subtype or if it recognizes both.

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Answer

Thank you for your enquiry. Unfortunately, we do not have more information about this antibody. We will update the on-line datasheet of this product as soon as we get more data. We apologize for any inconvenience caused. You may want to refer to the references that are listed on the online datasheet for more information (there are Pubmed hyperlinks for each). Please contact us again if you have any additional questions.

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Question

I was interested in purchasing your beta IV tubulin antibody, catalog #ab11315. I was wondering if this was supplied with a blocking peptide.

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Answer

Thank you for your email. This antibody is not supplied with a blocking peptide. If you have any additional questions, please do contact us again.

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