Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)
- Datasheet
- References (20)
- Protocols
Overview
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Product nameAnti-beta IV Tubulin antibody [ONS.1A6]
See all beta IV Tubulin primary antibodies -
DescriptionMouse monoclonal [ONS.1A6] to beta IV Tubulin
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Host speciesMouse
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Tested applicationsSuitable for: IHC-P, ICC/IF, IHC-Fr, ELISA, WBmore details
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Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Hamster, Cow
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Immunogen
Synthetic peptide corresponding to the C-terminal sequence, coupled to BSA.
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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.
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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
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
PurityProtein G purified
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ClonalityMonoclonal
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Clone numberONS.1A6
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IsotypeIgG1
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Light chain typekappa
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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 |
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| 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
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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.
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Sequence similaritiesBelongs to the tubulin family.
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DomainThe highly acidic C-terminal region may bind cations such as calcium.
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Post-translational
modificationsSome 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 localizationCytoplasm > cytoskeleton.
- Information by UniProt
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Database links
- Entrez Gene: 540236 Cow
- Entrez Gene: 10382 Human
- Entrez Gene: 22153 Mouse
- Entrez Gene: 29213 Rat
- Omim: 602662 Human
- SwissProt: Q3ZBU7 Cow
- SwissProt: P04350 Human
- SwissProt: Q9D6F9 Mouse
see all -
Alternative names
- Beta 4 antibody
- Beta 4 tubulin antibody
- beta 5 antibody
see all
Images
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![Western blot - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)](https://www.abcam.com/ps/products/11/ab11315/Images/ab11315-171831-ab11315AP1444909.jpg)
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 minutesThis 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)](https://www.abcam.com/ps/products/11/ab11315/Images/beta-IV-Tubulin-Primary-antibodies-ab11315-13.jpg)
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)](https://www.abcam.com/ps/products/11/ab11315/Images/beta-IV-Tubulin-Primary-antibodies-ab11315-12.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)](https://www.abcam.com/ps/products/11/ab11315/Images/beta-IV-Tubulin-Primary-antibodies-ab11315-8.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)This image is courtesy of Xinmei Chen, University of AlbertaImmunofluorescence 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)](https://www.abcam.com/ps/products/11/ab11315/Images/beta-IV-Tubulin-Primary-antibodies-ab11315-10.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-beta IV Tubulin antibody [ONS.1A6] (ab11315)This image is courtesy of Xinmei Chen, University of AlbertaImmunofluorescence 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.
Protocols
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) »
- Duran GE et al. Decreased levels of baseline and drug-induced tubulin polymerisation are hallmarks of resistance to taxanes in ovarian cancer cells and are associated with epithelial-to-mesenchymal transition. Br J Cancer 116:1318-1328 (2017). Read more (PubMed: 28399108) »
