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

概述

  • 产品名称Anti-beta IV Tubulin抗体[ONS.1A6]
    参阅全部 beta IV Tubulin 一抗
  • 描述
    小鼠单克隆抗体[ONS.1A6] to beta IV Tubulin
  • 经测试应用适用于: IHC-P, ICC/IF, IHC-Fr, ELISA, WBmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Chicken, Hamster, Cow
  • 免疫原

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

  • 阳性对照
    • 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.
  • 常规说明

    Alternative versions available:

    Anti-beta IV Tubulin antibody (HRP) [ONS.1A6] (ab204454)

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能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.
  • 序列相似性Belongs to the tubulin family.
  • 结构域The highly acidic C-terminal region may bind cations such as calcium.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-beta IV Tubulin antibody [ONS.1A6] 图像

  • 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
    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.

  • 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.

  • 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.

  • 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.
  • 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.

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

This product has been referenced in:
  • Mori M  et al. Notch3-Jagged signaling controls the pool of undifferentiated airway progenitors. Development 142:258-67 (2015). Mouse . Read more (PubMed: 25564622) »
  • Wang H  et al. Multiple mechanisms underlying acquired resistance to taxanes in selected docetaxel-resistant MCF-7 breast cancer cells. BMC Cancer 14:37 (2014). WB, ICC/IF ; Human . Read more (PubMed: 24447372) »

See all 9 Publications for this product

Product Wall

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

Xinmei Chen

Verified customer

提交于 Dec 03 2012


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