Anti-beta Tubulin抗体- Loading Control (ab6046)

概述

  • 产品名称Anti-beta Tubulin抗体- Loading Control
    参阅全部 beta Tubulin 一抗
  • 描述
    兔多克隆抗体to beta Tubulin - Loading Control
  • 特异性This antibody detects a single clean band at 50kD representing beta Tubulin. This band is significantly reduced by using peptide blocking.
  • 经测试应用适用于: ICC, IHC-Fr, WB, ICC/IF, ELISA, IHC-P, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Human, Pig, Xenopus laevis, Zebrafish, Chinese Hamster
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human beta Tubulin.

    (Peptide available as ab20775.)

  • 阳性对照
    • HeLa Cell lysate; A431 Cell lysate; MCF7 Cell lysate; 293 Cell lysate; HeLa Cell lysate; A431 Cell lysate; MCF7 Cell lysate; 293 Cell lysate;

性能

应用

Our Abpromise guarantee covers the use of ab6046 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
ICC 1/200. (see Abreview)
IHC-Fr 1/200. (see Abreview)
WB 1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
ICC/IF 1/200. (see PMID: 16030258)
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

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.
  • 组织特异性Ubiquitously expressed with highest levels in spleen, thymus and immature brain.
  • 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 tubulin antibody
    • Beta 5 tubulin antibody
    • BetaTubulin antibody
    • TBB5_HUMAN antibody
    • TUBB antibody
    • TUBB2 antibody
    • TUBB2A antibody
    • TUBB5 antibody
    • tubulin beta 2A antibody
    • Tubulin beta chain antibody
    • Tubulin beta-5 chain antibody
    see all

Anti-beta Tubulin antibody - Loading Control 图像


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa


    Exposure time : 10 seconds

    Lane 1 - 8 : beta Tubulin antibody - Loading Control (ab6046) at 1/500 dilution

    Lane 1 : HeLa Cell lysate at 20 ug
    Lane 2 : A431 Cell lysate at 20 ug
    Lane 3 : MCF7 Cell lysate at 20 ug
    Lane 4 : 293 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 5 : HeLa Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 6 : A431 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 7 : MCF7 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 8 : 293 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml

  • ICC/IF image of ab6046 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 (ab6046, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.

  • ab6046 staining beta Tubulin in human stomach tissue by Immunohistochemistry (frozen sections). Tissue was fixed with acetone and then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody at a 1/200 dilution for 1 hour at 23°C. An undiluted HRP-conjugated goat polyclonal was used as secondary antibody.

    See Abreview

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).

  • Adiponectin (green) was detected using adiponectin primary antibody (ab22554; 2.5 µl/mL). Beta tubulin (red) was detected using the rabbit polyclonal (ab6046) antibody. Cells were imaged by confocal microscopy, using z-stack for adipocyte-like cells.

  • Beta Tubulin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Tubulin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab6046.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 50kDa: beta Tubulin.
  • 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 (ab6046, 1µg/ml and ab37266, 1µg/ml) overnight at +4°C. The secondary antibodies were ab150115 Alexa Fluor® 647 goat anti-mouse IgG (H+L) used at 2µg/ml for 1h and ab175652 Alexa Fluor® 405 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. Nuclear Green LCS1 (ab138904) was used to stain the cell nuclei (green) at a dilution of 1/500.

     

  • IHC image of beta Tubulin staining in human liver carcinoma FFPE 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 ab6046, 5µg/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.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa

    Lane 1 - 8 : beta Tubulin antibody - Loading Control (ab6046) at 1/500 dilution

    Lane 1 : HeLa Cell lysate at 20 ug
    Lane 2 : A431 Cell lysate at 20 ug
    Lane 3 : MCF7 Cell lysate at 20 ug
    Lane 4 : 293 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 5 : HeLa Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 6 : A431 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 7 : MCF7 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml
    Lane 8 : 293 Cell lysate at 20 ug with beta Tubulin peptide (ab20775) at 1 ug/ml

    Secondary

Anti-beta Tubulin antibody - Loading Control (ab6046)参考文献

This product has been referenced in:
  • Hedberg ML  et al. Genetic landscape of metastatic and recurrent head and neck squamous cell carcinoma. J Clin Invest 126:169-80 (2016). WB ; Human . Read more (PubMed: 26619122) »
  • Huang R  et al. The role of HDAC2 in chromatin remodelling and response to chemotherapy in ovarian cancer. Oncotarget 7:4695-711 (2016). WB ; Human . Read more (PubMed: 26683361) »

See all 285 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Mr. Bryan Niedenberger

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提交于 Aug 04 2016

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Mouse Cultured Cells (Mouse Embryonic Fibroblasts)
Specification Mouse Embryonic Fibroblasts
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
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提交于 May 26 2014

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Human Cultured Cells (H460)
Specification H460
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
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提交于 May 26 2014

Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (8)
Sample Human Cell lysate - whole cell (MDAMB231)
Specification MDAMB231
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
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提交于 Jan 09 2014

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (10% GEL)
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 Dec 13 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% GEL)
Sample Mouse Cell lysate - whole cell (MEF)
Specification MEF
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 Dec 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Rat Tissue lysate - whole (Mammary Tumor)
Specification Mammary Tumor
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jun 25 2013

We would recommend the use of one of the following products for a loading control with zebrafish samples: AB15224 anti alpha Tubulin (http://www.abcam.com/alpha-tubulin-antibody-ab15246.html). This product has been featured in the following publicat...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Tissue lysate - whole (Head lysates)
Loading amount 30 µg
Specification Head lysates
Gel Running Conditions Reduced Denaturing (12% TGX (Bio-Rad))
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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提交于 Nov 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (vaginal epithelial cells)
Loading amount 40 µg
Specification vaginal epithelial cells
Gel Running Conditions Reduced Denaturing (12% Tris-glycine)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Nov 06 2012

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