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    beta-tubulin-antibody-loading-control-ab6046.pdf

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Isotype/Loading Controls Loading Controls Tubulin
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Anti-beta Tubulin antibody - Loading Control (ab6046)

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  • SDS
Reviews (60)Q&A (18)References (769)

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Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
  • Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody - Loading Control (ab6046)

Key features and details

  • Rabbit polyclonal to beta Tubulin - Loading Control
  • Suitable for: WB, ICC/IF, IHC-P, IP
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-beta Tubulin antibody - Loading Control
    See all beta Tubulin primary antibodies
  • Description

    Rabbit polyclonal to beta Tubulin - Loading Control
  • Host species

    Rabbit
  • Specificity

    This antibody detects a single clean band at 50kD representing beta Tubulin. This band is significantly reduced by using peptide blocking.
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human beta Tubulin aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab20775)

  • Positive control

    • WB: HeLa, A431, MCF7, and 293 cell lysates; IP: HeLa whole cell extract; ICC: HeLa cells; IHC-P: Human liver carcinoma tissue section.
  • General notes

      

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Isotype/Loading Controls
    • Loading Controls
    • Tubulin
    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Microtubules
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • Tubulin
    • Isotype/Loading Controls
    • Loading Controls
    • Beta Tubulin

Associated products

  • Alternative Versions

    • HRP Anti-beta Tubulin antibody - Loading Control (ab21058)
    • Anti-beta Tubulin antibody [AA2] (ab231082)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Immunizing Peptide (Blocking)

    • Human beta Tubulin peptide (ab20775)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human beta Tubulin protein (ab70187)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-SDHB antibody [21A11AE7] (ab14714)
    • Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)
    • Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
    • Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
All applications
Mouse
Rat
Chicken
Pig
Xenopus laevis
Zebrafish
Chinese hamster
Application Abreviews Notes
WB (38)
1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
ICC/IF (10)
1/200. (see PMID: 16030258)
IHC-P (1)
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP (1)
Use at an assay dependent concentration.
Notes
WB
1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
ICC/IF
1/200. (see PMID: 16030258)
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.

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.
  • Tissue specificity

    Ubiquitously expressed with highest levels in spleen, thymus and immature brain.
  • Involvement in disease

    Cortical dysplasia, complex, with other brain malformations 6
    Skin creases, congenital symmetric circumferential, 1
  • 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, resulting in polyglutamate chains on the gamma-carboxyl group (PubMed:26875866). Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold (PubMed:26875866).
    Some glutamate residues at the C-terminus are 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). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear.
    Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
  • Cellular localization

    Cytoplasm, cytoskeleton.
  • Target information above from: UniProt accession P07437 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 396254 Chicken
    • Entrez Gene: 203068 Human
    • Entrez Gene: 22154 Mouse
    • Entrez Gene: 733686 Pig
    • Entrez Gene: 29214 Rat
    • Entrez Gene: 380418 Xenopus laevis
    • Entrez Gene: 386701 Zebrafish
    • Omim: 191130 Human
    • SwissProt: P07437 Human
    • SwissProt: P99024 Mouse
    • SwissProt: Q767L7 Pig
    • SwissProt: P69897 Rat
    • SwissProt: Q91575 Xenopus laevis
    • Unigene: 636480 Human
    • Unigene: 46552 Xenopus laevis
    see all
  • Alternative names

    • Beta 4 tubulin antibody
    • Beta 5 tubulin antibody
    • beta Ib tubulin antibody
    • Beta1 tubulin antibody
    • Class I beta tubulin antibody
    • M40 antibody
    • MGC117247 antibody
    • MGC16435 antibody
    • OK/SW cl.56 antibody
    • OK/SWcl.56 antibody
    • TBB5_HUMAN antibody
    • TUBB 1 antibody
    • TUBB 2 antibody
    • TUBB 5 antibody
    • TUBB antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin beta 1 chain antibody
    • Tubulin beta 2 chain antibody
    • tubulin beta 5 chain antibody
    • Tubulin beta chain antibody
    • Tubulin beta class I antibody
    • tubulin beta polypeptide antibody
    • Tubulin beta-5 chain antibody
    see all

Images

  • Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
    Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
    All lanes : Anti-beta Tubulin antibody - Loading Control (ab6046) at 1/500 dilution

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

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa


    Exposure time: 10 seconds
  • Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
    Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
    Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)

    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.

  • Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
    Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody - Loading Control (ab6046)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody - Loading Control (ab6046)
    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.

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (769)

Publishing research using ab6046? Please let us know so that we can cite the reference in this datasheet.

ab6046 has been referenced in 769 publications.

  • Gao J  et al. Metformin protects against PM2.5-induced lung injury and cardiac dysfunction independent of AMP-activated protein kinase a2. Redox Biol 28:101345 (2020). PubMed: 31669973
  • Yin L  et al. Exogenous hydrogen sulfide alleviates surgery-induced neuroinflammatory cognitive impairment in adult mice by inhibiting NO signaling. BMC Anesthesiol 20:12 (2020). PubMed: 31918664
  • Huang Q  et al. Loss of LAMTOR1 in pancreatic ß-cells increases glucose-stimulated insulin secretion in mice. Int J Mol Med 45:23-32 (2020). PubMed: 31939616
  • Massah S  et al. The effects of the DNA Demethylating reagent, 5-azacytidine on SMCHD1 genomic localization. BMC Genet 21:3 (2020). PubMed: 31941450
  • Rhodes JDP  et al. Cohesin Disrupts Polycomb-Dependent Chromosome Interactions in Embryonic Stem Cells. Cell Rep 30:820-835.e10 (2020). PubMed: 31968256
View all Publications for this product

Customer reviews and Q&As

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1-10 of 60 Abreviews

Western blot abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (293T, HepaRG, Huh7, HepG2)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
40 µg
Specification
293T, HepaRG, Huh7, HepG2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Dr. Romain Parent

Verified customer

Submitted Oct 14 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
African green monkey Cell (COS7 cells)
Permeabilization
Yes - Saponin
Specification
COS7 cells
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Oct 05 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549)
Permeabilization
Yes - Triton X-100
Specification
A549
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jun 16 2020

Western blot abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Fish Cell lysate - whole cell (ATCC-CRL-2872)
Gel Running Conditions
Reduced Non-Denaturing (Native) (10% SDS)
Loading amount
1e+006 cells
Specification
ATCC-CRL-2872
Blocking step
BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Feb 10 2020

Western blot abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U-87MG cell line)
Gel Running Conditions
Reduced Denaturing (4-15% gradient polyacrylamide gel)
Loading amount
30 µg
Specification
U-87MG cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Read More

Abcam user community

Verified customer

Submitted Feb 05 2020

Western blot abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Multiple myeloma cancer cell line lysates)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
Multiple myeloma cancer cell line lysates
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jan 15 2020

Western blot abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (striatum)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
striatum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 23°C
Read More

Dr. Alexandre Magno

Verified customer

Submitted Jul 01 2019

Immunohistochemistry free floating abreview for Anti-beta Tubulin antibody - Loading Control

Average
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain
Read More

Abcam user community

Verified customer

Submitted Feb 12 2019

Immunocytochemistry abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (primary mouse astrocytes)
Permeabilization
Yes - 0.1% triton x100
Specification
primary mouse astrocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde
Read More

Qasim Alhadidi

Verified customer

Submitted Nov 27 2018

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-beta Tubulin antibody - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.2% Triton X
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Mr. J. Y. Seo

Verified customer

Submitted Apr 04 2018

1-10 of 60 Abreviews

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