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
Overview
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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
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanIP HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human beta Tubulin aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab20775) -
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.
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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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
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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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Human
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| IHC-P |
Human
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| IP |
Human
|
| WB |
Human
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| All applications |
Mouse
Rat
Chicken
Pig
Xenopus laevis
Zebrafish
Chinese hamster
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| 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)
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| IHC-P | (1) |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| IP | (1) |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
WB
1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). |
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ICC/IF
1/200. (see PMID: 16030258) |
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IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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IP
Use at an assay dependent concentration. |
Target
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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
modificationsSome 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. - Information by UniProt
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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
see all -
Alternative names
- Beta 4 tubulin antibody
- Beta 5 tubulin antibody
- beta Ib tubulin antibody
see all
Images
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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 withHuman beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 6 : A431 Cell lysate withHuman beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 7 : MCF7 Cell lysate withHuman beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 8 : 293 Cell lysate withHuman 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)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).
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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.
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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)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
Datasheets and documents
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SDS download
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Datasheet download
References (769)
ab6046 has been referenced in 769 publications.
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- 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