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: Mouse, Rat, Chicken, Human, Zebrafish, Chinese hamster
- 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
Suitable for: WB, ICC/IF, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Chicken, Human, Zebrafish, Chinese hamster
Predicted to work with: Pig, Xenopus laevis
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Immunogen
Synthetic peptide corresponding to Human beta Tubulin aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab20775) -
General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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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Applications
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.
| Application | Abreviews | 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
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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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Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)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.
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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. -
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)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.
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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
References (702)
ab6046 has been referenced in 702 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 ?-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
