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, Human, Chinese hamster
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
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, Human, Chinese hamster
Predicted to work with: Chicken, Pig, Xenopus laevis, Zebrafish
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab20775) -
Positive control
- WB: HeLa, A431, MCF7, NIH3T3, PC12 CHO/K1, 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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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
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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (38) |
1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
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| ICC/IF | (11) |
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.
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| Notes |
|---|
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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
Observed band size: 55 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds -
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)All lanes : Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : NIH3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 3 : PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : CHO/K1 (Chinese hamster ovary cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds -
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)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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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 (938)
ab6046 has been referenced in 938 publications.
- Velasco K et al. Functional evaluation of 16 SCHAD missense variants: Only amino acid substitutions causing congenital hyperinsulinism of infancy lead to loss-of-function phenotypes in vitro. J Inherit Metab Dis 44:240-252 (2021). PubMed: 32876354
- Zhang W et al. The potential role of PSMA6 in modulating fat deposition in pigs by promoting preadipocyte proliferation and differentiation. Gene 769:145228 (2021). PubMed: 33096182
- Li Z et al. Potent aneugenicity of 1-methylpyrene in human cells dependent on metabolic activation by endogenous enzymes. Arch Toxicol 95:703-713 (2021). PubMed: 33057863
- Clapper E et al. The regulation of bcr-abl in hypoxia is through the mTOR pathway. Leuk Lymphoma 62:967-978 (2021). PubMed: 33251904
- Giridharan VV et al. Receptor for Advanced Glycation End Products (RAGE) Mediates Cognitive Impairment Triggered by Pneumococcal Meningitis. Neurotherapeutics 18:640-653 (2021). PubMed: 32886341