Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
Key features and details
- Rabbit polyclonal to alpha Tubulin - Microtubule Marker
- Suitable for: ICC/IF, Flow Cyt, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
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Overview
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Product name
Anti-alpha Tubulin antibody - Microtubule Marker
See all alpha Tubulin primary antibodies -
Description
Rabbit polyclonal to alpha Tubulin - Microtubule Marker -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Drosophila melanogaster, Indian muntjac, African green monkey, Chinese hamster -
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.
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Positive control
- WB: HeLa, HEK-293, HepG2, Caco-2, NIH/3T3 and PC-12 whole cell lysates. ICC/IF: HeLa, Caco-2, NIH/3T3 and SV40LT-SMC cells.
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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...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Corresponding Antibody
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab18251 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 |
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ICC/IF | (12) |
Use a concentration of 1 - 5 µg/ml.
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Flow Cyt |
Use 0.01µg for 106 cells.
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WB | (6) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
Abcam recommends using milk as the blocking agent. |
Notes |
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ICC/IF
Use a concentration of 1 - 5 µg/ml. |
Flow Cyt
Use 0.01µg for 106 cells. |
WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). Abcam recommends using milk as the blocking agent. |
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. -
Sequence similarities
Belongs to the tubulin family. -
Post-translational
modificationsSome 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.
Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling. -
Cellular localization
Cytoplasm > cytoskeleton. - Information by UniProt
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Database links
- Entrez Gene: 7277 Human
- Entrez Gene: 22145 Mouse
- Entrez Gene: 316531 Rat
- Omim: 191110 Human
- SwissProt: P68366 Human
- SwissProt: P68368 Mouse
- SwissProt: Q5XIF6 Rat
- Unigene: 75318 Human
see all -
Alternative names
- Alpha-tubulin 1 antibody
- ALS22 antibody
- B ALPHA 1 antibody
see all
Images
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All lanes : Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) at 0.5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
Lane 2 : HEK-293 (Human embryonic kidney cell line) whole cell lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) whole cell lysate
Lane 4 : Caco-2 (Human colonic carcinoma cell line) whole cell lysate
Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 6 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab18251 overnight at 4°C. Antibody binding was detected using Anti-Rabbit Alexa Fluor® 790 (ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
ab18251 staining alpha-Tubulin in SV40LT-SMC cells.
The cells were fixed with 100% methanol for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab18251 at 1 μl/ml and ab7291 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with an anti-rabbit Alexa Fluor® 488 (ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
ab18251 staining alpha-Tubulin in NIH3T3 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18251 at 1 μl/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit Alexa Fluor® 488 (ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
ab18251 staining alpha-Tubulin in Caco-2 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18251 at 5 μl/ml and ab7291 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit Alexa Fluor® 488 (ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
ab18251 staining alpha Tubulin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18251 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Microtubule Marker (ab18251)
ab18251 at a 1/8000 dilution staining human HeLa cells by immunocytochemistry. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes. The secondary antibody was a Cy3® conjugated Goat Anti-Rabbit IgG (H+L). The image shows staining of an interphase IM cell.
This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.
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All lanes : Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab18251 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP (ab97051), and visualised using ECL development solution ab133406
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All lanes : Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) at 0.5 µg/ml
Lane 1 : HeLa lysate
Lane 2 : A431 lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Flour Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa
Additional bands at: 30 kDa (possible cross reactivity)ab18251 detects a strong band at 50 kDa corresponding to alpha tubulin. Cross-reactivity is also seen with other lower molecular weight bands. This may be reduced by using the antibody at a lower working concentration.
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Overlay histogram showing NIH3T3 cells stained with ab18251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Overlay histogram showing SV40LT-SMC cells stained with ab18251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Overlay histogram showing Caco2 cells stained with ab18251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (286)
ab18251 has been referenced in 286 publications.
- Feng W et al. SUMOylation of a-tubulin is a novel modification regulating microtubule dynamics. J Mol Cell Biol 13:91-103 (2021). PubMed: 33394042
- Seo HR et al. CHIP and BAP1 Act in Concert to Regulate INO80 Ubiquitination and Stability for DNA Replication. Mol Cells 44:101-115 (2021). PubMed: 33658435
- Kawanishi N & Machida S Alterations of macrophage and neutrophil content in skeletal muscle of aged versus young mice. Muscle Nerve 63:600-607 (2021). PubMed: 33386611
- Eastwood EL et al. Dimerisation of the PICTS complex via LC8/Cut-up drives co-transcriptional transposon silencing in Drosophila. Elife 10:N/A (2021). PubMed: 33538693
- Kunz TC et al. The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy. Front Cell Infect Microbiol 11:644750 (2021). PubMed: 33796486