Product nameAnti-beta III Tubulin antibody [EP1569Y]
See all beta III Tubulin primary antibodies
DescriptionRabbit monoclonal [EP1569Y] to beta III Tubulin
Tested applicationsSuitable for: mIHC, IHC-Fr, ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human, Zebrafish
Synthetic peptide within Human beta III Tubulin aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: Q13509
- ICC/IF: PC-12, HAP1 and HeLa cells. WB: HAP1 cell lysate. HeLa, PC-12 and HEK-293 whole cell lysate. Mouse and rat brain and spinal cord tissue lysate. IHC-P: Human breast carcinoma tissue. mIHC: Human cerebellum tissue Flow Cyt: U-87 MG and HeLa cells. IHC-Fr: Zebrafish retina sections.
A trial size is available to purchase for this antibody.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Anti-beta III Tubulin antibody [EP1569Y] (HRP) (ab190574)
- Anti-beta III Tubulin antibody [EP1569Y] (Alexa Fluor® 647) (ab190575)
- Anti-beta III Tubulin antibody [EP1569Y] (Biotin) (ab195903)
- Anti-beta III Tubulin antibody [EP1569Y] (Alexa Fluor® 594) (ab201740)
- Anti-beta III Tubulin antibody [EP1569Y] (Alexa Fluor® 568) (ab201849)
- Anti-beta III Tubulin antibody [EP1569Y] - Neuronal Marker (Alexa Fluor® 555) (ab202519)
- Anti-beta III Tubulin antibody [EP1569Y] - BSA and Azide free (ab221935)
- Anti-beta III Tubulin antibody [EP1569Y] (Allophycocyanin) (ab224977)
- Anti-beta III Tubulin antibody [EP1569Y] (FITC) (ab224978)
- Anti-beta III Tubulin antibody [EP1569Y] (Phycoerythrin) (ab224979)
Our Abpromise guarantee covers the use of ab52623 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Perform Sodium citrate antigen retrieval (pH 6.0) between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
|WB||1/1000 - 1/10000. Detects a band of approximately 52 kDa.|
|Flow Cyt||1/10 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionTubulin 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. TUBB3 plays a critical role in proper axon guidance and mantainance.
Tissue specificityExpression is primarily restricted to central and peripheral nervous system.
Involvement in diseaseDefects in TUBB3 are the cause of congenital fibrosis of extraocular muscles type 3A (CFEOM3A) [MIM:600638]. A congenital ocular motility disorder marked by restrictive ophthalmoplegia affecting extraocular muscles innervated by the oculomotor and/or trochlear nerves. It is clinically characterized by anchoring of the eyes in downward gaze, ptosis, and backward tilt of the head. Congenital fibrosis of extraocular muscles type 3 presents as a non-progressive, autosomal dominant disorder with variable expression. Patients may be bilaterally or unilaterally affected, and their oculo-motility defects range from complete ophthalmoplegia (with the eyes fixed in a hypo- and exotropic position), to mild asymptomatic restrictions of ocular movement. Ptosis, refractive error, amblyopia, and compensatory head positions are associated with the more severe forms of the disorder. In some cases the ocular phenotype is accompanied by additional features including developmental delay, corpus callosum agenesis, basal ganglia dysmorphism, facial weakness, polyneuropathy.
Sequence similaritiesBelongs to the tubulin family.
DomainThe highly acidic C-terminal region may bind cations such as calcium.
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.
Cellular localizationCytoplasm > cytoskeleton.
- Information by UniProt
- beta 3 tubulin antibody
- beta 4 antibody
- beta-4 antibody
Lane 1: Wild-type Hap1 cell lysate (20 µg)
Lane 2: TUBB3 knockout Hap1 cell lysate (20 µg)
Lane 3: Wild-type HeLa cell lysate (20 µg)
Lane 4: TUBB3 knockout HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52623 observed at 52 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab52623 was shown to react with beta III Tubulin in wild-type HeLa cells. Loss of signal was observed when knockout sample ab263857 was used. Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. ab52623 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ kit.
The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), ab52623 (1/200 dilution) and ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (blue) was used as a nuclear counter stain.
ab52623 staining beta III Tubulin in wild-type HAP1 cells (top panel) and TUBB3 knockout HAP1 cells (bottom panel).
The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% 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 1 hour. The cells were then incubated with ab52623 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green).
Nuclear DNA was labeled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: beta III Tubulin knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52623 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab52623 was shown to specifically react with beta III Tubulin in wild-type cells as signal was lost in beta III Tubulin knockout HAP1 cells. Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. Ab52623 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively.
Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
In vivo, CK1δ T347 is phosphorylated in trans.
D) Phosphorylation of CK1δ at T347 is not dependent on CK1 activity. HEK-293 cells over-expressing wild-type CK1δ were treated with CK1 inhibitors PF670462 (PF670, 1 μM), PF4800567 (PF480, 1 μM), or D4476 (30 μM) for 3 hours, followed by addition for 30 min of either DMSO or 40 nM Calyculin A. Cell lysates were immunoblotted for anti-pT347 or anti-Myc antibodies. CK1 inhibitors block the autophosphorylation-induced mobility shift but do not decrease phosphorylation of T347. Myc-tagged CK1δ indicated by curly brackets. ab52623 lower panel.
ab52623 staining beta III Tubulin in NGF-differentiated PC-12 (Rat adrenal gland pheochromocytoma cell line) cells.
The cells were fixed with 100% methanol (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 ab52623 at 5 μg/ml and ab7291 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
Negative controls: 1– Rabbit primary 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.
ab52623, at a 1/50 dilution, staining class III beta Tubulin in human breast carcinoma tissue using Immunohistochemistry, Paraffin embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Overlay histogram showing U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells stained with ab52623 (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 (ab52623, 1/1000 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1 μg/1x106) used under the same conditions. Unlabeled 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.
This antibody gave a positive signal in U-87 MG cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
IHC-Fr image of Beta III Tubulin staining on zebrafish retina sections using ab52623 (1:100).
The sections were fixed with paraformaldehyde and permeabilized using Triton-X. Antigen retrieval was performed using Tris-HCl. Blocking was perfomed using 5% BSA for 1 hour at 23°C. ab38738 was diluted 1:100 and incubated with the sections for 16 hours at 4°C. The secondary antibody was goat polyclonal to rabbit IgG conjugated to Alexa Fluor®488 (1:1000).
All lanes : Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/1000 dilution
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Brain (Rat) Tissue Lysate
Lane 3 : Spinal Cord (Mouse) Tissue Lysate
Lane 4 : Spinal Cord (Rat) Tissue Lysate
Lane 5 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution
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 ab52623 overnight at 4°C.
Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/10000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
goat anti-rabbit HRP at 1/2000 dilution
Observed band size: 52 kDa why is the actual band size different from the predicted?
Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling beta III Tubulin with ab52623 at 1/20 (red).
Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.
This product has been referenced in:
- Springer NL et al. Obesity-Associated Extracellular Matrix Remodeling Promotes a Macrophage Phenotype Similar to Tumor-Associated Macrophages. Am J Pathol 189:2019-2035 (2019). Read more (PubMed: 31323189) »
- Sanchez-Rodriguez A et al. Recombinant rabbit beta nerve growth factor production and its biological effects on sperm and ovulation in rabbits. PLoS One 14:e0219780 (2019). Read more (PubMed: 31318941) »