重组Anti-beta III Tubulin抗体[EPR1568Y] (ab68193)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1568Y] to beta III Tubulin
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
-
产品名称
Anti-beta III Tubulin抗体[EPR1568Y]
参阅全部 beta III Tubulin 一抗 -
描述
兔单克隆抗体[EPR1568Y] to beta III Tubulin -
宿主
Rabbit -
特异性
according to BLAST analysis, it is possible that the antibody will cross-react with TBB6 (Q9BUF5), TBB2B (Q9BVA1), TBB2A (Q13885), TBB5 (P07437), TBB4A (P04350), TBB4B (P68371). This cross-reactivity has not been confirmed experimentally. -
经测试应用
适用于: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human beta III Tubulin aa 1-100 (N terminal). The exact sequence is proprietary.
-
阳性对照
- WB: SH-SY5Y, HeLa, Human cerebellum, Mouse brain and Rat brain lysate Flow Cyt(intra): HeLa cells ICC/IF: Neuro-2a cells, Hap1-TUBB3 WT/KO IHC-P: Human tonsil, Mouse and rat testis tissue sections
-
常规说明
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.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR1568Y -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab68193于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
ICC/IF |
1/50.
For the unpurified format use at 1-5 µg/ml |
|
Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For the unpurified format use at 1/1000 dilution |
|
WB |
1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
For the unpurified format use at 1/20000 to 1/100,000 dilution |
|
IHC-P |
1/8000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Fot the unpurified format use at 1/250 to 1/500 dilution |
说明 |
---|
ICC/IF
1/50. For the unpurified format use at 1-5 µg/ml |
Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For the unpurified format use at 1/1000 dilution |
WB
1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). For the unpurified format use at 1/20000 to 1/100,000 dilution |
IHC-P
1/8000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Fot the unpurified format use at 1/250 to 1/500 dilution |
靶标
-
功能
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. TUBB3 plays a critical role in proper axon guidance and mantainance. -
组织特异性
Expression is primarily restricted to central and peripheral nervous system. -
疾病相关
Defects 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. -
序列相似性
Belongs to the tubulin family. -
结构域
The highly acidic C-terminal region may bind cations such as calcium. -
翻译后修饰
Some 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. -
细胞定位
Cytoplasm > cytoskeleton. - Information by UniProt
-
数据库链接
- Entrez Gene: 10381 Human
- Entrez Gene: 22152 Mouse
- Entrez Gene: 246118 Rat
- Omim: 602661 Human
- SwissProt: Q13509 Human
- SwissProt: Q9ERD7 Mouse
- SwissProt: Q4QRB4 Rat
- Unigene: 511743 Human
see all -
别名
- beta 3 tubulin antibody
- beta 4 antibody
- beta-4 antibody
see all
图片
-
Flow cytometry overlay histogram showing wild-type HeLa (green line) and TUBB3 knockout HeLa stained with ab68193 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab68193) (1x 106 in 100μl at 0.04 μg/ml (1/49250)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type HeLa - black line, TUBB3 knockout HeLa - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
-
Anti-beta III Tubulin antibody [EPR1568Y] (ab68193) at 1/50000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa -
All lanes : Anti-beta III Tubulin antibody [EPR1568Y] (ab68193) at 1/50000 dilution (Purified)
Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
Lane 2 : Human cerebellum lysate
Lane 3 : Mouse brain lysate
Lane 4 : Rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa -
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling beta III Tubulin with Purified ab68193 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
Immunocytochemistry analysis of Neuro-2a(Mouse neuroblastoma neuroblast) cells labeling beta III Tubulin with Purified ab68193 at 1:50 dilution (2.2 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling beta III Tubulin with Purified ab68193 at 1:8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling beta III Tubulin with Purified ab68193 at 1:8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling beta III Tubulin with Purified ab68193 at 1:8000 dilution (0.014 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
-
ab68193 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 (5min), 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 1h. The cells were then incubated with ab68193 at 1μg/ml concentration and ab7291, a mouse anti-tubulin antibody, at 1μg/ml overnight at +4°C. This is followed by a further incubation at room temperature for 1h with a goat anti-rabbit IgG Alexa Fluor® 488 (ab150081) at 2μg/ml (shown in green) and a goat anti-mouse IgG Alexa Fluor® 647 (ab150119) at 2μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (17)
ab68193 被引用在 17 文献中.
- Cheng R et al. P2Y2 Receptor Mediated Neuronal Regeneration and Angiogenesis to Affect Functional Recovery in Rats with Spinal Cord Injury. Neural Plast 2022:2191011 (2022). PubMed: 35154311
- Vargova I et al. Long-Term Cultures of Spinal Cord Interneurons. Front Cell Neurosci 16:827628 (2022). PubMed: 35197829
- Mahmoodi N et al. Improving motor neuron-like cell differentiation of hEnSCs by the combination of epothilone B loaded PCL microspheres in optimized 3D collagen hydrogel. Sci Rep 11:21722 (2021). PubMed: 34741076
- Shan W et al. Polycomb group protein Bmi1 is required for the neuronal differentiation of mouse induced pluripotent stem cells. Exp Ther Med 21:619 (2021). PubMed: 33936276
- Mahmoodi N et al. Microtubule stabilizer epothilone B as a motor neuron differentiation agent for human endometrial stem cells. Cell Biol Int 44:1168-1183 (2020). PubMed: 32022385