Anti-alpha Tubulin 抗体 [EP1332Y] - Microtubule Marker (ab52866)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
- Reacts with: Mouse, Rat, Human, Pig, Drosophila melanogaster
Related conjugates and formulations
製品の概要
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製品名
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker
alpha Tubulin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker -
由来種
Rabbit -
特異性
This antibody is expected to recognise most alpha tubulin proteins and not only TUBA4A.
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アプリケーション
適用あり: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human, Pig, Drosophila melanogaster -
免疫原
Synthetic peptide within Human alpha Tubulin aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P68366 -
ポジティブ・コントロール
- WB- HeLa, HEK-293, HepG2, Caco2, NIH/3T3, PC-12, RAW 264.7, PC-12, C6 Jurkat and HEK-293T whole cell lysates; human fetal kidney lysate; Mouse and rat brain lysate; Pig skeletal muscle lysates; IHC-P: Pig kidney tissue; rat kidney tissue; mouse kidney tissue; human breast cancer and stomach tissue; IHC-Fr: Rat kidney tubule tissue; Flow Cyt (intra): HepG2 cells; ICC/IF: HUVEC, HeLa and 293 cells.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1332Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Alexa Fluor® 488 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab185031)
- Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab190573)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272)
- PE Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab208752)
- Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
- Alexa Fluor® 555 Anti-alpha Tubulin antibody [EP1332Y] (ab275113)
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Compatible Secondaries
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Isotype control
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab52866の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | (7) |
1/250 - 1/500.
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Flow Cyt (Intra) |
1/20 - 1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (19) |
1/1000 - 1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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特記事項 |
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ICC/IF
1/250 - 1/500. |
Flow Cyt (Intra)
1/20 - 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the tubulin family. -
翻訳後修飾
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.
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. -
細胞内局在
Cytoplasm > cytoskeleton. - Information by UniProt
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参照データベース
- 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 -
別名
- Alpha-tubulin 1 antibody
- ALS22 antibody
- B ALPHA 1 antibody
see all
画像
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
The negative controls are as follows:
1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution. -
All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/10000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates
Lane 2 : PC-12 (Rat adrenal gland heochromocytoma) whole cell lysates
Lane 3 : NIH/3T3( Mouse embryonic fibroblast)whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling alpha Tubulin with ab52866 at followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human stomach.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
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Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell)cells labelling alpha Tubulin with ab52866 at 1/2000 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730)isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/5000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : C6 (Rat glial tumor cell line) whole cell lysates
Lane 3 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysates
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysates
Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed HepG2 (human liver hepatocellular carcinoma cell line) cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)Hotte, K. et al Biol Open. 2017 Jun 15;6(6):723-731. doi: 10.1242/bio.022335 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/3.0
AJAP1 co-localizes with microtubules in HUVECs
The association of AJAP1 with microtubules in HUVECs is lost upon microtubule destruction. Treatment with 12.5 µM nocodazole for 24 h shows destruction of the microtubule network and loss of AJAP1 tubular localization. For a negative control, HUVECs are treated with DMSO for 24 h. Cell nuclei were counterstained with DAPI (cyan). Microscope: Zeiss LSM 780; objective lens: 63×/1.40 oil; scale bar: 25 µm.
Incubated overnight at 4°C with ab52866.
(From Figure 3E of Hotte et al)
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All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/20000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 50 kDa -
Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/50000 dilution + Rat brain lysates at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)This image is courtesy of an anonymous Abreviewab52866 staining alpha Tubulin in 293 Human embryonic kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 2 hours at 23°C. Samples were incubated with primary antibody (1/200 in 0.5% saponin) for 2 hours at 23°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/50000 dilution + Human fetal kidney lysates at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa -
All lanes : Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (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 ab52866 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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Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) at 1/5000 dilution + Pig skeletal muscle lysates at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 50 kDa
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (240)
ab52866 は 240 報の論文で使用されています。
- Paul S et al. NAK-associated protein 1/NAP1 activates TBK1 to ensure accurate mitosis and cytokinesis. J Cell Biol 223:N/A (2024). PubMed: 38059900
- Gorbacheva EY et al. The Oxidative Phosphorylation and Cytoskeleton Proteins of Mouse Ovaries after 96 Hours of Hindlimb Suspension. Life (Basel) 13:N/A (2023). PubMed: 38137934
- Nowicka H et al. ISGF3 and STAT2/IRF9 Control Basal and IFN-Induced Transcription through Genome-Wide Binding of Phosphorylated and Unphosphorylated Complexes to Common ISRE-Containing ISGs. Int J Mol Sci 24:N/A (2023). PubMed: 38139463
- Huang SS et al. Gut Microbiota-Derived Tryptophan Metabolite Indole-3-aldehyde Ameliorates Aortic Dissection. Nutrients 15:N/A (2023). PubMed: 37836434
- Chang HA et al. YAP nuclear translocation induced by HIF-1α prevents DNA damage under hypoxic conditions. Cell Death Discov 9:385 (2023). PubMed: 37863897