Anti-Tubulin 抗体 [YL1/2] - Loading Control (ab6160)
Key features and details
- Rat monoclonal [YL1/2] to Tubulin - Loading Control
- Suitable for: WB, ICC/IF, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2a
Related conjugates and formulations
製品の概要
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製品名
Anti-Tubulin antibody [YL1/2] - Loading Control
Tubulin 一次抗体 製品一覧 -
製品の詳細
Rat monoclonal [YL1/2] to Tubulin - Loading Control -
由来種
Rat -
アプリケーション
適用あり: WB, ICC/IF, IHC-P, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Pig, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, a wide range of other species, Mammals, African green monkey -
免疫原
Full length native protein (purified) corresponding to Saccharomyces cerevisiae Tubulin.
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エピトープ
The YL1/2 monoclonal epitope has been mapped to the last 8 residues (GEEEGEEY) at the carboxy terminus of alpha tubulin when tyrosinated (PubMed IDs: 6415068, 6204858). -
ポジティブ・コントロール
- ICC/IF: HeLa cells. IHC-P: Human colon tissue. WB: HeLa, NIH/3T3, BALB/3T3 and PC-12 whole cell lysate. Flow Cyt (Intra): HeLa cells.
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特記事項
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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精製度
Protein G purified -
一次抗体 備考
This antibody can be used as a loading control on Western blots (Allen et al.) and is not detected by anti-mouse Ig secondaries. It has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe(OH) epitope. Under some circumstances this antibody may cross-react with other protein including E. coli rec A and oxidized actin. -
ポリ/モノ
モノクローナル -
クローン名
YL1/2 -
アイソタイプ
IgG2a -
研究分野
関連製品
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Alternative Versions
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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ab6160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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WB | (17) |
1/5000 - 1/10000.
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ICC/IF | (14) |
1/1000. (see PMID: 16230461)
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IHC-P | (2) |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
特記事項 |
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WB
1/5000 - 1/10000. |
ICC/IF
1/1000. (see PMID: 16230461) |
IHC-P
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use 1µg for 106 cells. ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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
Undergoes a tyrosination/detyrosination cycle, the cyclic removal and re-addition of a C-terminal tyrosine residue by the enzymes tubulin tyrosine carboxypeptidase (TTCP) and tubulin tyrosine ligase (TTL), respectively.
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-tubulins 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: 10376 Human
- Entrez Gene: 203068 Human
- Entrez Gene: 7280 Human
- Entrez Gene: 22143 Mouse
- Entrez Gene: 22151 Mouse
- Entrez Gene: 22153 Mouse
- Entrez Gene: 500929 Rat
- Omim: 191130 Human
see all -
別名
- Tubulin beta 2b antibody
- Alpha tubulin antibody
- Alpha-tubulin ubiquitous antibody
see all
画像
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ICC/IF image of ab6160 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol 5 minutes, 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 1 hour to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6160, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab150165 Alexa Fluor® 488 goat anti-rat IgG (H+L) pre-adsorbed, used at a 1/1000 dilution for 1 hour. 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 4% formaldehyde (10 minutes).
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Lanes 1 & 3 : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/5000 dilution
Lanes 2 & 4 : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/10000 dilution
Lanes 1-2 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
Lanes 3-4 : BALB/3T3 whole cell lysate (ab7901)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Rabbit Anti-Rat IgG H&L (HRP) (ab6734) at 1/2000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?
Additional bands at: 17 kDa, 34 kDa, 80 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds -
Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)Loison-Robert et al PLoS One. 2018 Jan 25;13(1):e0190014. doi: 10.1371/journal.pone.0190014. eCollection 2018. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Cytoskeleton and major extracellular matrix proteins in human DPSC (Dental pulp stem cell) were analyzed by immunofluorescence.
Vimentin and tubulin (Panel D, control, E, (BD) and F, (BR)) are shown.
After 7 days in contact with/without the materials (Biodentine (BD) and Bioroot (BR)), coated coverslip cultures were fixed in PBS (pH 7.4) containing 4% paraformaldehyde/5% sucrose for 10 minutes. For detection of intracellular molecules, the cells on the coverslips were permeabilized using 0.5% Triton X-100. To block background staining, cells were treated with PBS containing 1% BSA/1% glycine at 37°C for 20 minutes. Samples were incubated with the primary antibody at 4°C overnight or at 37°C for 2 hours. For double immunostaining, primary antibodies were incubated as above. Samples were then incubated with the appropriate secondary antibodies at 37°C for 1 hour. Cell nuclei were stained using DAPI.
Scale Bar: 100 μm.
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All lanes : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 3 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase Conjugated Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?
Additional bands at: 85 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
IHC image of Tubulin staining in human colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6 for 20 minutes. The section was then incubated with ab6160, 5 µg/ml, for 15 minutes 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab6160 (red line).
The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab6160, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1 µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
プロトコール
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 (544)
ab6160 は 544 報の論文で使用されています。
- Fare CM et al. A minimal construct of nuclear-import receptor Karyopherin-β2 defines the regions critical for chaperone and disaggregation activity. J Biol Chem 299:102806 (2023). PubMed: 36529289
- Zhang Y et al. Secreted PGK1 and IGFBP2 contribute to the bystander effect of miR-10b gene editing in glioma. Mol Ther Nucleic Acids 31:265-275 (2023). PubMed: 36700043
- Audoynaud C et al. RNA:DNA hybrids from Okazaki fragments contribute to establish the Ku-mediated barrier to replication-fork degradation. Mol Cell 83:1061-1074.e6 (2023). PubMed: 36868227
- Carraro M et al. DAXX adds a de novo H3.3K9me3 deposition pathway to the histone chaperone network. Mol Cell 83:1075-1092.e9 (2023). PubMed: 36868228
- Chen P et al. KNO1-mediated autophagic degradation of the Bloom syndrome complex component RMI1 promotes homologous recombination. EMBO J 42:e111980 (2023). PubMed: 36970874