Anti-Tubulin antibody [YL1/2] - Loading Control

• 产品名称

  Anti-Tubulin抗体[YL1/2] - Loading Control
  参阅全部 Tubulin 一抗

• 描述

  大鼠单克隆抗体[YL1/2] to Tubulin - Loading Control

• 宿主

  Rat

• 经测试应用

  适用于: WB, ICC/IF, IHC-P, Flow Cyt (Intra)more details

• 种属反应性

  与反应: Mouse, Rat, Human
  预测可用于: 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.

• 表位

  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).

• 阳性对照

• 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.

• 常规说明

  
  

  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

• 形式

  Liquid

• 存放说明

  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.

• 存储溶液

  pH: 7.40
  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  Protein G purified

• Primary antibody说明

  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.

• 克隆

  单克隆

• 克隆编号

  YL1/2

• 同种型

  IgG2a

The Abpromise guarantee

Abpromise™承诺保证使用ab6160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。

• 数据库链接

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• Omim: 602530 Human

• Omim: 615101 Human

• SwissProt: P07437 Human

• SwissProt: P68363 Human

• SwissProt: Q13885 Human

• SwissProt: P05213 Mouse

• SwissProt: Q7TMM9 Mouse

• SwissProt: Q9D6F9 Mouse

• SwissProt: Q6P9V9 Rat

• Unigene: 524390 Human

• Unigene: 392113 Mouse

• Unigene: 99661 Rat

• 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

• 别名

• beta Ib tubulin antibody

• CDCBM5 antibody

• CDCBM6 antibody

• fd02b12 antibody

• K ALPHA 1 antibody

• M40 antibody

• OK/SW-cl.56 antibody

• TBA1B_HUMAN antibody

• TUBA1B antibody

• TUBB1 antibody

• TUBB2 antibody

• TUBB5 antibody

• tubulin alpha 1b antibody

• Tubulin alpha-1B chain antibody

• Tubulin alpha-ubiquitous chain antibody

• Tubulin beta 1b antibody

• Tubulin beta 2A antibody

• tubulin beta 2A class IIa antibody

• Tubulin beta antibody

• tubulin beta chain antibody

• tubulin beta class I antibody

• tubulin beta-1 chain antibody

• tubulin beta-2A chain antibody

• tubulin beta-5 chain antibody

• Tubulin K-alpha-1 antibody

• tubulin, alpha, ubiquitous antibody

• tubulin, beta 2A class IIa antibody

• tubulin, beta polypeptide 2 antibody

• tubulin, beta polypeptide antibody

• tubulin, beta, class IIA antibody

• wu:fd02b12 antibody

• zgc:55461 antibody

• Tubulin beta 2b antibody

• Alpha tubulin antibody

• Alpha-tubulin ubiquitous antibody

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

  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).

   

• 

  Western blot - Anti-Tubulin antibody [YL1/2] (ab6160)

  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 kDawhy 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.

• 

  Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

  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 kDawhy 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

• 

  Flow Cytometry (Intracellular) - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

  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.