Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

Overview

• Product nameAnti-alpha Tubulin antibody [DM1A] - Loading ControlSee all alpha Tubulin primary antibodies ...
• Description
  Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control
• SpecificityDM1A causes the 10 nm filaments to collapse into large lateral aggregates collecting in the cell periphery or tight juxtanuclear caps. It does not block microtubule assembly. It does not inhibit polymerisation or depolymerisation of platelet tubulin in vitro. It blocks (by 70-80%) the ability of tubulin dimers (with GppNHp bound) to promote a stable inhibition of adenylyl cyclase. See references for further information on the above.
• Tested applicationsFlow Cyt, ICC/IF, IP, IHC-P, IHC-Fr, Electron Microscopy, WB more details
• Species reactivity
  Reacts with: Mouse, Rat, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Xenopus laevis, Gerbil, African Green Monkey
  
• Immunogen

  Full length native protein (purified) of Chicken alpha Tubulin (extracted from brain).

• Epitopeaa 426-450
• Positive controlIn Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HeLa cells.
• General notes

  
  

  Excellent as a protein loading control antibody.

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS
• Concentration information loading...
• PurityIgG fraction
• Primary antibody notes Excellent as a protein loading control antibody.
• Clonality Monoclonal
• Clone numberDM1A
• IsotypeIgG1
• Light chain typekappa
• Research Areas
  • Signal Transduction
  • Cytoskeleton / ECM
  • Cytoskeleton
  • Microtubules
  • Tubulin

  • Isotype/Loading Controls
  • Loading Controls
  • Tubulin

  • Tags & Cell Markers
  • Subcellular Markers
  • Cytoskeleton
  • Microtubules

Applications

Target

• RelevanceMicrotubules are involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintenance of cell shape. Tubulin itself is a globular protein which consists of two polypeptides (alpha and beta tubulin). Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter. Tyrosine ligase adds a C-terminal tyrosin to monomeric alpha tubulin. Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated alpha tubulin is referred to as Glu-tubulin. Another post-translational modification of detyrosinated alpha tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle. Alpha tubulin is not suitable as a loading control in adipose tissue as expression of tubulin in adipose tissue is very low ( Spiegelman and Farmer, Cell, 1982, 29(1): 53-60, "in cells undergoing adipose differentiation actin synthesis decreases by 90%").
• Cellular localizationMajor constituent of Microtubules
• Database links
  • Entrez Gene: 429035 Chicken
  • Entrez Gene: 504244 Cow
  • Entrez Gene: 7277 Human
  • Entrez Gene: 7846 Human
  • Entrez Gene: 7278 Human
  • Entrez Gene: 22142 Mouse
  • Entrez Gene: 64158 Rat
  • Entrez Gene: 399313 Xenopus laevis

  • Omim: 191110 Human
  • SwissProt: Q5E986 Cow
  • SwissProt: P68366 Human
  • SwissProt: Q71U36 Human
  • SwissProt: Q13748 Human
  • SwissProt: P68369 Mouse
  • SwissProt: P68370 Rat
  • SwissProt: Q4V850 Xenopus laevis

  see all

• Alternative names
  Alpha-tubulin 1 antibodyFLJ30169 antibodyH2 ALPHA antibody

  Testis specific alpha tubulin antibodyTUBA1 antibodyTUBA1A antibodyTUBA2 antibodyTUBA3 antibodyTUBA3C antibodyTUBA4A antibodyTubulin alpha 1 chain antibodyTubulin H2 alpha antibodyTubulin, alpha 1a antibodyTubulin, alpha 3c antibodyTubulin, alpha 4a antibody

  see all

Anti-alpha Tubulin antibody [DM1A] - Loading Control images

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  Western blot - alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
  
  
  Predicted band size : 50 kDa
  
• 

  Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

  ICC/IF image of ab7291 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab7291, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

• 

  Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

  Immunofluorescent imaging of human cells (U2OS) with ab7291 reveals a delicate network of alpha-tubulin (green) located exclusively in the cytoplasm. The nucleus is stained blue.

  IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees.
  

  Unfortunately, due to size constraints for images on our website, we are unable to show the full uncompressed picture in all its glory!

• 

  Western blot - alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
  Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 0.5 µg/ml + Hela cell lysate
  
  Secondary
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6789) at 1/5000 dilution
  developed using the ECL technique
  
  Performed under non-reducing conditions.
  
  Predicted band size : 50 kDa
  Observed band size : 57 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 30 seconds
• 

  Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody [DM1A] - Loading Control (ab7291)This image is courtesy of an anonymous Abreview

  ab7291 at 1/400 staining MCF-10A cells (human mammary epithelial cell line) by ICC/IF. The cells were methanol fixed, blocked with BSA and incubated with the antibody for 16 hours. An Alexa-Fluor ® 488 conjugated goat anti-mouse antibody was used as the secondary. The image shows alpha tubulin staining in green and DAPI counterstain in blue.

  See Abreview

• 

  Flow Cytometry-Anti-alpha Tubulin antibody [DM1A] - Loading Control(ab7291)
  Overlay histogram showing HeLa cells stained with ab7291 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab7291, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
  IF image of ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7291, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% formaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.