Anti-alpha Tubulin antibody - Loading Control (ab4074)

Overview

  • Product nameAnti-alpha Tubulin antibody - Loading ControlSee all alpha Tubulin primary antibodies ...
  • Description
    Rabbit polyclonal to alpha Tubulin - Loading Control
  • SpecificityThis antibody detects a single band in cell lines, corresponding to alpha tubulin.
  • Tested applicationsIHC-P, WB, Flow Cyt, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human
    Predicted to work with: Pig, Xenopus laevis, Chimpanzee, Monkey, Zebrafish, Chinese Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human alpha Tubulin.

    (Peptide available as ab23537.)

  • Positive control
    • This antibody gave a positive signal in HeLa cells in WB and IF.
  • General notes


    This antibody makes an excellent loading control.

Properties

Applications

Our Abpromise guarantee covers the use of ab4074 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-P
WB
Flow Cyt
ICC/IF
  • Application notesICC/IF: Use at a concentration of 1 µg/ml.
    IHC-P: 1/25.
    WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). Can be blocked with alpha Tubulin peptide (ab23537).

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 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
    • Alternative names
      • Alpha-tubulin 1 antibody
      • FLJ30169 antibody
      • H2 ALPHA antibody
      • Testis specific alpha tubulin antibody
      • TUBA1 antibody
      • TUBA1A antibody
      • TUBA2 antibody
      • TUBA3 antibody
      • TUBA3C antibody
      • TUBA4A antibody
      • Tubulin alpha 1 chain antibody
      • Tubulin alpha antibody
      • Tubulin H2 alpha antibody
      • Tubulin, alpha 1a antibody
      • Tubulin, alpha 3c antibody
      • Tubulin, alpha 4a antibody
      see all

    Anti-alpha Tubulin antibody - Loading Control images

    • All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml

      Lane 1 : Mouse 3T3 lysate
      Lane 2 : Rat liver lysate
      Lane 3 : Cow cell lysate
      Lane 4 : Chicken cell lysate
      Lane 5 : CHO lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
      developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 50 kDa
      Observed band size : 50 kDa


      Exposure time : 30 seconds

      ab4074 cross-reacts strongly with Mouse 3T3 lysate and shows weak cross-reactivity with Rat liver, Cow, Chicken and CHO lysates.

    • ICC/IF image of ab4074 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4074, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM 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 phalloidin was used to label F-actin (red). 
    • (Image courtesy of Human Protein Atlas).
      ab4074 staining alpha tubulin in the glandular cells of the endometrium. Paraffin embedded endometrium tissue of a 33 year-old female was incubated with ab4074 (1/25 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab4074 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

      .

    • Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml + whole cell lysate prepared from MCF7 cells at 20 µg

      Secondary
      Goat anti-rabbit IgG-HRP at 1/1000 dilution
      developed using the ECL technique

      Predicted band size : 50 kDa
      Observed band size : 50 kDa


      Exposure time : 5 minutes

      Image courtesy of an anonymous Abreview.

      See Abreview

    • ab4074 staining alpha Tubulin in murine sperm by Immunocytochemistry/ Immunofluorescence.
      Cells were fixed in 2% paraformaldehyde, permeabilized in -20°C methanol for 10 minutes, blocked using 3% BSA for 1 hour at 25°C and then incubated with ab4074 at a 1/300 dilution for 2 hours at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit (H+L) polyclonal used at a1/400 dilution.

      See Abreview

    • ab4074 staining Tubulin in human LGE neural progenitors by Flow Cytometry. Cells were paraformaldehyde and permeabilized. The sample was incubated with the primary antibody (1/500) for 1 hour at 20°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/800) was used as the secondary antibody.
      Gating Strategy: Isotype population (shown in white).

      See Abreview

    References for Anti-alpha Tubulin antibody - Loading Control (ab4074)

    This product has been referenced in:
    • Chiang HS  et al. GEF-H1 controls microtubule-dependent sensing of nucleic acids for antiviral host defenses. Nat Immunol 15:63-71 (2014). Read more (PubMed: 24270516) »
    • Alagoz M  et al. TDP1 deficiency sensitizes human cells to base damage via distinct topoisomerase I and PARP mechanisms with potential applications for cancer therapy. Nucleic Acids Res 42:3089-103 (2014). WB ; Mouse . Read more (PubMed: 24335147) »

    See all 41 Publications for this product

    Product Wall

    Application Flow Cytometry
    Fixation Paraformaldehyde
    Permeabilization Yes - Perm wash buffer
    Sample Human Cell (LGE Neural Progenitors)
    Specification LGE Neural Progenitors
    Gating Strategy isotype population (shown in white)
    Preparation Cell harvesting/tissue preparation method: 5min collagenase followed by 5mins of accutase
    Sample buffer: 5% FBS wash
    Username

    Dr. Adam Johnson

    Verified customer

    Submitted Apr 18 2014

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (10%)
    Sample Mouse Cell lysate - whole cell (Mouse embryonic stem cells)
    Specification Mouse embryonic stem cells
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 09 2013

    Application Western blot
    Sample Mouse Cell lysate - whole cell (mouse fibroblast)
    Loading amount 50 µg
    Specification mouse fibroblast
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 12 2013

    Thank you for your answer and clarification. This is an extremely important piece of information.

    The most widely used loading controls are tubulin, actin and GAPDH. However, some chemicals can alter the expression levels of these proteins so...

    Read More

    Thank your for your enquiry and interest in our products.

    Could you please explain what experimental system you use with the mouse liver samples? Do you apply any drug which may alter protein expression?

    I look forward to hearing fr...

    Read More
    Application Western blot
    Sample Human Cell lysate - whole cell (human fibroblast cell)
    Loading amount 50 µg
    Specification human fibroblast cell
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 28 2012

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this h...

    Read More

    Thank you for contacting Abcam regarding ab4074.


    I am sorry that you have been experiencing some difficulties using different lots of this antibody in your WB. I have reviewed the history of this antibody and we have not received any othe...

    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (MDA-MB-231)
    Specification MDA-MB-231
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.1% Triton X-100
    Blocking step 0.1% BSA + 1% goat serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted May 24 2012

    Thank you for calling us yesterday to report your concerns over the specificity of theanti-alpha tubulin antibody (ab4074) as well as the changes in our vial sizes you have noticed.

    As discussed over the phone, from time totime we do need to ...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"