Overview

  • Product name
    Anti-beta Tubulin antibody
    See all beta Tubulin primary antibodies
  • Description
    Rabbit polyclonal to beta Tubulin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Rice
    Predicted to work with: Rat, Chicken, Guinea pig, Dog, Pig, Saccharomyces cerevisiae, Xenopus laevis, Chimpanzee, Sea urchin, Fungi, Xenopus tropicalis
  • Immunogen

    Synthetic peptide within Human beta Tubulin aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q13509

  • Positive control
    • Recombinant Human beta Tubulin protein (ab70187) can be used as a positive control in WB. MAD109 cells, Skin and lung, HeLa cell line

Properties

Applications

Our Abpromise guarantee covers the use of ab15568 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration. PubMed: 19509307

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/200. Antigen retrieval is not essential but may optimise staining.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.

Target

  • Function
    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.
  • Tissue specificity
    Ubiquitously expressed with highest levels in spleen, thymus and immature brain.
  • Involvement in disease
    Cortical dysplasia, complex, with other brain malformations 6
    Skin creases, congenital symmetric circumferential, 1
  • Sequence similarities
    Belongs to the tubulin family.
  • Domain
    The highly acidic C-terminal region may bind cations such as calcium.
  • Post-translational
    modifications
    Some glutamate residues at the C-terminus are polyglutamylated, resulting in polyglutamate chains on the gamma-carboxyl group (PubMed:26875866). Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold (PubMed:26875866).
    Some glutamate residues at the C-terminus are 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). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear.
    Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
  • Cellular localization
    Cytoplasm, cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Beta 4 tubulin antibody
    • Beta 5 tubulin antibody
    • beta Ib tubulin antibody
    • Beta1 tubulin antibody
    • Class I beta tubulin antibody
    • M40 antibody
    • MGC117247 antibody
    • MGC16435 antibody
    • OK/SW cl.56 antibody
    • OK/SWcl.56 antibody
    • TBB5_HUMAN antibody
    • TUBB 1 antibody
    • TUBB 2 antibody
    • TUBB 5 antibody
    • TUBB antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin beta 1 chain antibody
    • Tubulin beta 2 chain antibody
    • tubulin beta 5 chain antibody
    • Tubulin beta chain antibody
    • Tubulin beta class I antibody
    • tubulin beta polypeptide antibody
    • Tubulin beta-5 chain antibody
    see all

Images

  • All lanes : Anti-beta Tubulin antibody (ab15568) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 55 kDa
    Observed band size: 55 kDa

  • ICC/IF image of ab15568 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab15568, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • ab15568 staining beta Tubulin in IMR32 Human neuroblastoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 1% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA containing TBST) for 16 hours at 4°C. A Cy3®-conjugated Goat anti-rabbit IgG polyclonal (1:1000) was used as the secondary antibody. Blue - nuclei (DAPI).

    See Abreview

  • ab15568 staining beta Tubulin in Human lung by Immunohistochemistry (FFPE-sections).

References

This product has been referenced in:
See all 36 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate pH6
Sample
Sea Urchin Tissue sections (Testes)
Specification
Testes
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 06 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Human Cell (IMR32, Human Neuroblatoma)
Specification
IMR32, Human Neuroblatoma
Permeabilization
Yes - 0.3% triton X 100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 08 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Tissue lysate - whole (Brain)
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 26 2013

Answer

Thank you for contacting us and your interest in our products.
We would advise trying the antibody at a dilution of 1 ug/mL for both flow cytometry and ICC staining. This is only an initial recommendation, you may need to increase or decrease this depending on the results observed with your particular samples. More information on how this antibody has been used successfully in these applications can be found from the following references:
Flow Cyt, IP; Human:
Sorice M et al. Raft component GD3 associates with tubulin following CD95/Fas ligation. FASEB J 23:3298-308 (2009). PubMed: 19509307
ICC
Hines JH et al. Asymmetric endocytosis and remodeling of beta1-integrin adhesions during growth cone chemorepulsion by MAG. Nat Neurosci 13:829-37 (2010). PubMed: 20512137
We have some very useful protocol guides which can be reached form the following link:
https://www.abcam.com/index.html?pageconfig=popular_protocols
and a guide where several applications are included:
https://www.abcam.com/index.html?pageconfig=resource&rid=13111
Which you can request as a hard copy.
I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

Read More

Answer

Thank you for your reply.

I have returned the Abreview to you for revision.

The notes section should be at the bottom third of the form.

Please let me know if you have any questions.

Read More

Answer

Thank you for contacting us. I am very glad to hear that you have taken advantage of our 100% Abreview program.

I have found the review which you have submitted and wish to thank you for this. In order to take advantage of the review code, you will need to add the 100% Abreview code to the additional notes section of the review and if possible submit an image of the western blot.

After this information is submitted you may order as normal and reference this code for you free antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for your response. We have predicted cross reactivity with this species, but we have not yet tested in Saccharomyces cerevisiae and did not know what results would occur when using this in yeast. While products used in 'predicted to react' species are not covered by our Abpromise, I can offer you a retroactive 100% Abreview Testing Discount.For UNTESTED species and/or applications, we have established this testing discount program. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. To redeem this code please submit and Abreview for this product in yeast. Please leave this code in the 'additional notes' box. After submitting the Abreview the code will become active, entitling you to one free antibody. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

Read More

Answer

Thank you for that information.

I have just a few more questions regarding this. What species are you testing this product against? Do you run these westerns natively or in a reduced and denatured condition?

If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.

I look forward to your response. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Read More
Application
Western blot
Sample
Neurospora crassa Cell lysate - whole cell (non)
Loading amount
35 µg
Specification
non
Gel Running Conditions
Reduced Denaturing (4-12% gradient)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 21 2011

Application
Western blot
Sample
Sclerotinia sclerotiorum Cell lysate - whole cell (non)
Loading amount
35 µg
Specification
non
Gel Running Conditions
Reduced Denaturing (4-12% gradient)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 13 2011

1-10 of 11 Abreviews or Q&A

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