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Recombinant

Recombinant Anti-beta II Tubulin antibody [7B9] (ab28035)

Submit a review Q&A (2)References (4)
Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Immunoprecipitation - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
  • Anti-beta II Tubulin antibody [7B9] (ab28035)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [7B9] to beta II Tubulin
  • Suitable for: WB, IP, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-beta II Tubulin antibody [7B9]
    See all beta II Tubulin primary antibodies

  • Description

    Mouse monoclonal [7B9] to beta II Tubulin

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human, mouse and rat brain tissue lysate. U-87 MG, C6, PC-12, Y79 and MDA-MB-231 whole cell lysate. ICC/IF: U-87 MG, Neuro-2a and PC-12 cells. IP: Neuro-2a whole cell lysate.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab28035 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
WB
1/1000. Predicted molecular weight: 50 kDa.
IP
1/30.
ICC/IF
1/50.
Notes
WB
1/1000. Predicted molecular weight: 50 kDa.
IP
1/30.
ICC/IF
1/50.

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

    Ubiquitous.

  • 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. 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.

  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • bA506K6.1 antibody
    • beta 2 tubulin antibody
    • Beta2 antibody
    • class II beta tubulin isotype antibody
    • class IIa beta-tubulin antibody
    • class IIb beta-tubulin antibody
    • Class IVb beta tubulin antibody
    • dJ40E16.7 antibody
    • DKFZp566F223 antibody
    • FLJ98847 antibody
    • M(beta)2 antibody
    • MGC8685 antibody
    • OTTHUMP00000015956 antibody
    • OTTHUMP00000015964 antibody
    • TBB4B_HUMAN antibody
    • TUBB 2 antibody
    • TUBB 2A antibody
    • TUBB 2C antibody
    • TUBB antibody
    • TUBB PARALOG antibody
    • TUBB2 antibody
    • TUBB2A antibody
    • TUBB2B antibody
    • TUBB2C antibody
    • Tubb4b antibody
    • Tubulin beta 2 antibody
    • Tubulin beta 2 chain antibody
    • Tubulin beta 2A antibody
    • Tubulin beta 2A chain antibody
    • Tubulin beta 2B antibody
    • Tubulin beta 2B chain antibody
    • Tubulin beta 2C antibody
    • Tubulin beta polypeptide antibody
    • Tubulin beta polypeptide 2 antibody
    • Tubulin beta polypeptide paralog antibody
    • Tubulin beta-2 chain antibody
    • Tubulin beta-2C chain antibody
    • Tubulin beta-4B chain antibody
    • tubulin, beta 2A class IIa antibody
    • tubulin, beta 2B class IIb antibody
    • tubulin, beta 4B class IVb antibody
    • Tubulin, beta, class IVB antibody
    • Tubulin, beta-4B antibody
    see all

Images

  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    All lanes : Anti-beta II Tubulin antibody [7B9] (ab28035) at 1/1000 dilution

    Lane 1 : Human brain tissue lysate
    Lane 2 : Human heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 50 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative control: Human heart (PMID:20191564).

  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG cells labelling beta II Tubulin with ab28035 at 1/50 dilution, followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in U-87 MG cell line. ab179513 anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (Red). The nuclear counterstain was DAPI (Blue).

    Negative control 1: ab28035 at a 1/500 dilution followed by ab150080 at a 1/500 dilution.

    Negative control 2: ab179513 at a 1/500 dilution followed by ab150113 at a 1/1000 dilution.

  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    All lanes : Anti-beta II Tubulin antibody [7B9] (ab28035) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-mouse IgG for IP (HRP) (ab131368) at 1/1000 dilution

    Predicted band size: 50 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Immunoprecipitation - Anti-beta II Tubulin antibody [7B9] (ab28035)

    beta II Tubulin was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab28035 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab28035 at 1/1000 dilution. mouse IgG for IP (HRP) (ab131368) was used at 1/1000 dilution.

    Lane 1: Neuro-2a whole cell lysate 10µg.

    Lane 2: ab28035 IP in Neuro-2a whole cell lysate. 

    Lane 3: Mouse monoclonal IgG (ab18443) instead of ab28035 in Neuro-2a whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 seconds.

  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    All lanes : Anti-beta II Tubulin antibody [7B9] (ab28035) at 1/1000 dilution

    Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-mouse IgG for IP (HRP) (ab131368) at 1/1000 dilution

    Predicted band size: 50 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: lane 1, 2: 1 second
    lane 3: 3 seconds

  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a cells labelling beta II Tubulin with ab28035 at 1/50 dilution, followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line. ab179513 anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (Red). The nuclear counterstain was DAPI (Blue).

    Negative control 1: ab28035 at a 1/500 dilution followed by ab150080 at a 1/500 dilution.

    Negative control 2: ab179513 at a 1/500 dilution followed by ab150113 at a 1/1000 dilution.

  • Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Western blot - Anti-beta II Tubulin antibody [7B9] (ab28035)
    All lanes : Anti-beta II Tubulin antibody [7B9] (ab28035) at 1/1000 dilution

    Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
    Lane 2 : Y79 (human retinoblastoma retinoblastoma), whole cell lysate
    Lane 3 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 50 kDa


    Exposure time: 180 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)
    Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [7B9] (ab28035)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 cells labelling beta II Tubulin with ab28035 at 1/50 dilution, followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line. ab179513 anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (Red). The nuclear counterstain was DAPI (Blue).

    Negative control 1: ab28035 at a 1/500 dilution followed by ab150080 at a 1/500 dilution.

    Negative control 2: ab179513 at a 1/500 dilution followed by ab150113 at a 1/1000 dilution.

  • Anti-beta II Tubulin antibody [7B9] (ab28035)
    Anti-beta II Tubulin antibody [7B9] (ab28035)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (4)

Publishing research using ab28035? Please let us know so that we can cite the reference in this datasheet.

ab28035 has been referenced in 4 publications.

  • Wang G  et al. Human amniotic epithelial cells regulate osteoblast differentiation through the secretion of TGFß1 and microRNA-34a-5p. Int J Mol Med 41:791-799 (2018). PubMed: 29207015
  • He X  et al. Resveratrol Attenuates Formaldehyde Induced Hyperphosphorylation of Tau Protein and Cytotoxicity in N2a Cells. Front Neurosci 10:598 (2016). PubMed: 28197064
  • Narvi E  et al. Altered TUBB3 expression contributes to the epothilone response of mitotic cells. Br J Cancer 108:82-90 (2013). PubMed: 23321512
  • Moody SA  et al. Developmental expression of a neuron-specific beta-tubulin in frog (Xenopus laevis): a marker for growing axons during the embryonic period. J Comp Neurol 364:219-30 (1996). PubMed: 8788246

Customer reviews and Q&As

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Submit a review Submit a question

1-2 of 2 Abreviews or Q&A

Question

I am sending you the answered questionnaire and attach our images as you suggest.

We would also like you to recommend us suitable secondary antibody against this primary antibody, because ours really seem to be not working. The most suitable for us would be the one, excitable at 561 or 633 nm.

Best regards,

Read More
Answer

Thank you very much for your email.

Firstly I would like to confirm that the Alexa-488 goat anti-mouse secondary antibodyshould be compatible with anti tubulin antibody ab28035 or ab28036as well. It should also be compatible with work on rat/mouse/human cardiomyocytes. Using an anti mouse antibody on mouse tissue can stain the endogenous immunoglobulins and therefore lead to high background. I am however not sure whether in the freshly isolated cardiocytes there would be any immunoglobulins present.
It might can be useful to use a secondary antibody which has been crossabsorbed against rat immunoglobulins, so it would not recognize the endogenous rat immunogloblins. As per your request, here are some suggestions either excitable at 561nm or 633nm:

-Cy5 conjugated: ab6563 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed against many species, including rat https://www.abcam.com/Goat-Mouse-IgG-HL-Cy5--preadsorbed-ab6563.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-Cy5--preadsorbed-ab6563.html).
-APC conjugated; ab130782 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Allophycocyanin), pre-adsorbed only against human https://www.abcam.com/Goat-Mouse-IgG-HL-APC-preadsorbed-ab130782.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-APC-preadsorbed-ab130782.html).
-DyLight 550 conjugated: ab98713 Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 550), pre-adsorbed against human and rat https://www.abcam.com/Goat-Mouse-IgG-Fc-DyLight-550-preadsorbed-ab98713.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-Fc-DyLight-550-preadsorbed-ab98713.html).
-Texas Red conjugated: ab7066 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (TR), pre-adsorbed against many species, including rat https://www.abcam.com/Goat-Mouse-IgG-HL-Texas-Red-preadsorbed-ab7066.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-Texas-Red-preadsorbed-ab7066.html).
-Texas Red conjugated F(ab')2 fragment: ab5884 Goat F(ab')2 polyclonal Secondary Antibody to Mouse IgG - (Fab)'2 (TR), pre-adsorbed against many species, also against rat https://www.abcam.com/Goat-Fab2-Mouse-IgG-Fab2-Texas-Red-preadsorbed-ab5884.html (or use the following: https://www.abcam.com/Goat-Fab2-Mouse-IgG-Fab2-Texas-Red-preadsorbed-ab5884.html).

Theuse ofF(ab')2 fragment will further reduce the background, if there is some background problem due to unspecificFcportion binding of the secondary antibody. I can therefore recommendthe ab5884.

I have also through the protocol and would like to raise the following points in order to understand the procedure better and toprovide recommendationsin regards to the primaryantibody usedwhich might can help to improve the results:

1.) Can you please confirm which antibody you are using, the ab28035 or the ab28036?

2.) For both antibodies, the fixation method for the ICC has been methanol. I would therefore strongly recommend to use also methanol for fixation if this is possible for you. Indeed, PFA and methanol do fix by different mechanism. The PFA fixes by crosslinking covalently the proteins whereas the methanol fixes by precipitating the proteins. The two mechanism can therefore expose different epitopes and work differently for different antibodies. As we have only tested methanol so far, I can recommend to test also methanol.

3.) I can also recommend to reduce the fixation time for PFA to 15 minutes for primary cell cultures, as overfixation can also reduce the staining.

4.) I can also recommend to test different blocking substances, such as gelatin and serum (of the species in which the secondary antibody has been raised) as alternatives to the BSA to see whether this reduces the background.

5.) In order to reduce the background, I can also recommend to use PBS with 0.05% Tween in the washing buffer. This will help to wash the slides.

I hope this information are helpful. Please do let me know your thoughts and comments. I am looking forward to hear back from you.

Read More

Question

Dear Sir/Madam,

we have ordered Anti-beta II Tubulin antibody [7B9] (ab28035) from your company for immunocytochemestric uses. We have faced some issues and doubts about the labeling mouse cardiomyocytes with this antibody. Could you tell us, please, tell us if there is positive control for this antibody?

Best regards,

Read More
Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products.

As a positive control I can recommend the HeLa cells. Indeed, this antibody has been tested on humantissue so farand on the datasheet you will find a HeLa cell image. https://www.abcam.com/beta-II-Tubulin-antibody-7B9-ab28035.html (or use the following: https://www.abcam.com/beta-II-Tubulin-antibody-7B9-ab28035.html). The antibody has not yet been tested in immunohistochemistry nor on mouse tissue.We do therefore not know whether this antibody can workon this samples, it might well be though.

I would be pleased tohelp you to troubleshoot and optimise the protocol ifyou wish. I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results.

I look forward to receiving your reply. Please do let us know also if you have any other question.

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