Anti-Tubulin Polymerization Promoting Protein antibody [EPR3316] (ab92305)

Overview

  • Product nameAnti-Tubulin Polymerization Promoting Protein antibody [EPR3316]
    See all Tubulin Polymerization Promoting Protein primary antibodies
  • Description
    Rabbit monoclonal [EPR3316] to Tubulin Polymerization Promoting Protein
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Tubulin Polymerization Promoting Protein.

  • Positive control
    • Human normal brain and fetal brain tissue, Human glioma, Human, mouse and rat cerebral cortex; Mouse brain and Rat brain lysates; SH-SY5Y and Neuro-2a cells
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Alternative versions available:

    Anti-Tubulin Polymerization Promoting Protein antibody (Alexa Fluor® 488) [EPR3316] (ab203991)
    Anti-Tubulin Polymerization Promoting Protein antibody (Alexa Fluor® 647) [EPR3316] (ab204011)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
  • Storage bufferpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberEPR3316
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92305 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 1/10000. Predicted molecular weight: 24 kDa.

For unpurified use at 1/500 - 1/1000.

IHC-P 1/50. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/250 - 1/500.

Flow Cyt 1/20 - 1/50.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.

Target

  • FunctionMay play a role in the polymerization of tubulin into microtubules, microtubule bundling and the stabilization of existing microtubules, thus maintaining the integrity of the microtubule network. May play a role in mitotic spindle assembly and nuclear envelope breakdown.
  • Tissue specificityWidely expressed.
  • Sequence similaritiesBelongs to the TPPP family.
  • Post-translational
    modifications
    Poor substrate for GSK3 (By similarity). Phosphorylated by LIMK1 on serine residues. Phosphorylation may alter the tubulin polymerization activity.
  • Cellular localizationCytoplasm. Cytoplasm, cytoskeleton. Nucleus. Localizes to glial Lewy bodies in the brains of individuals with synucleinopathies.
  • Information by UniProt
  • Database links
  • Alternative names
    • 25 kDa brain specific protein antibody
    • 25 kDa brain-specific protein antibody
    • Brain specific protein p25 alpha antibody
    • Glycogen synthase kinase 3 (GSK3) inhibitor p24 antibody
    • OTTHUMP00000161630 antibody
    • p24 antibody
    • p25 antibody
    • p25-alpha antibody
    • p25alpha antibody
    • TPPP antibody
    • TPPP/p25 antibody
    • TPPP_HUMAN antibody
    • TPPP1 antibody
    • Tubulin polymerization promoting protein antibody
    • Tubulin polymerization-promoting protein antibody
    see all

Anti-Tubulin Polymerization Promoting Protein antibody [EPR3316] images

  • All lanes : Anti-Tubulin Polymerization Promoting Protein antibody [EPR3316] (ab92305) at 1/10000 dilution

    Lane 1 : Human cerebellum tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Mouse cerebral cortex tissue lysate
    Lane 4 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size : 24 kDa
    Observed band size : 25 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM /TBST
  • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue sections labelling Tubulin Polymerization Promoting Protein with purified ab92305 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence staining of Neuro-2a (mouse neuroblastoma) cells labelling Tubulin Polymerization Promoting Protein with purified ab92305 at a working dilution of 1/100. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. DAPI was used as nuclear counterstain. The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit secondary (ab150077).

  • Overlay histogram showing 4% paraformaldehyde fixed Neuro-2a (mouse neuroblastoma) cells labelling Tubulin Polymerization Promoting Protein with purified ab92305 at dilution of 1/20. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody. 

  • All lanes : Anti-Tubulin Polymerization Promoting Protein antibody [EPR3316] (ab92305) at 1/1000 dilution

    Lane 1 : Fetal brain lysate
    Lane 2 : SHSY5Y cell lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit antibody at 1/2000 dilution

    Predicted band size : 24 kDa
  • Immunohistochemical analysis of paraffin-embedded human glioma tissue sections labelling Tubulin Polymerization Promoting Protein with purified ab92305 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • ab92305 at 1/100 dilution staining Tubulin Polymerization Promoting Protein in SH-SY5Y cells, by immunofluorescence.
  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling Tubulin Polymerization Promoting Protein with purified ab92305 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue sections labelling Tubulin Polymerization Promoting Protein with purified ab92305 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Overlay histogram showing SH-SY5Y cells stained with ab92305 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab92305, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • ab92305 at 1/250 dilution staining Tubulin Polymerization Promoting Protein in paraffin-embedded Human brain tissue by immunohistochemistry.

References for Anti-Tubulin Polymerization Promoting Protein antibody [EPR3316] (ab92305)

This product has been referenced in:
  • Ray S  et al. LIM kinase regulation of cytoskeletal dynamics is required for salivary gland branching morphogenesis. Mol Biol Cell 25:2393-407 (2014). Read more (PubMed: 24966172) »

See 1 Publication for this product

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