Recombinant
RabMAb

Recombinant Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] - BSA and Azide free (ab209348)

Overview

  • Product name

    Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] - BSA and Azide free
    See all alpha Tubulin primary antibodies
  • Description

    Rabbit monoclonal [EPR16772] to alpha Tubulin (acetyl K40) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive).

  • Positive control

    • WB: HeLa, C6 and NIH/3T3 whole cell lysates (treated with 500 ng/ml Trichostatin A for 4 hours); Mouse brain, kidney and spleen lysates; Rat brain and heart lysates; Human fetal heart and fetal kidney lysates. IHC-P: Human and Mouse cerebral cortex tissue; rat cerebellum tissue. IF: HeLa cells treated with 50 ug/ml Trichostatin A for 4 hours. Flow: HeLa cells treated with 500ng/ml Trichostatin A for 4 hours. IP: HeLa treated with 500 ng/ml Trichostatin A for 4 hours.
  • General notes

    Ab209348 is the carrier-free version of ab179484. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab209348 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209348 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 Use at an assay dependent concentration. Detects a band of approximately 52 kDa (predicted molecular weight: 50 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376-Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Sequence similarities

    Belongs to the tubulin family.
  • 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.
    Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links

  • Alternative names

    • Alpha-tubulin 1 antibody
    • ALS22 antibody
    • B ALPHA 1 antibody
    • bA408E5.3 antibody
    • H2 ALPHA antibody
    • Hum a tub1 antibody
    • Hum a tub2 antibody
    • LIS3 antibody
    • MGC171407 antibody
    • MGC55332 antibody
    • TBA4A_HUMAN antibody
    • Testis-specific alpha-tubulin antibody
    • TUBA1 antibody
    • TUBA1A antibody
    • tuba1l antibody
    • Tuba4a antibody
    • Tubulin alpha 1 chain antibody
    • Tubulin alpha antibody
    • Tubulin alpha-1 chain antibody
    • tubulin alpha-1B chain antibody
    • Tubulin alpha-4A chain antibody
    • Tubulin H2-alpha antibody
    • Tubulin, alpha 1 (testis specific) antibody
    • tubulin, alpha 1, like antibody
    • Tubulin, alpha 4a antibody
    • Tubulin, alpha, testis-specific antibody
    • Tubulin, alpha-1 antibody
    see all

Images

  • Ab179484 staining alpha Tubulin in NIH/3T3 (mouse embryonic fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing midbody (arrows) staining in NIH/3T3 cells treated with starvation for 48 hours.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • Ab179484 staining alpha Tubulin in HFF-1 (Human skin fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing cilia (arrows) staining in HFF-1 cells treated with starvation for 48 hours.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • ab179484 stained in Hela cells. Untreated and Trichostatin A treated (50ug/ml, 4 hours) cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab179484 at 1/500 dilution overnight at +4°C. The secondary antibody was ab150177 used at 1 ug/ml for 1hour at room temperature (colored green). DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells treated with 500 ng/ml Trichostatin A for 4 hours labeling alpha Tubulin (acetyl K40) with ab179484 at 1/240 dilution (red line). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.  ab179484 preincubated with 1mg/ml acetyl Alpha tubulin (acetyl K40) peptide (green) or non-acetyl Alpha tubulin (acetyl K40) peptide (orange). The isotype control was Rabbit monoclonal IgG (black) and the unlabelled contol was cells without incubation with primary antibody and secondary antibody (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • Alpha Tubulin was immunoprecipitated from 1mg of HeLa cells (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours with ab179484 at 1/70 dilution. Western blot was performed from 10 µg of the immunoprecipitate using ab179484 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: Hela whole cell extract. Right lane: PBS instead of Hela whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on neuron cells of Mouse cerebral cortex tissue. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on neuron cells of Human brain tissue. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179484).

  • This IHC data was generated using the same anti-alpha Tubulin (acetyl K40) antibody clone, EPR16772, in a different buffer formulation (cat# ab179484).

    Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on Purkinje cells of cerebellum. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • This ICC/IF data was generated using the same anti-alpha Tubulin (acetyl K40) antibody clone, EPR16772, in a different buffer formulation (cat# ab179484).

    Ab179484 staining alpha Tubulin in NIH/3T3 (mouse embryonic fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence).The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing cilia (arrows) staining in NIH/3T3 cells treated with starvation for 48 hours.

References

ab209348 has not yet been referenced specifically in any publications.

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