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  1. Link

    products/primary-antibodies/alpha-tubulin-antibody-dm1a-bsa-and-azide-free-ab264493.pdf

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Tags & Cell Markers Subcellular Markers Cytoskeleton Microtubules
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Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

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Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Flow Cytometry - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Western blot - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

Key features and details

  • Mouse monoclonal [DM1A] to alpha Tubulin - BSA and Azide free
  • Suitable for: WB, IHC-Fr, Flow Cyt, Electron Microscopy, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 680 Alexa Fluor® 790 FITC HRP

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Overview

  • Product name

    Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free
    See all alpha Tubulin primary antibodies
  • Description

    Mouse monoclonal [DM1A] to alpha Tubulin - BSA and Azide free
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Guinea pig, Hamster, Cow, Dog, Pig, Xenopus laevis, Gerbil, African green monkey
  • Immunogen

    Full length native protein (purified) corresponding to Chicken alpha Tubulin.

  • Epitope

    aa 426-450
  • Positive control

    • WB: HeLa, HEK293, HepG2, Caco2, NIH3T3, PC12, SV40LT-SMC Flow Cytometry: methanol fixed/Tween permeabilised HeLa cells. ICC/IF: Caco-2, NIH3T3, SV40LT-SMC. IHC-P: Human colon, Rat colon.
  • General notes

    ab264493 is the carrier-free version of ab7291.

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

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.40
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    DM1A
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Microtubules
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • Tubulin
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 680 Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab184093)
    • Alexa Fluor® 790 Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab184577)
    • Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195887)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • HRP Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab40742)
    • FITC Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab64503)
    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Related Products

    • alpha Tubulin peptide (ab195584)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab264493 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
1/5000 - 1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

We recommend diluting ab7291 to 1:10000 and incubating overnight at 4°C. Works under both reducing and non-reducing conditions. We recommend using 3% BSA as the blocking agent, blocking with milk may cause a reduction in signal intensity.

IHC-Fr
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Electron Microscopy
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 0.5 - 1 µg/ml.
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/5000 - 1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

We recommend diluting ab7291 to 1:10000 and incubating overnight at 4°C. Works under both reducing and non-reducing conditions. We recommend using 3% BSA as the blocking agent, blocking with milk may cause a reduction in signal intensity.

IHC-Fr
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Electron Microscopy
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 0.5 - 1 µg/ml.
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 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.
  • Target information above from: UniProt accession P68366 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7277 Human
    • Entrez Gene: 22145 Mouse
    • Entrez Gene: 316531 Rat
    • Omim: 191110 Human
    • SwissProt: P68366 Human
    • SwissProt: P68368 Mouse
    • SwissProt: Q5XIF6 Rat
    • Unigene: 75318 Human
    • Unigene: 1155 Mouse
    • Unigene: 92961 Rat
    see all
  • 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

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    ab7291 staining alpha-Tubulin in Caco-2 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab7291 at 1μg/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    IHC image of ab7291 staining alpha Tubulin in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7291, 5ug/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291)

     

  • Flow Cytometry - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Flow Cytometry - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    Overlay histogram showing HeLa cells stained with ab7291 (red line). The cells were fixed with 80% methanol (5 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 (ab7291, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was an anti-mouse DyLight® 488 (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Western blot - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Western blot - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

    Lane 1 : HeLa cell lysate
    Lane 2 : PC12 cell lysate
    Lane 3 : SV40LT-SMC cell lysate
    Lane 4 : NIH/3T3 cell lysate
    Lane 5 : Rat liver tissue lysate
    Lane 6 : Rat heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 50 kDa



    Merged signal (red and green). Green - ab7291 observed at 52 kDa. Red - loading control, ab181602, observed at 38 kDa.

    All samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab7291 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/1,000 and 1/20,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291)

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    ab7291 staining alpha Tubulin in NIH3T3 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab7291 at 1μl/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    FABP4 (green) was detected using FABP4 primary antibody (ab92501; diluted 1/1000). Alpha tubulin (red) was detected using the mouse monoclonal (ab7291) antibody. Cells were imaged by confocal microscopy, using z-stack for adipocyte-like cells.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    IHC image of ab7291 staining alpha Tubulin in rat colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7291, 0.5ug/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    ICC/IF image of ab7291 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab7291, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 WGA was used to label plasma membranes (red).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    ab7291 staining alpha Tubulin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab7291 at a working concentration of 0.5μg/ml and ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    This product also gave a positive signal in 100% methanol (5 min) fixed SV40 cells under the same testing conditions.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291)

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free (ab264493)

    Immunofluorescent imaging of human cells (U2OS) with ab7291 reveals a delicate network of alpha-tubulin (green) located exclusively in the cytoplasm. The nucleus is stained blue.

    IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees.

    Unfortunately, due to size constraints for images on our website, we are unable to show the full uncompressed picture.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab7291).

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

Datasheets and documents

  • Datasheet download

    Download

References (0)

Publishing research using ab264493? Please let us know so that we can cite the reference in this datasheet.

ab264493 has not yet been referenced specifically in any publications.

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