Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1332Y] to alpha Tubulin - BSA and Azide free
  • Suitable for: ICC, WB, IHC-P, Flow Cyt
  • Reacts with: Mouse, Rat, Human, Drosophila melanogaster

Overview

  • Product name

    Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free
    See all alpha Tubulin primary antibodies

  • Description

    Rabbit monoclonal [EP1332Y] to alpha Tubulin - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB- HeLa, HEK-293, HepG2, Caco2, NIH/3T3, PC-12, RAW 264.7, PC-12, C6 Jurkat and HEK-293T whole cell lysates; human fetal kidney lysate; Mouse and rat brain lysate; Pig skeletal muscle lysates; IHC-P: Pig kidney tissue; rat kidney tissue; mouse kidney tissue; human breast cancer and stomach tissue; IHC-Fr: Rat kidney tubule tissue; Flow cyt: HepG2 cells; ICC/IF: HUVEC, HeLa and 293 cells.

  • General notes

    ab216650 is the carrier-free version of ab52866. 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

    ab216650 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

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

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 488). Please refer to ab185031 for protocol details.

    ab185031 staining alpha-Tubulin in HeLa 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 ab185031 at a working dilution of 1 in 100 overnight at +4°C (shown in green). Alexa Fluor® 350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410).

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650) This image is courtesy of an anonymous Abreview.
    ab52866 staining alpha Tubulin in 293 Human embryonic kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 2 hours at 23°C. Samples were incubated with primary antibody (1/200 in 0.5% saponin) for 2 hours at 23°C. An Alexa Fluor®555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650) Image from Hotte K. et al Biol Open. 2017 Jun 15;6(6):723-731. doi: 10.1242/bio.022335.

    AJAP1 co-localizes with microtubules in HUVECs

    The association of AJAP1 with microtubules in HUVECs is lost upon microtubule destruction. Treatment with 12.5 µM nocodazole for 24 h shows destruction of the microtubule network and loss of AJAP1 tubular localization. For a negative control, HUVECs are treated with DMSO for 24 h. Cell nuclei were counterstained with DAPI (cyan). Microscope: Zeiss LSM 780; objective lens: 63×/1.40 oil; scale bar: 25 µm.

    Incubated overnight at 4°C with ab52866.

    (From Figure 3E of Hotte et al)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

  • Flow Cytometry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Flow Cytometry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Flow cytometry analysis of 2% paraformaldehyde fixed HepG2 (human liver hepatocellular carcinoma cell line) cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 647). Please refer to ab190573 for protocol details.

    ab190573 staining alpha Tubulin in HeLa 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 ab190573 at a working dilution of 1 in 100 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

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

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Clone EP1332Y (ab216650) has been successfully conjugated by Abcam. This image was generated using Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (PE). Please refer to ab208752 for protocol details.

    ab208752 staining alpha Tubulin in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208752 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52866).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunocytochemistry - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    This ICC data was generated using the same anti-alpha Tubulin antibody clone, EP1332Y, in a different buffer formulation (ab52866).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

    This IHC data was generated using the same anti-alpha Tubulin antibody clone, EP1332Y, in a different buffer formulation (cat# ab52866).

    Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.

    Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
    Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)

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