Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5526] to Huntingtin - BSA and Azide free
  • Suitable for: IHC-FoFr, IHC-P, WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Huntingtin antibody [EPR5526] - BSA and Azide free
    See all Huntingtin primary antibodies

  • Description

    Rabbit monoclonal [EPR5526] to Huntingtin - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Mouse
    IHC-FoFr
    Mouse
    IHC-P
    Mouse
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: SH-SY5Y, HeLa, HAP1, PC-12 and Neuro-2a whole cell lysates and mouse and rat brain lysates. IHC-P: Human cerebral cortex tissue, Human astrocytoma tissue, Mouse testis and Rat testis tissue. ICC/IF: Neuro-2a and SH-SY5Y cell lines. Flow cytometry: SH-SY5Y cell line. IHC-Fr: Mouse cerebellum tissue

  • General notes

    Ab209668 is the carrier-free version of ab109115. 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

    ab209668 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.

Properties

Images

  • Western blot - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Western blot - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : HTT knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 348 kDa
    Observed band size: 348 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab109115).

      Lanes 1- 2: Merged signal (red and green). Green - ab109115 observed at 348 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab109115 was shown to react with Huntingtin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265976 (knockout cell lysate ab256946) was used. Wild-type HeLa and HTT knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109115 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunohistochemistry (Frozen) analysis of mouse cerebellum tissue sections labeling Huntingtin with purified ab109115 at 1/100 (13.4 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. Sections were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Negative control: PBS instead of the primary antibody. DAPI (blue) was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Flow Cytometry - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Flow Cytometry - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with purified ab109115 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Western blot - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Western blot - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HTT knockout HAP1 whole cell lysate
    Lane 3 : SH-SY5Y whole cell lysate
    Lane 4 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 348 kDa



    This WB data was generated using the same anti-Huntingtin antibody clone, EPR5526, in a different buffer formulation (cat# ab109115).

    Lanes 1 - 4: Merged signal (red and green). Green - ab109115 observed at 348 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab109115 was shown to specifically recognize HTT in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when HTT knockout samples were exmined. Wild-type and HTT knockout samples were subjected to SDS-PAGE. Unpurified ab109115 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10,000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunohistochemical analysis of paraffin-embedded Rat testis labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Weak cytoplasmic staining on spermatogenic cells of rat testis.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

    SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with purified ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    1. ab191472 at 1/1000 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunohistochemistry (Frozen) analysis of mouse cerebrum tissue sections labeling Huntingtin with purified ab109115 at 1/100 (13.4 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. Sections were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Negative control: PBS instead of the primary antibody. DAPI (blue) was used as nuclear counterstain. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) was performed.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide ab109115).

  • Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunocytochemistry/ Immunofluorescence - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

    Neuro-2a (Mouse neuroblastoma cells) cells labeling Huntingtin with purified ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on Neuro-2a cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    1. ab191472 at 1/1000 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunohistochemical analysis of paraffin-embedded Mouse testis labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Cytoplasmic staining on spermatogenic cells of mouse testis.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on neuron of human cerebral cortex was observed.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109115).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

    This IHC data was generated using the same anti-Huntingtin antibody clone, EPR5526, in a different buffer formulation (cat# ab109115).

    Immunohistochemical analysis of paraffin-embedded Human astrocytoma labeling Huntingtin with purified ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on cancer cells of astrocytoma.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)
    Anti-Huntingtin antibody [EPR5526] - BSA and Azide free (ab209668)

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