Overview

  • Product name
    Anti-HuD + HuC antibody [EPR19098]
  • Description
    Rabbit monoclonal [EPR19098] to HuD + HuC
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse HuC aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q60900

  • Positive control
    • WB: His-tagged mouse HuC recombinant protein; His-GST-tagged mouse HuD recombinant protein; Human fetal brain lysate; Mouse and rat brain lysates; Neuro-2a and SH-SY5Y cell lysates. IHC-P: Human cerebral cortex, colon and glioma tissues; Mouse cerebral cortex and stomach tissues; Rat cerebral cortex and colon tissues. IHC-Fr: Mouse cerebral cortex and stomach tissues; Rat cerebral cortex and colon tissues. ICC/IF: SH-SY5Y and Neuro-2a cells. Flow Cyt: SH-SY5Y and Neuro-2a cells. IP: Mouse brain lysate.
  • General notes

    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 ab184267 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 1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 40 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/500.

Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).

ICC/IF 1/500.
Flow Cyt 1/500.
IP 1/30.

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

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

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weakly cytoplasmic staining on neurons of human cerebral cortex is observed [PMID: 22007133].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic and nuclear staining on neurons of mouse cerebral cortex is observed.

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/2000 dilution

    Lane 1 : His-tagged mouse HuC recombinant protein (aa1-367)
    Lane 2 : His-GST-tagged mouse HuD recombinant protein (aa1-385)
    Lane 3 : His-tagged mouse HuR recombinant protein (aa1-360)
    Lane 4 : His-tagged mouse HuB recombinant protein (aa1-360)

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 40 kDa
    Observed band size: 39,68 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 0.5 second.

    Please note that the band around 55kDa in lane 2 is most likely the HuD protein with tags.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling HuC + HuD with ab184267 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • HuC + HuD was immunoprecipitated from 0.35 mg of mouse brain lysate with ab184267 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab184267 at 1/1000 dilution.

    VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: Mouse brain lysate, 10 µg (Input).

    Lane 2: ab184267 IP in mouse brain lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184267 in mouse brain lysate.

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

    Exposure time: 30 seconds.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

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

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Predicted band size: 40 kDa
    Observed band size: 39 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile is consistent with what has been described in the literature (PMID: 9016658; PMID:8535975).

  • All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 40 kDa
    Observed band size: 39 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3: 1 minute; Lane 4-6: 5 seconds.

    The expression profile is consistent with what has been described in the literature (PMID: 9016658; PMID: 8535975).

  • All lanes : Anti-HuD + HuC antibody [EPR19098] (ab184267) at 1/5000 dilution

    Lane 1 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
    Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 40 kDa
    Observed band size: 39 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution Buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weakly cytoplasmic staining on myenteric ganglia of human colon is observed [PMID: 16918730].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weakly cytoplasmic staining on human glioma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weakly cytoplasmic staining on neurons of mouse cerebral cortex is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic and nuclear staining on myenteric ganglia of mouse stomach is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weakly cytoplasmic staining on neurons of rat cerebral cortex is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling HuC + HuD with ab184267 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic and nuclear staining on myenteric ganglia of rat colon is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse stomach tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic and nuclear staining on myenteric ganglia of mouse stomach is observed (white arrow).

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebral cortex tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic and nuclear staining on neurons of rat cerebral cortex is observed.

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat colon tissue labeling HuC + HuD with ab184267 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic and nuclear staining on myenteric ganglia of rat colon is observed.

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed Neuro-2a (Mouse neuroblastoma cell line) cells labeling HuC + HuD with ab184267 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

References

ab184267 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab184267.
Please use the links above to contact us or submit feedback about this product.

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

Sign up