Recombinant
RabMAb

Anti-NeuN antibody [EPR12763] - Neuronal Marker (Alexa Fluor® 647) (ab190565)

Overview

  • Product nameAnti-NeuN antibody [EPR12763] - Neuronal Marker (Alexa Fluor® 647)
    See all NeuN primary antibodies
  • Description
    Rabbit monoclonal [EPR12763] to NeuN - Neuronal Marker (Alexa Fluor® 647)
  • ConjugationAlexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested applicationsSuitable for: IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human NeuN aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: A6NFN3

  • Positive control
    • This antibody gave a positive signal in the folowing tissue lysates: Mouse Brain; Mouse Cerebellum; Rat cerebellum; Human Fetal brain as well as the following whole cell lysates: 293T, HeLa and SH-SY5Y lysates; SH-SY-5Y cells. ICC/IF - NGF-differentiated PC12 cells and U87-MG cells
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

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    Alternative versions available:

    Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487)

    Anti-NeuN antibody (Alexa Fluor® 488) [EPR12763] - Neuronal Marker (ab190195)

    Anti-NeuN antibody (Alexa Fluor® 568) [EPR12763] - Neuronal Marker (ab207282)

    Anti-NeuN antibody (Biotin) [EPR12763] (ab204681)

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab190565 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
IHC-P 1/50.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

ICC/IF 1/50.

Target

Anti-NeuN antibody [EPR12763] - Neuronal Marker (Alexa Fluor® 647) images

  • ab190565 staining NeuN in NGF-differentiated PC12 cells (7 days). 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 ab190565 at 1/50 dilution (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Alexa Fluor® 488 Goat anti-Mouse secondary (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

     

  • ab190565 staining NeuN in U87-MG 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 ab190565 at 1/50 dilution(shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Alexa Fluor® 488 Goat anti-Mouse secondary (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • IHC image of ab190565 staining in formalin fixed paraffin embedded tissue section of normal human cerebellum.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with ab190565 (1/50) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then counterstained and mounted with SlowFade® Gold Antifade Mountant with DAPI.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is dervied directly from bound ab190565. The separate images of ab190565 and DAPI alone, combined with the merged version of both signals, shows predominant co-localisation of the Alexa Fluor® 647 signal in the nuclei of the cerebellar granule layer.

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

  • IHC image of ab190565 staining in formalin fixed paraffin embedded tissue section of normal rat cerebellum.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with ab190565 (1/50) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then counterstained and mounted with SlowFade® Gold Antifade Mountant with DAPI.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is dervied directly from bound ab190565. The separate images of ab190565 and DAPI alone, combined with the merged version of both signals, shows predominant co-localisation of the Alexa Fluor® 647 signal in the nuclei of the cerebellar granule layer.

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

  • IHC image of ab190565 staining in formalin fixed paraffin embedded tissue section of normal mouse cerebellum.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with ab190565 (1/50) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then counterstained and mounted with SlowFade® Gold Antifade Mountant with DAPI.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is dervied directly from bound ab190565. The separate images of ab190565 and DAPI alone, combined with the merged version of both signals, shows predominant co-localisation of the Alexa Fluor® 647 signal in the nuclei of the cerebellar granule layer.

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

References for Anti-NeuN antibody [EPR12763] - Neuronal Marker (Alexa Fluor® 647) (ab190565)

ab190565 has not yet been referenced specifically in any publications.

Product Wall

Application Flow Cytometry
Sample Rat Cell (Brain: Whole Hippocampal Tissue)
Permeabilization Yes - 0.1% PBS-Tween
Gating Strategy 1. 7-AAD labelled cells (debris on the left, labelled cells right, gated). 2. Shows physical parameters of the cells, 7-AAD positive cells are in green, they predominantly form a population low in forward and side scatter, and these cells were gated and displayed in grey. 3. Isolation of singlets forming a very low forward scatter from doublets gated and displayed in red. 4. Shows total event population with Alexa647 fluorescence on X-axis and 7-AAD fluorescence on Y-axis, cellular debris and autoflourescence can be seen but a distinct population of 647 positive singlets can also be observed and are gated in purple as NeuN positive cells. Ten thousand Neurons were collected for each sample.
Specification Brain: Whole Hippocampal Tissue
Fixation Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 28 2015

Yes, since the same clone with AF 488 conjugated worked in flow cytometry, we would also expect the AF 647 conjugate to work in flow as well. Since we have not specifically validated ab190565 in flow, you would be eligible for financially risk free tes...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 13 2015

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