Recombinant Anti-NeuroD1 antibody [EPR17084] (ab205300)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17084] to NeuroD1
- Suitable for: WB, ICC/IF, IHC-Fr
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-NeuroD1 antibody [EPR17084]
See all NeuroD1 primary antibodies -
Description
Rabbit monoclonal [EPR17084] to NeuroD1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, IHC-Frmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human cerebellum lysate; Y79 whole cell lysate. ICC/IF: SH-SY5Y, Y79 and mouse primary neural/glia cells. IHC-Fr: Frozen mouse brain tissue sections and mouse hippocampus.
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General notes
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17084 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab205300 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 |
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WB |
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 40 kDa).
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ICC/IF |
1/100 - 1/250.
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IHC-Fr |
Use a concentration of 10 µg/ml.
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Notes |
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WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 40 kDa). |
ICC/IF
1/100 - 1/250. |
IHC-Fr
Use a concentration of 10 µg/ml. |
Target
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Function
Differentiation factor required for dendrite morphogenesis and maintenance in the cerebellar cortex. Transcriptional activator. Binds to the insulin gene E-box. -
Involvement in disease
Defects in NEUROD1 are the cause of maturity-onset diabetes of the young type 6 (MODY6) [MIM:606394]. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease. -
Sequence similarities
Contains 1 basic helix-loop-helix (bHLH) domain. -
Post-translational
modificationsPhosphorylated. In islet cells, phosphorylated on Ser-274 upon glucose stimulation; which may be required for nuclear localization. In activated neurons, phosphorylated on Ser-335; which promotes dendritic growth. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4760 Human
- Entrez Gene: 18012 Mouse
- Omim: 601724 Human
- SwissProt: Q13562 Human
- SwissProt: Q60867 Mouse
- Unigene: 574626 Human
- Unigene: 709709 Human
- Unigene: 4636 Mouse
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Alternative names
- atonal antibody
- basic helix loop helix transcription factor antibody
- BETA 2 antibody
see all
Images
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh Mouse hippocampus tissue labelling NeuroD1 with ab205300 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary at 1/1000 dilution. Positive staining on mouse hippocampus tissue is observed. The nuclear counterstain was DAPI (Blue).
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Anti-NeuroD1 antibody [EPR17084] (ab205300) at 1/5000 dilution + Y79 (Human retinoblastoma cell line) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling NeuroD1 with primary antibody anti-NeuroD1 (ab205300) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining in part of mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Anti-MAP2 mouse monoclonal antibody (ab11267) was used to counterstain at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) as a secondary counterstain antibody at 1/1000 dilution. The nuclear counter stain is DAPI (blue).
The negative controls are as follows:-
Negative control 1: ab205300 at 1/100 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
Negative control 2: ab11267 Anti-MAP2 mouse monoclonal antibody at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. -
IHC image of NeuroD1 staining in a section of frozen normal mouse brain performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab205300, 10 ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Y79 (Human retinoblastoma cell line) cells labeling NeuroD1 with ab205300 at 1/250 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 Y79 cell line.
The nuclear counterstain 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:-
-ve control 1: ab205300 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 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. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling NeuroD1 with ab205300 at 1/250 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 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:-
-ve control 1: ab205300 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 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. -
All lanes : Anti-NeuroD1 antibody [EPR17084] (ab205300) at 1/1000 dilution
Lane 1 : Human cerebellum lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh Mouse liver tissue labelling NeuroD1 with ab205300 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary at 1/1000 dilution. No staining on mouse liver tissue is observed. The nuclear counterstain was DAPI (Blue). Negative control (PMID: 24309898).
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Negative tissue image: IHC image of NeuroD1 staining in a section of frozen normal mouse liver performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab205300, 10 ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab205300 has been referenced in 5 publications.
- Seo JH & Jeon YJ Global Proteomic Analysis of Mesenchymal Stem Cells Derived from Human Embryonic Stem Cells via Connective Tissue Growth Factor Treatment under Chemically Defined Feeder-Free Culture Conditions. J Microbiol Biotechnol 32:126-140 (2022). PubMed: 34750284
- Anastasaki C et al. Generation of human induced pluripotent stem cell-derived cerebral organoids for cellular and molecular characterization. STAR Protoc 3:101173 (2022). PubMed: 35199037
- Cejas P et al. Subtype heterogeneity and epigenetic convergence in neuroendocrine prostate cancer. Nat Commun 12:5775 (2021). PubMed: 34599169
- Zhong L et al. HDAC9 Silencing Exerts Neuroprotection Against Ischemic Brain Injury via miR-20a-Dependent Downregulation of NeuroD1. Front Cell Neurosci 14:544285 (2020). PubMed: 33584204
- Dammert MA et al. MYC paralog-dependent apoptotic priming orchestrates a spectrum of vulnerabilities in small cell lung cancer. Nat Commun 10:3485 (2019). PubMed: 31375684