Invitrogen

NeuroD1 Polyclonal Antibody

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NeuroD1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of NeuroD1 was performed using 70% confluent log phase IMR-32. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with NeuroD1 Rabbit Polyclonal Antibody (Product # PA5-78075) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

NeuroD1 Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunohistochemistry (Frozen) analysis of NeuroD1 was performed in frozen-sectioned rat E13.5 brain tissue using NeuroD1 Polyclonal Antibody (Product # PA5-78075) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

NeuroD1 Antibody in Western Blot (WB) — Western blot was performed using Anti-NeuroD1 Polyclonal Antibody (Product # PA5-78075) and a 45 kDa band corresponding to NEUROD1 was observed in cell line tested. Due to glycosylation a band around 150-230 kDa has also been observed. Modified whole cell extracts (1% SDS) (30 µg lysate) of IMR-32 (Lane 1) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

NeuroD1 Antibody in ChIP assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous NeuroD1 protein using Anti-NeuroD1 Antibody: ChIP was performed using Anti-NeuroD1 Rabbit Polyclonal Antibody (Product # PA5-78075, 5 µg) on sheared chromatin from IMR32 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to NEUROD1, JUNB, cMYC and SLIT2 transcription start sites and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Product Details

PA5-78075

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	-
Immunohistochemistry (Frozen) (IHC (F))	1:100-1:1,000	-
Immunocytochemistry (ICC/IF)	5 µg/mL	-
ChIP assay (ChIP)	2.5 µg/10^6 cells	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Carrier-protein conjugated synthetic peptide encompassing a sequence within the center region of mouse NeuroD1. The exact sequence is proprietary.
Conjugate	Unconjugated
Form	Liquid
Concentration	0.47 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 1% BSA, 20% glycerol
Contains	0.025% Proclin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2736208

Product Specific Information

Positive Control: IMR32, IMR32 nuclear extract

Predicted Reactivity: Mouse (100%), Bovine (100%)

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

NeuroD1 is a transcriptional activator that acts as a differentiation factor during neurogenesis. It has been demonstrated to bind to the insulin gene E-box. Efficient DNA binding requires dimerization with another basic helix-loop-helix (bHLH) protein. Defects in NEUROD1 are a cause of maturity onset diabetes of the young type VI (MODY6). MODY6 is a form of non-insulin-dependent diabetes mellitus characterized by an autosomal dominant mode of inheritance, onset during young adulthood and a primary defect in insulin secretion.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Basic helix-loop-helix factor 1; basic helix-loop-helix transcription factor; beta-cell E-box transactivator 2; Beta-cell E-box transcriptional activator 2; Beta2; BHF-1; BHF1; bHLHa3; Class A basic helix-loop-helix protein 3; neuro d1; NeuroD; NeuroD1; neurogenic differentiation 1; Neurogenic differentiation factor 1; neurogenic helix-loop-helix protein NEUROD

Gene Aliases: BETA2; BHF-1; BHLHA3; MODY6; NEUROD; NEUROD1

UniProt ID: (Human) Q13562, (Rat) Q64289, (Mouse) Q60867

Entrez Gene ID: (Human) 4760, (Rat) 29458, (Mouse) 18012

Molecular Function: basic helix-loop-helix transcription factor