Invitrogen

NeuroD1 Polyclonal Antibody

Product Image Gallery

NeuroD1 Antibody in Immunofluorescence (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 microgram/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 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

Tested Applications

Dilution

ChIP assay (ChIP)

2.5 µg/10^6 cells

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Western Blot (WB)

1:500-1:3000

Product Specifications

Species Reactivity

Human, Mouse

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

1 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. For short-term storage (1-2 weeks), product can be stored at 4°C. For long-term storage, aliquot and store product at -20° C or below, avioiding multiple freeze-thaw cycles.

Target Information

NeuroD1 encodes a member of the NeuroD family of basic helix-loop-helix (bHLH) transcription factors. The protein forms heterodimers with other bHLH proteins and activates transcription of genes that contain a specific DNA sequence known as the E-box. It regulates expression of the insulin gene, and mutations in this gene result in type II diabetes mellitus.

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

Bioinformatics

Protein Aliases: basic helix-loop-helix transcription factor; beta-cell E-box transactivator 2; Beta-cell E-box transcriptional activator 2; BETA2; BHF1; bHLHa3; Class A basic helix-loop-helix protein 3; 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, (Mouse) Q60867

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

Molecular Function: basic helix-loop-helix transcription factor nuclease nucleic acid binding transcription factor