NEUROD2 / Neurogenic differentiation factor 2 · Western blot design guide

Design a Western Blot for NEUROD2

Source-linked NEUROD2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NEUROD2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.

Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use

Western blot protocol sheet for NEUROD2: expected band ~41.4 kDa, hero antibody A07904-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **NEUROD2** Western blot protocol sheet — expected band ~41.4 kDa, antibody A07904-1, controls and PMC citations. Open the full NEUROD2 WB guide →

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NEUROD2 Western Blot Experimental Design Guide

Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running

	Expected band	~41.4 kDa
	Observed band	~50 kDa
	Gel	5–20% (catalog A07904-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	—
	Caveat	Observed exceeds expected mass
	Gene-set association	MSigDB C7 membership
	Isoform	1 isoform(s)

Section 1

Source-Linked NEUROD2 Western Blot Protocol Options

The A07904-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human 293T, rat C6, mouse Neuro-2a (catalog A07904-1)
Gel %	5–20% (catalog A07904-1)
Load	30 ug; reducing conditions (catalog A07904-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07904-1)
Membrane	nitrocellulose membrane (catalog A07904-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A07904-1)
Primary antibody	A07904-1 · 0.5 μg/mL (catalog A07904-1)
Primary incubation	overnight at 4°C (catalog A07904-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A07904-1)
Secondary incubation	1.5 hour at RT (catalog A07904-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A07904-1)
Detection	ECL (catalog A07904-1)

Section 2

What Is the Expected NEUROD2 Western Blot Band Size?

NEUROD2 is predicted at 41.4 kDa, but antibody QC reports about 50 kDa; the cause of the difference is not established.

What am I looking at on my blot?
Band near 50 kDa	Matches the empirical NEUROD2 band reported in reducing whole-cell lysates; confirm identity with controls
Band near 41.4 kDa	Near the predicted molecular weight; confirm identity with controls
Stronger band in a nuclear fraction	Consistent with NEUROD2 nuclear localization
Weak or absent band in a cytosolic fraction	Consistent with NEUROD2 nuclear localization

💡Expected NEUROD2 appearanceUniProt predicts 41.4 kDa, while antibody QC reports a band near 50 kDa in reducing whole-cell lysates; the cause of the difference is unknown, so confirm identity with appropriate controls.

How each factor affects band size
UniProt predicted molecular weight	Provides a 41.4 kDa reference for NEUROD2
UniProt molecular weight in daltons	Reports the same predicted mass as 41,361 Da
Predicted full-length mass	Does not by itself establish the apparent position on a blot
Predicted mass compared with observed band	The 41.4 kDa prediction is below the empirical band near 50 kDa; the cause is unestablished

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Poor recovery of nuclear NEUROD2 or low abundance	Check nuclear recovery and a positive lysate
Band higher than expected	The reported band is near 50 kDa, above the 41.4 kDa prediction; its cause is unknown	Compare with the reported band and verify identity by knockdown or another antibody
Band lower than expected	No processing event is annotated to explain a smaller band	Check sample degradation and validate band identity
Multiple bands	Additional size-altering forms are not established by the supplied features	Use knockdown and an independent antibody to identify the NEUROD2 band
Weak or no signal	Low NEUROD2 abundance or poor nuclear recovery	Check a positive control, loading, and nuclear recovery
Fragments below expected size	Possible sample degradation; no cleavage products are annotated	Prepare fresh lysate with protease inhibitors and verify specificity

Source: UniProt Q15784 Human Protein Atlas product datasheet (A07904-1)

Experiment evidence

Western blot images showing NEUROD2 band

A07904-1 · NEUROD2

M07904 · NEUROD2

A30499 · NEUROD2

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for NEUROD2 Western blot

🧪For positive controls for NEUROD2 in Western blot, you can use no HPA-supported tissue or cell line from the supplied evidence.

Positive control: No high/medium HPA tissue identified

Negative control: Suggested KO / knockdown lysate

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: NEUROD2 is nuclear, but the supplied HPA data cannot establish suitable positive or negative samples.

HPA tissue expression evidence for NEUROD2

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Higher expression tissues · candidate positive controls from IHC

Tissue	Cell type	Level	Evidence	Source
No high/medium HPA tissues identified in the supplied evidence.

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
No lower-expression tissue rows available in the supplied evidence.

Section 3

Advanced NEUROD2 Western Blot Tips

Deeper troubleshooting and optimisation questions for NEUROD2, answered from its protein features.

How should NEUROD2 band migration be interpreted?

Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.

Could NEUROD2 isoforms explain multiple bands?

Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Check band identity before assigning any band to NEUROD2.

Do listed modifications explain a NEUROD2 band shift?

PTM · The supplied record lists no modified residues or glycosylation sites. It does not support assigning the difference between 41.4 kDa predicted mass and the observed approximately 50 kDa band to a particular modification.

Does this guide establish induction of NEUROD2?

Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.

What transfer method to use for NEUROD2 Western blot?

Transfer · Choose a transfer setup suitable for the observed band near 50 kDa, and check transfer at that position with a membrane stain. The listed features do not specify a required membrane or transfer method.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A07904-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.

How should NEUROD2 bands be quantified?

Quantitation · Quantify the band identified as NEUROD2 near the supplied observed position of 50 kDa. Keep sample fraction and loading control consistent across lanes. Do not add other bands to the measurement without evidence that they contain NEUROD2.

Why is the NEUROD2 band near 50 kDa?

Interpretation · NEUROD2 has a predicted mass of 41.4 kDa, while the supplied observed band is about 50 kDa. The listed features do not establish the cause of this difference. Compare samples with a molecular weight marker and confirm band identity with an appropriate control.

NEUROD2 is annotated as nuclear. A nuclear-enriched sample may help assess its band; compare the same fraction across samples and use a fraction-appropriate loading control.

NEUROD2 interacts with TCF3, TCF4, TCF12, and CDC20, and its DNA binding requires dimerization with another bHLH protein. Those interaction annotations alone do not identify an unexpected Western blot band. Confirm its identity before attributing it to a NEUROD2 complex.

Boster reagents

NEUROD2 Western Blot Antibodies

Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.

Real WB data

Western blot analysis of NEUROD2 using anti-NEUROD2 antibody (A07904-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human 293T whole cell lysates, Lane 2: rat C6 whole cell lysates, Lane 3: mouse Neuro-2a whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NEUROD2 antigen affinity purified polyclonal antibody (Catalog # A07904-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NEUROD2 at approximately 50 kDa. The expected band size for NEUROD2 is at 41 kDa.

Anti-NEUROD2 Antibody Picoband®

Cat # A07904-1

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Real WB data Western blot analysis of NeuroD2 expression in mouse cerebellum cell lysate.

Anti-NeuroD2 Rabbit Monoclonal Antibody

Cat # M07904

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Real WB data Western Blot analysis of various cells using Neuro D2 Polyclonal Antibody.

Anti-NDF2 Antibody

Cat # A30499

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Three the supplier anti-NEUROD2 antibodies list human, mouse, and rat reactivity and have WB images. A07904-1 shows bands in human 293T, rat C6, and mouse Neuro-2a lysates, with a reported band near 50 kDa versus an expected 41 kDa; independent validation is not supplied.

Which to pick: Choose A07904-1 for the most detailed reported WB conditions and named lysates. M07904 shows a mouse cerebellum lysate blot; A30499 reports various cells without naming them. All three list the same species reactivity, so match your sample and assess the observed band.

Source: BosterBio NEUROD2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q15784.
Human Protein Atlas. NEUROD2 tissue expression.
PMC12223602 — target-verified WB comparison
PMC12821249 — target-verified WB comparison
PMC8259939 — target-verified WB comparison