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- Table of Contents
NEUROD1 / Neurogenic differentiation factor 1 · Western blot design guide
Design a Western Blot for NEUROD1
Real validated NEUROD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NEUROD1 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
NEUROD1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~39.9 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog A01038-2) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated NEUROD1 Western Blot Protocols
The A01038-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human U251, rat C6 (catalog A01038-2) |
| Gel % | 10% (catalog A01038-2) |
| Load | 30 ug; reducing conditions (catalog A01038-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01038-2) |
| Membrane | nitrocellulose membrane (catalog A01038-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01038-2) |
| Primary antibody | A01038-2 · 0.5 μg/mL (catalog A01038-2) |
| Primary incubation | overnight at 4°C (catalog A01038-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01038-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01038-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01038-2) |
| Detection | ECL (catalog A01038-2) |
Section 2
What Is the Expected NEUROD1 Western Blot Band Size?
NEUROD1 is predicted at 39.9 kDa and observed at ~50 kDa; the cause of that difference is not established by the supplied features.
| Band near 50 kDa | Empirical NEUROD1 band in reducing whole-cell blots; its difference from the predicted mass is unexplained |
| Band near 40 kDa | Near the 39.9 kDa sequence-based prediction; confirm its identity |
| Close doublet near the main band | Could reflect different phosphorylation states, but the listed sites do not establish a visible separation |
| Weak band in a cytoplasmic fraction | NEUROD1 can also localize to the nucleus |
💡Expected NEUROD1 appearanceNEUROD1 has a predicted mass of 39.9 kDa, while antibody QC reports a band at ~50 kDa in reducing whole-cell blots; the cause of the difference is unestablished, so confirm band identity with ordinary controls.
| Predicted sequence mass | 39.9 kDa is predicted; the observed band is ~50 kDa, with no established cause for the difference |
| Phosphoserine 162 | May affect mobility; no visible size effect is established |
| Phosphoserine 259 | May affect mobility; no visible size effect is established |
| Phosphoserine 266 | May affect mobility; no visible size effect is established |
| Phosphoserine 274 | May affect mobility; no visible size effect is established |
| Phosphoserine 335 by CaMK2 | May affect mobility; no visible size effect is established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The empirical ~50 kDa band exceeds the 39.9 kDa prediction; its cause is unestablished | Compare with the reported ~50 kDa band and confirm identity using NEUROD1 depletion or a second antibody |
| Band lower than expected | A lower band has no established feature-based assignment | Check the molecular-weight marker and confirm identity with NEUROD1 depletion or a second antibody |
| Multiple bands | Different phosphorylation states are possible, but distinct bands are unproven | Compare bands after phosphatase treatment and confirm their identity with NEUROD1 depletion |
| Weak or no signal | NEUROD1 can localize to the nucleus, affecting its recovery in a cytoplasmic fraction | Compare nuclear and cytoplasmic fractions and check fractionation controls |
| Fragments below expected size | No listed cleavage feature establishes a specific fragment | Check sample integrity and confirm smaller bands with a second antibody or NEUROD1 depletion |
Experiment evidence
Western blot images showing NEUROD1 band
A01038-2 · NEUROD1
M01038 · NEUROD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NEUROD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NEUROD1 in Western blot, you can use bone marrow, which HPA scores as medium expression.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: NEUROD1 is intracellular, and HPA identifies a not-detected tissue, making tissue lysate controls feasible.
HPA tissue expression evidence for NEUROD1
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NEUROD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NEUROD1, answered from its protein features.
How should NEUROD1 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 NEUROD1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no specific isoform explanation for multiple bands. Check band identity before labeling an additional band as an isoform.
Which NEUROD1 phosphorylation sites matter when comparing bands?
PTM · UniProt lists phosphoserines at positions 162, 259, 266, 274, and 335; position 335 is phosphorylated by CaMK2. Record these as UniProt sequence coordinates when comparing antibody or paper numbering. The annotations do not establish which sites are occupied in your sample or whether they change migration.
Can glucose stimulation change where NEUROD1 is detected?
Induction · NEUROD1 is annotated in the cytoplasm and nucleus. In pancreatic islet cells, nuclear shuttling in response to glucose stimulation is noted by similarity. If studying that setting, compare matched nuclear and cytoplasmic fractions and keep the glucose condition consistent.
How should transfer be checked for NEUROD1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01038-2 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 NEUROD1 be quantified across glucose conditions?
Quantitation · For pancreatic islet cells, the supplied localization note suggests glucose may redistribute NEUROD1 between cytoplasm and nucleus, based on similarity. Quantify matched fractions or whole-cell samples consistently; a change in one fraction alone does not establish a change in total abundance.
Why is the NEUROD1 band near 50 kDa instead of 39.9 kDa?
Interpretation · The supplied apparent band is ~50 kDa, while the predicted mass is 39.9 kDa. NEUROD1 has five annotated phosphoserines, but their presence alone does not explain this difference or establish a visible shift. Compare the band with a validated NEUROD1 control before assigning it.
Efficient DNA binding requires dimerization with another bHLH protein, and NEUROD1 can heterodimerize with TCF3/E47. That functional annotation does not identify a high Western-blot band as a dimer. Validate unexpected bands with an independent NEUROD1-specific control.
Efficient DNA binding requires dimerization with another bHLH protein, and NEUROD1 can heterodimerize with TCF3/E47. That functional annotation does not identify a high Western-blot band as a dimer. Validate unexpected bands with an independent NEUROD1-specific control.
Boster reagents
NEUROD1 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
Anti-NEUROD1 Antibody Picoband®
Cat # A01038-2
Real WB data
Anti-NeuroD1 Rabbit Monoclonal Antibody
Cat # M01038
Two the supplier anti-NEUROD1 antibodies have WB images. A01038-2 was shown with human K562 and U251 and rat C6 lysates; its reported band is approximately 50 kDa versus an expected 40 kDa. M01038 was shown with Y79 lysate; its caption gives no band size.
Which to pick: For human or rat samples matching the documented contexts, consider A01038-2. M01038 lists human, mouse, and rat reactivity and has a Y79 lysate WB image, but the supplied caption does not document mouse or rat test samples. Neither image establishes performance in every tissue.
Source: BosterBio NEUROD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13562.
- Human Protein Atlas. NEUROD1 tissue expression.
- PMC4782570 — target-verified WB comparison
- PMC7873949 — target-verified WB comparison
- PMC7772964 — target-verified WB comparison