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- Table of Contents
NEIL1 / Endonuclease 8-like 1 · Western blot design guide
Design a Western Blot for NEIL1
Source-linked NEIL1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NEIL1 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
NEIL1 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 | ~43.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked NEIL1 Western Blot Protocol Options
The A02953 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02953; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected NEIL1 Western Blot Band Size?
NEIL1 is predicted at 43.7 kDa; no empirical band is supplied, and the listed features do not demonstrate altered migration.
| Band near 43.7 kDa in whole-cell lysate | Consistent with the predicted size of NEIL1; confirm its identity. |
| Band near 43.7 kDa in a nuclear fraction | Consistent with NEIL1's nuclear location; confirm its identity. |
| Band near 43.7 kDa in a cytoplasmic fraction | Consistent with NEIL1's cytoplasmic location; confirm its identity. |
| Band near 43.7 kDa in a chromosome-enriched fraction | Consistent with NEIL1's chromosome association; confirm its identity. |
💡Expected NEIL1 appearanceUniProt predicts NEIL1 at 43.7 kDa, but no empirical band size is supplied; verify a band near that size with an ordinary identity control.
| UniProt predicted molecular mass | Places the expected band near 43.7 kDa. |
| 390-residue sequence | Underlies the predicted mass; no separate migration effect is established. |
| Nuclear location | May affect fraction recovery; no size change is established. |
| Cytoplasmic location | May affect fraction recovery; no size change is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | NEIL1 recovery from the sampled cellular compartment may be low. | Check loading and examine nuclear and cytoplasmic fractions. |
| Band higher than expected | The higher band's identity is unconfirmed. | Compare with NEIL1 knockdown or knockout material. |
| Band lower than expected | The lower band's identity is unconfirmed. | Compare with NEIL1 knockdown or knockout material. |
| Multiple bands | The supplied features do not establish distinct NEIL1 band sizes. | Use NEIL1 depletion to identify specific bands. |
| Weak or no signal | Recovery or detection of NEIL1 may be insufficient. | Check sample loading, antibody performance, and fraction recovery. |
Experiment evidence
Western blot images showing NEIL1 band
A02953 · NEIL1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NEIL1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NEIL1 in Western blot, you can use appendix tissue lysate, which HPA rates High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside.
⚠️Feasibility: NEIL1 is intracellular, and HPA identifies adipose tissue as a feasible negative tissue control.
HPA tissue expression evidence for NEIL1
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Liver | hepatocytes | High | 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 → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Placenta | decidual cells | Low | Protein (IHC) | HPA → |
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced NEIL1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NEIL1, answered from its protein features.
Where should the NEIL1 band appear?
Band shift · The supplied sequence is 390 residues with a predicted mass of 43.7 kDa. Use 43.7 kDa as a reference when examining the blot; no observed band position is supplied.
Could NEIL1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Do listed modifications predict a NEIL1 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features do not predict a modification-related shift, and they cannot establish why an observed band differs from 43.7 kDa.
Does this guide establish induction of NEIL1?
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.
How should transfer be checked for NEIL1?
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 A02953 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 mitotic samples be compared for NEIL1 quantitation?
Quantitation · NEIL1 associates with centrosomes and condensed chromatin during mitosis. Keep fractionation and sample preparation consistent across conditions, and compare corresponding fractions when quantifying bands.
Which fractions may contain NEIL1?
Interpretation · NEIL1 is listed in the nucleus, on chromosomes, and at the cytoplasmic centrosome. Consider these locations when choosing fractions; during mitosis, it associates with centrosomes and condensed chromatin.
Compare them with the 43.7 kDa predicted mass, but do not assign an identity from size alone. The supplied record gives no observed band position, alternative sequence, or listed modification site that establishes a cause for additional bands.
Compare them with the 43.7 kDa predicted mass, but do not assign an identity from size alone. The supplied record gives no observed band position, alternative sequence, or listed modification site that establishes a cause for additional bands.
Boster reagents
NEIL1 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-Endonuclease 8-like 1 NEIL1 Antibody
Cat # A02953
The catalog reports one anti-NEIL1 polyclonal antibody, A02953, with stated Human and Mouse reactivity. Its WB image caption describes analysis of cells after SDS-PAGE, but the supplied excerpt does not identify the cells or provide detailed validation conditions.
Which to pick: A02953 is the only listed option and has a WB image. Its stated reactivity covers Human and Mouse; check the full validation details against your planned sample before use.
Source: BosterBio NEIL1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96FI4.
- Human Protein Atlas. NEIL1 tissue expression.
- PMC2829949 — target-verified WB comparison
- PMC11387677 — target-verified WB comparison
- PMC4558407 — target-verified WB comparison