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- Table of Contents
NKD1 / Protein naked cuticle homolog 1 · Western blot design guide
Design a Western Blot for NKD1
Source-linked NKD1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NKD1 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
NKD1 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 | ~52.3 kDa | |
| Observed band | ~52 kDa | |
| Gel | 5–20% (catalog A06747-1) | |
| Positive control | Smooth muscle (IHC candidate; verify WB) | |
| Negative control | Adrenal gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked NKD1 Western Blot Protocol Options
The A06747-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human SW620, human Caco-2, human U20S (catalog A06747-1) |
| Gel % | 5–20% (catalog A06747-1) |
| Load | 30 ug; reducing conditions (catalog A06747-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06747-1) |
| Membrane | nitrocellulose membrane (catalog A06747-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06747-1) |
| Primary antibody | A06747-1 · 0.5 μg/mL (catalog A06747-1) |
| Primary incubation | overnight at 4°C (catalog A06747-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06747-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06747-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06747-1) |
| Detection | ECL (catalog A06747-1) |
Section 2
What Is the Expected NKD1 Western Blot Band Size?
NKD1 is predicted at 52.3 kDa and observed near 52 kDa; the cause of the small difference is not established.
| Single band near 52 kDa in whole-cell lysate | Consistent with the reported NKD1 band |
| Band near 52 kDa in a membrane fraction | Consistent with NKD1's annotated cell-membrane location |
| Band near 52 kDa in a cytoplasmic fraction | Consistent with NKD1's annotated cytoplasmic location |
| Band near 52 kDa in both fractions | Consistent with NKD1's two annotated locations |
💡Expected NKD1 appearanceNKD1 is predicted at 52.3 kDa and has an empirical band near 52 kDa in reducing whole-cell blots; confirm band identity with antibody and knockdown controls.
| UniProt predicted molecular weight | Places the expected band near 52.3 kDa |
| UniProt mass in daltons | Gives the same predicted size as 52,285 Da |
| Predicted mass as a band reference | Provides a comparison for the observed band near 52 kDa |
| Predicted mass and empirical migration | Differ by about 0.3 kDa; the cause is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | NKD1 may be below detection or incompletely recovered | Check a positive-control lysate and assess membrane-fraction recovery |
| Band higher than expected | Band identity is uncertain; no listed feature establishes a larger form | Check size markers and whether NKD1 knockdown removes the band |
| Band lower than expected | Possible degradation or nonspecific antibody binding | Use protease inhibitors and test whether NKD1 knockdown removes the band |
| Multiple bands | The supplied record lists no alternative isoforms to explain them | Identify which band decreases after NKD1 knockdown |
| Weak or no signal | Low NKD1 abundance or incomplete recovery | Compare with a positive-control lysate and check fraction recovery |
Experiment evidence
Western blot images showing NKD1 band
A06747-1 · NKD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NKD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NKD1 in Western blot, you can use smooth muscle tissue, which has medium HPA expression.
Positive control: Smooth muscle (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: A clean tissue negative is available, though the positive tissue has only medium HPA expression.
HPA tissue expression evidence for NKD1
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 |
|---|---|---|---|---|
| Smooth muscle | smooth muscle cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NKD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NKD1, answered from its protein features.
Where should the main NKD1 band appear?
Band shift · NKD1 has a predicted mass of 52.3 kDa, close to the supplied observed band of approximately 52 kDa. Use that region to identify the candidate band; mass agreement alone does not confirm its identity.
Could NKD1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to NKD1 isoforms on this evidence alone.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of NKD1?
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 NKD1?
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 A06747-1 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 NKD1 be quantified across fractions?
Quantitation · Quantify the same candidate band near 52 kDa in each sample. Because NKD1 is annotated at both the cell membrane and in the cytoplasm, normalize within each fraction before comparing its distribution.
Does the Myristate annotation predict a band shift?
Interpretation · NKD1 has a Myristate keyword, but the supplied features specify no modified residue or coordinate. This annotation alone does not establish a visible shift or explain a band at a different mass.
NKD1 is annotated at the cell membrane and in the cytoplasm. Examine both fractions if fractionation is part of the experiment, and identify the candidate band near 52 kDa in each.
Compare them with the predicted 52.3 kDa mass and the observed band near 52 kDa. The supplied record lists one isoform and no modified residues or glycosylation sites, so it does not support assigning a different band to an isoform or a specific modification.
NKD1 is annotated at the cell membrane and in the cytoplasm. Examine both fractions if fractionation is part of the experiment, and identify the candidate band near 52 kDa in each.
Compare them with the predicted 52.3 kDa mass and the observed band near 52 kDa. The supplied record lists one isoform and no modified residues or glycosylation sites, so it does not support assigning a different band to an isoform or a specific modification.
Boster reagents
NKD1 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-NKD1 Antibody Picoband®
Cat # A06747-1
The catalog reports one anti-NKD1 antibody for Western blotting: rabbit polyclonal A06747-1, reported reactive with human samples. Its WB image shows an approximately 52 kDa band in HepG2, SW620, Caco-2, and U20S whole-cell lysates. Broader sample validation is not supplied.
Which to pick: A06747-1 is the only listed option. Choose it for human NKD1 Western blots when its documented whole-cell lysate conditions are relevant; the supplied image demonstrates a 52 kDa band in four named human cell lines.
Source: BosterBio NKD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q969G9.
- Human Protein Atlas. NKD1 tissue expression.
- PMC3118196 — target-verified WB comparison
- PMC4882592 — target-verified WB comparison
- PMC5738441 — target-verified WB comparison