This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ZNF23 / Zinc finger protein 23 · Western blot design guide
Design a Western Blot for ZNF23
Source-linked ZNF23 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ZNF23 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
ZNF23 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 | ~73.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked ZNF23 Western Blot Protocol Options
The A13364-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A2780 (40ug), The Testis of Mouse(40ug), Hela (40ug), The Testis of Rat(40ug) (catalog A13364-1) |
| 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 | A13364-1; 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 ZNF23 Western Blot Band Size?
ZNF23 is predicted at 73.1 kDa; two isoforms could affect migration, but no empirical band size or distinct isoform pattern is established.
| Single band near 73.1 kDa | Consistent with the predicted ZNF23 mass; confirm band identity with controls |
| Two bands at different positions | Could reflect isoforms 1 and 2; their migration has not been established |
| Only one detectable band | Does not rule out either annotated isoform |
| Band away from 73.1 kDa | Could reflect an isoform, but its identity requires validation |
💡Expected ZNF23 appearanceUniProt predicts ZNF23 at 73.1 kDa, but no empirical band size or isoform migration is supplied; validate any candidate band with appropriate identity controls.
| UniProt predicted mass | Places the reference band near 73.1 kDa |
| Isoform 1 | Its individual apparent size is not supplied |
| Isoform 2 | Its individual apparent size is not supplied |
| Alternative splicing | May change band position, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ZNF23 may be poorly recovered | Check extraction and compare a nuclear-enriched sample |
| Band higher than expected | Band identity or isoform migration is uncertain | Check antibody specificity with ZNF23 depletion |
| Band lower than expected | An isoform or unrelated band is possible | Test whether the band decreases after ZNF23 depletion |
| Multiple bands | Two isoforms are annotated, but their migration is unknown | Compare bands after ZNF23 depletion |
| Weak or no signal | Nuclear target recovery or abundance may be low | Verify nuclear extraction and sample loading |
Experiment evidence
Western blot images showing ZNF23 band
A13364-1 · ZNF23
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ZNF23 Western blot
🧪For positive controls for ZNF23 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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA expression data are absent, so tissue controls require experimental validation.
HPA tissue expression evidence for ZNF23
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 ZNF23 Western Blot Tips
Deeper troubleshooting and optimisation questions for ZNF23, answered from its protein features.
How should ZNF23 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.
How might ZNF23 isoforms appear on a Western blot?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks canonical residues 1–58, so it may migrate below isoform 1. The features do not establish whether the two bands can be resolved or detected by a particular antibody.
Choose an epitope outside canonical residues 1–58, which are missing from isoform 2. An antibody targeting that segment would not be expected to detect isoform 2. These coordinates use the supplied UniProt canonical sequence.
Choose an epitope outside canonical residues 1–58, which are missing from isoform 2. An antibody targeting that segment would not be expected to detect isoform 2. These coordinates use the supplied UniProt canonical sequence.
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 ZNF23?
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 ZNF23?
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 A13364-1 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 ZNF23 bands be quantified across samples?
Quantitation · Define whether the measurement covers isoform 1, isoform 2, or both. Isoform 2 lacks canonical residues 1–58, so an antibody against that region cannot measure both isoforms. Compare the same assigned band or set of bands across samples.
Why might a ZNF23 band differ from 73.1 kDa?
Interpretation · 73.1 kDa is the predicted mass for the 643-residue canonical protein. The supplied features do not establish an apparent Western-blot mass or demonstrate a band shift. Check whether the band could represent isoform 2, which lacks residues 1–58.
UniProt lists Ubl conjugation and isopeptide bond keywords, but supplies no modified-residue coordinates. These annotations alone do not establish a visible shift or explain a higher band. Treat such a band as unassigned without further evidence.
ZNF23 is annotated as nuclear. Include a nuclear fraction when evaluating detection, and account for the fraction used when comparing band intensity across samples.
First consider the two annotated isoforms and whether the antibody recognizes residues 1–58. The Ubl conjugation keyword permits a modification hypothesis but does not identify a modified site or establish a shifted band. No empirical apparent band is supplied for comparison.
UniProt lists Ubl conjugation and isopeptide bond keywords, but supplies no modified-residue coordinates. These annotations alone do not establish a visible shift or explain a higher band. Treat such a band as unassigned without further evidence.
ZNF23 is annotated as nuclear. Include a nuclear fraction when evaluating detection, and account for the fraction used when comparing band intensity across samples.
First consider the two annotated isoforms and whether the antibody recognizes residues 1–58. The Ubl conjugation keyword permits a modification hypothesis but does not identify a modified site or establish a shifted band. No empirical apparent band is supplied for comparison.
Boster reagents
ZNF23 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-ZNF23 (Q185) Antibody
Cat # A13364-1
The catalog reports A13364-1, an anti-ZNF23 antibody with stated human, mouse, and rat reactivity. Its WB image shows A2780 and HeLa whole-cell lysates and mouse and rat testis lysates, each at 40 µg, using a 1:500 antibody dilution. These are tested examples.
Which to pick: A13364-1 is the only listed ZNF23 antibody and has a WB image. Choose it when its stated reactivity and the shown lysate contexts fit your experiment; the image does not establish performance in every sample type.
Source: BosterBio ZNF23 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P17027.
- Human Protein Atlas. ZNF23 tissue expression.
- PMC8221328 — target-verified WB comparison
- PMC3348897 — target-verified WB comparison