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- Table of Contents
ZFP42 / Zinc finger protein 42 homolog · Western blot design guide
Design a Western Blot for ZFP42
Source-linked ZFP42 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ZFP42 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
ZFP42 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 | ~34.8 kDa | |
| Observed band | ~45 kDa | |
| Gel | 5–20% (catalog A08177-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ZFP42 Western Blot Protocol Options
The A08177-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human placenta, human Hela, human Caco-2, human 22RV1 (catalog A08177-1) |
| Gel % | 5–20% (catalog A08177-1) |
| Load | 50ug; reducing conditions (catalog A08177-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A08177-1) |
| Membrane | Nitrocellulose membrane (catalog A08177-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A08177-1) |
| Primary antibody | A08177-1 · 0.5 μg/mL (catalog A08177-1) |
| Primary incubation | overnight at 4°C (catalog A08177-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A08177-1) |
| Secondary incubation | 1.5 hour at RT (catalog A08177-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A08177-1) |
| Detection | ECL (catalog A08177-1) |
Section 2
What Is the Expected ZFP42 Western Blot Band Size?
ZFP42 is predicted at 34.8 kDa but observed near 45 kDa; the cause of the difference is not established.
| Band near 45 kDa | Empirical ZFP42 band; identity still requires controls |
| Band near 34.8 kDa | Matches the predicted mass, but migration there is unverified |
| Weak band in whole-cell lysate | Nuclear ZFP42 may be easier to detect in a nuclear fraction |
| Multiple bands | The single annotated isoform does not establish their identities |
💡Expected ZFP42 appearanceZFP42 has a predicted mass of 34.8 kDa and an empirical band near 45 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | The reference protein has a calculated mass of 34.8 kDa |
| 310-residue sequence | Defines the reference sequence used for the calculated mass |
| Single annotated isoform | Provides no evidence for distinct isoform sizes |
| Ubl conjugation keyword | Suggests a possible larger conjugated species, but no modified site or band is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ZFP42 may be poorly represented in the tested lysate | Check a nuclear fraction and a positive control |
| Band higher than expected | The observed 45 kDa band exceeds the 34.8 kDa prediction for an unestablished reason | Compare with the reported band and verify identity by ZFP42 depletion |
| Band lower than expected | The band may be unrelated or a fragment; no cleavage feature is supplied | Check antibody specificity and test an independent epitope |
| Multiple bands | The single annotated isoform does not account for distinct bands | Identify which bands respond to ZFP42 depletion |
| Weak or no signal | Nuclear localization may reduce signal in whole-cell lysate | Test a nuclear fraction alongside a positive control |
Experiment evidence
Western blot images showing ZFP42 band
A08177-1 · ZFP42
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ZFP42 Western blot
🧪For positive controls for ZFP42 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: HPA provides no expression data, so suitable positive and negative samples cannot be confirmed.
HPA tissue expression evidence for ZFP42
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 ZFP42 Western Blot Tips
Deeper troubleshooting and optimisation questions for ZFP42, answered from its protein features.
How should ZFP42 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.
Which ZFP42 isoforms should I expect?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning multiple bands to annotated ZFP42 isoforms.
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 ZFP42?
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 ZFP42 Western blot?
Transfer · The supplied features do not specify a transfer method. Assess transfer efficiency across the predicted 34.8 kDa mass and the reported apparent band near 45 kDa using a molecular weight marker and a post-transfer protein stain.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A08177-1 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 I quantify nuclear ZFP42?
Quantitation · ZFP42 is annotated as nuclear. Compare the same verified band across consistently prepared nuclear samples, and normalize to a suitable nuclear loading measure. Keep the band definition consistent if additional bands appear.
Why might ZFP42 appear near 45 kDa instead of 34.8 kDa?
Interpretation · The supplied apparent band is about 45 kDa, while the predicted mass is 34.8 kDa. The listed features do not establish the cause of that difference. Use a molecular weight marker and confirm the band’s identity before attributing the difference to a modification.
Ubl conjugation and isopeptide bond are listed as keywords, but no modified residue is mapped. A higher band could be investigated as a conjugated form; these annotations alone do not establish a visible shift or identify a band.
Check bands against the predicted 34.8 kDa mass and the reported band near 45 kDa. The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. Ubl conjugation is annotated without a mapped modified residue, so band position alone cannot establish an identity.
Ubl conjugation and isopeptide bond are listed as keywords, but no modified residue is mapped. A higher band could be investigated as a conjugated form; these annotations alone do not establish a visible shift or identify a band.
Check bands against the predicted 34.8 kDa mass and the reported band near 45 kDa. The record lists one isoform, no signal peptide or propeptide, and no glycosylation sites. Ubl conjugation is annotated without a mapped modified residue, so band position alone cannot establish an identity.
Boster reagents
ZFP42 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-Rex1/ZFP42 Antibody Picoband®
Cat # A08177-1
The catalog reports A08177-1, a rabbit polyclonal anti-Rex1/ZFP42 antibody with stated human, mouse, and rat reactivity. Its Western blot image shows an approximately 45 kDa band in human placenta, HeLa, Caco-2, and 22RV1 lysates; the supplied image does not demonstrate mouse or rat WB performance.
Which to pick: A08177-1 is the only listed option and has a WB image using human samples. For mouse or rat samples, reactivity is listed, but the supplied WB evidence is limited to human lysates.
Source: BosterBio ZFP42 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96MM3.
- Human Protein Atlas. ZFP42 tissue expression.
- PMC12809756 — target-verified WB comparison
- PMC11655532 — target-verified WB comparison
- PMC7156282 — target-verified WB comparison