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- Table of Contents
PADI4 / Protein-arginine deiminase type-4 · Western blot design guide
Design a Western Blot for PADI4
Source-linked PADI4 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PADI4 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
PADI4 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 | ~74.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Spleen (IHC candidate; verify WB) +1 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 PADI4 Western Blot Protocol Options
The PA2043 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | PANC, 293T (catalog PA2043) |
| 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 | PA2043; 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 PADI4 Western Blot Band Size?
PADI4 is predicted at 74.1 kDa; its listed citrullination sites have no demonstrated migration effect, and no empirical band size is supplied.
| Single band near 74.1 kDa | Consistent with the UniProt predicted mass for PADI4; confirm identity with controls |
| Band near 74.1 kDa in a nuclear fraction | Consistent with PADI4 nuclear localization |
| Band near 74.1 kDa in a cytoplasmic fraction | Consistent with PADI4 cytoplasmic localization |
| Band near 74.1 kDa in a granule-enriched fraction | Consistent with PADI4 localization to cytoplasmic granules |
💡Expected PADI4 appearanceUniProt predicts 74.1 kDa for PADI4, but no empirical band size is supplied; a band near that size remains a candidate requiring ordinary identity controls.
| UniProt predicted mass | Places the expected full-length band near 74.1 kDa |
| Citrullination at residue 205 | Documented modification with no demonstrated band-size effect |
| Citrullination at residue 212 | Documented modification with no demonstrated band-size effect |
| Citrullination at residue 218 | Documented modification with no demonstrated band-size effect |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PADI4 abundance in the tested lysate is unknown | Check sample expression and include a known positive lysate |
| Band higher than expected | Identity of the higher band is unestablished | Compare with a PADI4 depletion control |
| Band lower than expected | Identity of the lower band is unestablished | Check sample integrity and compare with a PADI4 depletion control |
| Multiple bands | No alternative isoforms are listed to explain distinct bands | Use a PADI4 depletion control to identify specific bands |
| Weak or no signal | PADI4 may be distributed across cytoplasmic, nuclear, and granule fractions | Check fraction recovery and test a known positive lysate |
Experiment evidence
Western blot images showing PADI4 band
PA2043 · PADI4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PADI4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PADI4 in Western blot, you can use spleen tissue lysate.
Positive control: Spleen (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: Spleen lysate is feasible, but signal may vary with eosinophil and neutrophil content.
HPA tissue expression evidence for PADI4
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 |
|---|---|---|---|---|
| Spleen | cells in red pulp | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PADI4 Western Blot Tips
Deeper troubleshooting and optimisation questions for PADI4, answered from its protein features.
Where should the main PADI4 band appear?
Band shift · The supplied predicted mass is 74.1 kDa for the 663-residue sequence. Use that as a reference, but assess any apparent mass difference against a molecular-weight marker; no observed band position is supplied.
Do the supplied features support multiple PADI4 isoform bands?
Isoforms · One isoform is listed, with no alternative sequence. The supplied features therefore do not identify an isoform that would account for an additional band.
Which PADI4 modification sites matter when choosing an antibody?
PTM · UniProt lists citrulline at positions 205, 212, 218, 372, 374, and 383. Check whether the antibody’s stated epitope overlaps these positions. These are UniProt coordinates; paper or antibody numbering may differ.
Does this guide establish induction of PADI4?
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 PADI4?
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 PA2043 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 PADI4 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Do the listed citrulline sites predict a visible band shift?
Interpretation · No. The six listed sites establish modification positions, but their presence alone does not establish a detectable shift or explain an apparent mass difference. Compare bands using matched samples before assigning a shift to citrullination.
PADI4 is listed in the cytoplasm and nucleus, and in cytoplasmic granules of eosinophils and neutrophils. If a fraction gives a weak signal, consider whether the preparation retains the compartment being examined.
Because PADI4 is reported in both cytoplasm and nucleus, quantify each fraction with a suitable fraction-specific loading reference. A change in one fraction alone need not represent a change in total PADI4.
Compare it with the 74.1 kDa prediction and verify antibody specificity. The supplied record gives no observed band position or alternative sequence, and its listed citrulline sites alone do not identify the cause of an extra band.
PADI4 is listed in the cytoplasm and nucleus, and in cytoplasmic granules of eosinophils and neutrophils. If a fraction gives a weak signal, consider whether the preparation retains the compartment being examined.
Because PADI4 is reported in both cytoplasm and nucleus, quantify each fraction with a suitable fraction-specific loading reference. A change in one fraction alone need not represent a change in total PADI4.
Compare it with the 74.1 kDa prediction and verify antibody specificity. The supplied record gives no observed band position or alternative sequence, and its listed citrulline sites alone do not identify the cause of an extra band.
Boster reagents
PADI4 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-PADI4/PAD4 Antibody Picoband®
Cat # PA2043
PA2043 is an anti-PADI4/PAD4 antibody listed for human reactivity. Its Western blot image shows PANC and 293T cell lysates; the supplied evidence does not establish performance in other samples.
Which to pick: PA2043 is the only listed option. Choose it for a human PADI4 Western blot when its PANC and 293T lysate image is relevant to your experiment.
Source: BosterBio PADI4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UM07.
- Human Protein Atlas. PADI4 tissue expression.
- PMC10657297 — target-verified WB comparison
- PMC3119849 — target-verified WB comparison
- PMC5308718 — target-verified WB comparison