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- Table of Contents
PPARD / Peroxisome proliferator-activated receptor delta · Western blot design guide
Design a Western Blot for PPARD
Real validated PPARD Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PPARD 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
PPARD Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~49.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Heart muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated PPARD Western Blot Protocols
The A01557 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 | A01557; 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 PPARD Western Blot Band Size?
PPARD's predicted mass is 49.9 kDa; four annotated isoforms could affect migration, but no empirical band size or distinct isoform bands are established.
| Single band near 49.9 kDa | Consistent with the predicted PPARD mass; confirm identity with controls |
| Several discrete bands | Could reflect isoforms 1, 2, 3, and 4; their migration is not established |
| One band despite four annotated isoforms | The isoforms need not resolve into separate bands |
| Weak band in a cytosolic fraction | Consistent with PPARD's nuclear location |
💡Expected PPARD appearancePPARD has a predicted mass of 49.9 kDa, but no empirical band size is supplied; use antibody specificity controls to confirm any band near that size.
| UniProt predicted mass | Places the expected band near 49.9 kDa |
| Splice isoform 1 | May migrate differently from other isoforms; its individual mass is not supplied |
| Splice isoform 2 | May migrate differently from other isoforms; its individual mass is not supplied |
| Splice isoforms 3 and 4 | May differ in migration; their individual masses and separation are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | A preparation depleted of nuclear proteins may contain little PPARD | Check a nuclear fraction and verify fraction quality |
| Band higher than expected | An isoform or unrelated antibody target may migrate above the predicted mass | Check antibody specificity by PPARD knockdown or knockout |
| Band lower than expected | An isoform or unrelated antibody target may migrate below the predicted mass | Check antibody specificity by PPARD knockdown or knockout |
| Multiple bands | Annotated isoforms or unrelated antibody targets may contribute | Compare bands after PPARD knockdown or knockout |
| Weak or no signal | PPARD is nuclear and may be underrepresented in the sampled fraction | Enrich nuclear protein and confirm extraction quality |
Experiment evidence
Western blot images showing PPARD band
A01557 · PPARD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PPARD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PPARD in Western blot, you can use adrenal gland tissue, which HPA scores High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Heart muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: PPARD is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for PPARD
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced PPARD Western Blot Tips
Deeper troubleshooting and optimisation questions for PPARD, answered from its protein features.
How should PPARD 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.
Could PPARD isoforms produce bands at different sizes?
Isoforms · Yes. UniProt lists four isoforms with sequence differences that could change calculated mass. Their presence alone does not establish which isoforms are expressed or whether separate bands will resolve.
Using canonical UniProt coordinates, isoform 4 lacks residues 44–141. Isoform 2 has DR replaced by GE at 360–361 and lacks residues 362–441. Check whether your antibody recognizes a region retained in the isoform you want to detect.
At canonical UniProt residues 2–43, isoform 3 replaces EQPQEEAPEVREEEEKEEVAEAEGAPELNGGPQHALPSSSYT with HQR. An antibody targeting that canonical N-terminal segment may miss isoform 3.
Using canonical UniProt coordinates, isoform 4 lacks residues 44–141. Isoform 2 has DR replaced by GE at 360–361 and lacks residues 362–441. Check whether your antibody recognizes a region retained in the isoform you want to detect.
At canonical UniProt residues 2–43, isoform 3 replaces EQPQEEAPEVREEEEKEEVAEAEGAPELNGGPQHALPSSSYT with HQR. An antibody targeting that canonical N-terminal segment may miss isoform 3.
Do UniProt features support a modification-dependent PPARD band shift?
PTM · The supplied record includes a Ubl conjugation keyword but lists no modified-residue coordinates. That keyword alone does not establish a visible shift or explain a difference from the predicted 49.9 kDa mass.
Does this guide establish induction of PPARD?
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 PPARD?
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 A01557 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.
What should I consider when quantifying PPARD bands?
Quantitation · PPARD is annotated as nuclear and has four isoforms. Keep sample preparation consistent, and establish which isoforms the antibody can detect before combining band intensities. Quantify only bands whose identity has been supported.
What molecular weight should I expect for PPARD?
Interpretation · The predicted mass is 49.9 kDa for the canonical 441-residue sequence. No empirical apparent band position is supplied, so use 49.9 kDa as a reference rather than an exact migration target.
Compare them with the predicted 49.9 kDa canonical protein and the listed isoform sequence changes. Confirm that the antibody target is retained in each candidate isoform. The supplied features cannot assign an unexpected band to a particular isoform or modification.
Compare them with the predicted 49.9 kDa canonical protein and the listed isoform sequence changes. Confirm that the antibody target is retained in each candidate isoform. The supplied features cannot assign an unexpected band to a particular isoform or modification.
Boster reagents
PPARD 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-PPARD Antibody (C-term)
Cat # A01557
A01557 is a human-reactive anti-PPARD antibody with a Western blot image from human placenta tissue lysate at 35 µg per lane. The supplied evidence shows this tested context only; it does not establish performance in other samples.
Which to pick: A01557 is the only listed PPARD antibody. Choose it for a human Western blot starting point; its documented image uses human placenta lysate at 35 µg per lane.
Source: BosterBio PPARD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q03181.
- Human Protein Atlas. PPARD tissue expression.
- PMC13359835 — target-verified WB comparison
- PMC9827666 — target-verified WB comparison
- PMC11772773 — target-verified WB comparison