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HPGD / 15-hydroxyprostaglandin dehydrogenase [NAD(+)] · Western blot design guide
Design a Western Blot for HPGD
Source-linked HPGD Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HPGD 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
HPGD 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 | ~29 kDa | |
| Observed band | ~26 kDa | |
| Gel | 12% (catalog A02686-3) | |
| Positive control | Duodenum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Below predicted mass | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked HPGD Western Blot Protocol Options
The A02686-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human RT4, human A549 (catalog A02686-3) |
| Gel % | 12% (catalog A02686-3) |
| Load | 30 ug; reducing conditions (catalog A02686-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02686-3) |
| Membrane | nitrocellulose membrane (catalog A02686-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02686-3) |
| Primary antibody | A02686-3 · 0.5 μg/mL (catalog A02686-3) |
| Primary incubation | overnight at 4°C (catalog A02686-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02686-3) |
| Secondary incubation | 1.5 hour at RT (catalog A02686-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02686-3) |
| Detection | ECL (catalog A02686-3) |
Section 2
What Is the Expected HPGD Western Blot Band Size?
HPGD is predicted at 29 kDa and observed at ~26 kDa in reducing whole-cell blots; the cause of the difference is not established.
| Band at ~26 kDa | Empirical HPGD band in reducing whole-cell lysates; confirm identity with a positive control |
| Band near 29 kDa | Near the UniProt predicted monomer mass; identity requires confirmation |
| Band near twice the monomer size | Could reflect a homodimer if it remains intact during electrophoresis |
| Several discrete bands | Five isoforms are listed, but distinct migration has not been established |
💡Expected HPGD appearanceHPGD has a predicted monomer mass of 29 kDa, while reducing whole-cell blots show ~26 kDa; the difference is unexplained, so confirm band identity with appropriate controls.
| UniProt predicted monomer mass | Predicts a band near 29 kDa; the observed band is ~26 kDa |
| Homodimer formation | Could produce a band near twice the monomer size if the dimer survives electrophoresis |
| Isoform 1 | Its individual apparent size is not supplied |
| Isoform 2 | May differ in size, but its individual apparent size is not supplied |
| Isoform 3 | May differ in size, but its individual apparent size is not supplied |
| Isoform 4 | May differ in size, but its individual apparent size is not supplied |
| Isoform 5 | May differ in size, but its individual apparent size is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | HPGD may be below detection in the tested cytoplasmic lysate | Check loading and assay performance with an HPGD-positive whole-cell lysate |
| Band higher than expected | An HPGD homodimer may persist during electrophoresis | Compare reducing and more thoroughly denatured samples; confirm identity by HPGD knockdown |
| Band lower than expected | The empirical ~26 kDa band runs below the 29 kDa prediction for an unestablished reason | Compare with a validated ~26 kDa positive-control band and HPGD knockdown |
| Multiple bands | Five HPGD isoforms are listed, but their migration is unverified | Check which bands respond to HPGD knockdown or an independent antibody |
| Weak or no signal | HPGD abundance in the tested lysate may be low | Verify transfer and loading, then compare with an HPGD-positive lysate |
Experiment evidence
Western blot images showing HPGD band
A02686-3 · HPGD
M02686 · HPGD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HPGD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HPGD in Western blot, you can use duodenum tissue, which has high HPA expression.
Positive control: Duodenum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside.
⚠️Feasibility: Cytoplasmic HPGD is suitable for tissue lysates; verify that adipose tissue gives no detectable band.
HPA tissue expression evidence for HPGD
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 |
|---|---|---|---|---|
| Duodenum | paneth cells | High | Protein (IHC) | HPA → |
| Lung | endothelial cells | High | Protein (IHC) | HPA → |
| Skin | fibrohistiocytic cells | High | Protein (IHC) | HPA → |
| Small intestine | paneth cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HPGD Western Blot Tips
Deeper troubleshooting and optimisation questions for HPGD, answered from its protein features.
How should HPGD 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 HPGD isoforms produce different bands?
Isoforms · UniProt lists five isoforms with distinct sequences. Isoform 3 lacks residues 1–121; isoform 5 lacks 73–140; isoform 4 changes 140–143 and lacks 144–266; isoform 2 replaces 167–266 with a shorter sequence. These differences could affect apparent bands, but the features alone do not establish which isoforms are expressed or visible.
Check its epitope against each listed isoform sequence. Residues 1–121 are absent from isoform 3, 73–140 from isoform 5, and 144–266 from isoform 4. Isoform 2 replaces residues 167–266. These are UniProt coordinates; antibody documentation may use another numbering convention.
Check its epitope against each listed isoform sequence. Residues 1–121 are absent from isoform 3, 73–140 from isoform 5, and 144–266 from isoform 4. Isoform 2 replaces residues 167–266. These are UniProt coordinates; antibody documentation may use another numbering convention.
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 HPGD?
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 HPGD?
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 A02686-3 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 HPGD bands be quantified across samples?
Quantitation · Use the same band definition across samples and verify that the measured signal corresponds to the intended HPGD isoform or isoforms. Five isoforms are listed, several with substantial sequence differences, so an antibody may recognize them differently. The supplied features do not establish that every HPGD band represents the same protein amount.
Why might HPGD appear near 26 kDa instead of 29 kDa?
Interpretation · The supplied observed band is approximately 26 kDa, while the predicted mass is 29 kDa. These values differ, but the listed features do not establish why. Compare the band with an appropriate molecular weight marker and confirm its identity before assigning a cause.
UniProt describes HPGD as a homodimer. That makes a higher band worth investigating, but homodimerization alone does not establish that a dimer survives sample preparation or appears on a Western blot. Check whether the band changes with preparation conditions before assigning it to a dimer.
UniProt places HPGD in the cytoplasm. Include a cytoplasmic fraction or whole-cell lysate when assessing its expression, and interpret a weak signal in another isolated fraction in light of that location.
UniProt describes HPGD as a homodimer. That makes a higher band worth investigating, but homodimerization alone does not establish that a dimer survives sample preparation or appears on a Western blot. Check whether the band changes with preparation conditions before assigning it to a dimer.
UniProt places HPGD in the cytoplasm. Include a cytoplasmic fraction or whole-cell lysate when assessing its expression, and interpret a weak signal in another isolated fraction in light of that location.
Boster reagents
HPGD 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-HPGD Antibody Picoband®
Cat # A02686-3
Real WB data
Anti-PGDH Rabbit Monoclonal Antibody
Cat # M02686
Both listed HPGD antibodies have WB images. A02686-3 shows human RT4 and A549 lysates, with a reported band near 26 kDa versus 29 kDa expected. M02686 shows SW480 lysate. The supplied evidence is limited to these examples.
Which to pick: For mouse or rat samples, A02686-3 lists reactivity, but its WB image uses human lysates. For human samples, both list reactivity and have WB images; A02686-3 provides detailed conditions, while M02686 has a brief SW480 caption.
Source: BosterBio HPGD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P15428.
- Human Protein Atlas. HPGD tissue expression.
- PMC11899648 — target-verified WB comparison
- PMC12288446 — target-verified WB comparison
- PMC7116369 — target-verified WB comparison