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PXDN / Peroxidasin homolog · IHC design guide
Design Immunohistochemistry for PXDN
Plan PXDN staining in paraffin sections around fibroblast-associated and extracellular positivity (HPA tissue IHC). This guide covers fixation, antibody dilution and chromogenic detection using a documented 1 μg/ml IHC-P workflow (datasheet: DZ33984).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PXDN Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Extracellular tissue staining (HPA tissue IHC) | |
| Staining pattern | Fibroblast-associated and extracellular positivity (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet DZ33984) | |
| Positive control | Bronchus+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Only processed PXDN integrates into the matrix (UniProt) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | 2 isoforms; a shorter form may lack the signal peptide, so check the epitope (UniProt) |
Section 1
Recommended PXDN IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet DZ33984) with three published PXDN IHC protocols (PMC11603849; PMC6627806; PMC8858314).
| Sample | Paraffin-embedded human pancrease cancer tissue; fixative not specified (datasheet DZ33984) |
| Fixation | Image fixative and duration unreported (datasheet DZ33984); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet DZ33984); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet DZ33984) |
| Primary antibody | Rabbit anti-PXDN, 1 μg/ml (datasheet DZ33984) |
| Primary incubation | Overnight at 4 °C (datasheet DZ33984) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet DZ33984) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PXDN-positive staining in ciliated cells (cilia axoneme) of bronchus (HPA tissue IHC: Medium). HPA tissue profile: Selective expression in fibroblasts and extra cellular positivity. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet DZ33984); adjust only if tissue staining or controls warrant it.
Section 2
What Is the Expected PXDN Staining Pattern?
PXDN is secreted into extracellular space and matrix, including basement membrane, and can also localise to the endoplasmic reticulum and cell surface; it has no transmembrane segment (UniProt Q92626). In paraffin-section IHC, expect selective fibroblast and extracellular staining, with medium staining reported in bronchial cilia axonemes and fallopian-tube cilia tips (HPA tissue IHC). HPA rates the tissue staining profile Approved (HPA tissue IHC).
| Selective staining around fibroblasts or in extracellular structures, with medium signal along bronchial cilia axonemes or at fallopian-tube cilia tips (HPA tissue IHC). | This matches the reported tissue profile and the ability of secreted PXDN to occupy extracellular matrix and basement membrane (HPA tissue IHC; UniProt Q92626). Assess the structure and its neighboring cells; extracellular signal need not trace a complete cell outline (UniProt Q92626 secretion). |
| Strong nuclear staining dominates, while extracellular structures and the reported ciliary sites are unstained. | A predominantly nuclear pattern is discordant with the listed secreted, matrix, cell-surface and endoplasmic-reticulum locations (UniProt Q92626). Treat it as suspect staining; compare the negative control and an HPA-reported positive structure before assigning it to PXDN (HPA tissue IHC). |
| Strong signal appears in adipocytes or adrenal glandular cells, despite little signal in the reported positive structures. | HPA reports PXDN as not detected in those specific cell populations, rather than declaring their whole tissues negative (HPA tissue IHC). Check for cross-reactivity or endogenous chromogenic detection activity, especially if the control without primary antibody also stains (standard IHC practice). |
| Chromogen coats most of the section, including stroma and cells, without a selective pattern. | Uniform colour obscures the selective fibroblast and extracellular profile reported by HPA (HPA tissue IHC). Review primary-antibody concentration, wash stringency and detection controls as general IHC background checks; the supplied sources do not establish a PXDN-specific cause for diffuse staining (standard IHC practice). |
| No convincing signal appears in bronchial cilia axonemes or fallopian-tube cilia tips. | Those structures have medium staining in HPA's tissue record, so an absent signal warrants a run-level check (HPA tissue IHC). Confirm that the sampled section contains the relevant ciliated cells, then review antibody, retrieval and detection conditions; a negative result alone does not prove PXDN is absent (standard IHC practice). |
💡Expected PXDN appearanceCall a result positive when staining is selective in fibroblast-associated or extracellular structures, or reaches medium intensity in bronchial cilia axonemes or fallopian-tube cilia tips (HPA tissue IHC); dominant nuclear or indiscriminate whole-section colour is suspect against that profile (UniProt Q92626; HPA tissue IHC).
| Secreted protein and compartment | PXDN occupies extracellular space, matrix and basement membrane, but is also listed at the cell surface and in the endoplasmic reticulum (UniProt Q92626). Score extracellular staining as well as cell-associated signal; tissue RNA and protein locations can differ for a secreted variant (HPA tissue IHC). |
| Proteolytic processing | Only proteolytically processed PXDN integrates into extracellular matrix (UniProt Q92626). A tissue section may therefore show a different balance of cellular and matrix staining; the supplied antibody record does not specify which processed forms its epitope detects (UniProt Q92626; HPA antibody record). |
| Isoforms and signal sequence | UniProt lists 2 isoforms and describes a shorter form, probably lacking the signal sequence, in testis and EB1 cells undergoing p53-dependent apoptosis (UniProt Q92626). Do not require every PXDN-positive context to show identical secretion; the supplied antibody record gives no isoform-specific staining validation (HPA antibody record). |
| Antibody validation | The supplied HPA antibody, HPA012375, is rated Approved for IHC; its ICC status is unreported, and no ICC-IF image cell lines are listed (HPA antibody record; HPA subcellular). The HPA subcellular summary says membrane and secreted, but supplies no main ICC-IF location, so it does not define an IF pattern (HPA subcellular). |
| Retrieval and detection settings | Antigen retrieval and primary-antibody concentration are routine paraffin-section IHC variables (standard IHC practice). Neither UniProt nor the supplied HPA record gives a PXDN-specific retrieval condition or fixation sensitivity; optimize these settings with controls and avoid treating a retrieval-dependent change as established PXDN biology (UniProt Q92626; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported ciliary or extracellular positives are blank. | The relevant structures may be absent from the section, or the IHC run may have insufficient usable signal; HPA reports medium staining at the specified ciliary sites (HPA tissue IHC). | Confirm ciliated cells are present and correctly oriented, then check the positive control, retrieval, primary-antibody incubation and detection steps in sequence (standard IHC practice). |
| Only a sharp nuclear pattern is visible. | Nuclear dominance conflicts with the supplied PXDN location record (UniProt Q92626). | Review the control without primary antibody and antibody specificity; require a reproducible, anatomically plausible extracellular or reported ciliary pattern before interpreting the signal as PXDN (HPA tissue IHC; standard IHC practice). |
| Adipocytes or adrenal glandular cells stain strongly. | Those cell types are listed as not detected by HPA; cross-reactivity or endogenous chromogenic activity is possible, but the pattern alone cannot distinguish them (HPA tissue IHC; standard IHC practice). | Compare a control without primary antibody and an HPA-reported positive structure in the same run; review blocking and detection if the control also develops colour (HPA tissue IHC; standard IHC practice). |
| Brown colour spreads across the section and masks tissue structure. | Excess antibody or incomplete removal of detection reagents can raise nonspecific background (standard IHC practice). | Review the antibody concentration, washes, peroxidase blocking and detection time against controls; score PXDN only where selective staining remains interpretable (standard IHC practice; HPA tissue IHC). |
| Extracellular signal is called negative because nearby cell bodies are pale. | That reading can miss secreted, matrix-associated PXDN; processed PXDN is the form that integrates into matrix (UniProt Q92626). | Inspect extracellular and basement-membrane regions alongside cell-associated staining, and record the compartment separately from the stained cell population (UniProt Q92626; standard IHC practice). |
| An IF image is used to set the paraffin-section IHC acceptance pattern. | The supplied HPA subcellular record has no ICC-IF image cell lines or main ICC-IF location, while the supplied antibody has an IHC Approved rating and no ICC rating (HPA subcellular; HPA antibody record). | Use the HPA tissue IHC observations to assess the chromogenic section. Treat IF localisation as a separate application requiring its own validation (HPA tissue IHC; HPA antibody record). |
Experiment evidence
IHC & IF images showing PXDN staining
DZ33984 · IHCIHC analysis of Peroxidasin homolog isoform X1 using anti-Peroxidasin homolog isoform X1 antibody (DZ33984). Peroxidasin homolog isoform X1
DZ33984 · IHCIHC analysis of Peroxidasin homolog isoform X1 using anti-Peroxidasin homolog isoform X1 antibody (DZ33984). Peroxidasin homolog isoform X1
A06456-1 · IHCIHC analysis of PXDN using anti-PXDN antibody (A06456-1). PXDN was detected in a paraffin-embedded section of human renal cancer tissue. Hea
A06456-1 · IHCIHC analysis of PXDN using anti-PXDN antibody (A06456-1). PXDN was detected in a paraffin-embedded section of human clear cell renal cell ca
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130508-PXDN/tissue/cerebral+cortex#imid_3306088
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130508-PXDN/tissue/colon#imid_3305998
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130508-PXDN/tissue/liver#imid_3306015
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130508-PXDN/tissue/kidney#imid_3306016
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PXDN IHC & IF
🧪Run bronchus first and assess staining in ciliated-cell cilia axonemes (HPA: Bronchus, ciliated cells [cilia axoneme], Medium). Run adipose tissue as the negative tissue (HPA: Adipose tissue, adipocytes, Not detected); on the bronchus slide, non-ciliated cells should lack the ciliary-axoneme pattern and can serve as a background comparator (HPA: bronchus signal assigned to ciliated-cell cilia axonemes).
Positive control tissue: Bronchus (Ciliated cells (cilia axoneme), HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PXDN; derive a cell-line control from the positive tissue's cell type (Ciliated cells (cilia axoneme)) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG controls, plus a PXDN-knockout specimen or validated immunizing-peptide competition as a biological specificity control (caption: rabbit primary antibody; standard IHC practice). For the biotin-based DAB workflow, check endogenous peroxidase and biotin background, especially near the ciliated surface (caption: biotinylated secondary, streptavidin-biotin complex and DAB; HPA: bronchus cilia-axoneme staining; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected DZ33984 paraffin-section caption does not report a fixative (caption: fixative unreported). The caption demonstrates staining after heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required; it provides no basis to call frozen sections or IF easier (caption: paraffin-section IHC with EDTA retrieval). Interpret staining at the ciliated surface cautiously because the HPA bronchus signal is assigned to cilia axonemes, while UniProt also places PXDN in extracellular matrix and basement membrane (HPA: Bronchus, ciliated cells [cilia axoneme], Medium; UniProt Q92626 subcellular location).
HPA tissue IHC evidence for PXDN
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Antibody staining in cells/structures not annotated, view images.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | Ciliated cells (cilia axoneme) | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (tip of cilia) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PXDN IHC Tips
Troubleshoot PXDN staining in paraffin sections by checking retrieval, compartment, background, and the scoring unit against the supplied tissue evidence.
Which retrieval condition should I try first when PXDN staining is weak?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet DZ33984). The selected tissue image used this condition, followed by 10% goat serum blocking and 1 µg/ml primary antibody overnight at 4°C (caption DZ33984). If staining remains weak, compare a different retrieval buffer on matched sections while keeping detection and exposure conditions constant (standard IHC practice). Judge improvement by whether the expected extracellular or fibroblast-associated pattern becomes clearer without a parallel rise in diffuse background; neither a stronger signal nor a different buffer alone establishes specificity (UniProt Q92626 localisation; HPA tissue IHC profile; standard IHC practice).
Could fixation explain weak or uneven PXDN staining?
The selected paraffin-section caption does not state a fixative, so PXDN-specific sensitivity to fixation is unknown for this antibody (caption DZ33984). Record the specimen’s actual fixative and processing history, then compare sections processed alike before attributing a weak result to fixation (standard IHC practice). Keep retrieval at EDTA pH 8.0 and primary antibody at the caption’s 1 µg/ml while assessing that comparison (datasheet DZ33984; caption DZ33984). If signal differs between processing batches, check section integrity and background alongside PXDN staining; the observed difference remains a processing association until tested with appropriately matched specimens (standard IHC practice).
Where should a convincing PXDN signal appear in paraffin sections?
Look for extracellular or basement-membrane-associated staining and evaluate fibroblast-associated positivity in its tissue context (UniProt Q92626 localisation; HPA tissue IHC profile). PXDN is secreted and can occur at the cell surface and in the endoplasmic reticulum, so a single cellular compartment is not an absolute requirement (UniProt Q92626 localisation). Its cell-surface staining may form hot spots, while extracellular-matrix incorporation is reported for proteolytically processed PXDN (UniProt Q92626, PubMed:25708780; PubMed:34679700). Compare the apparent compartment with tissue structure on the counterstained section and a matched negative control; widespread nuclear colour or staining confined to damaged edges warrants investigation before scoring (standard IHC practice).
Can this stain distinguish PXDN isoforms or its processed extracellular form?
PXDN has 2 reported isoforms and a precursor spanning residues 1–1479; an active fragment spans 27–1336 (UniProt Q92626 isoforms and processing). The selected image names isoform X1, but its caption does not locate the antibody epitope or demonstrate discrimination between forms (caption DZ33984). PXDN also has 10 listed glycosylation sites, making epitope accessibility a consideration without establishing that glycosylation changes this antibody’s staining (UniProt Q92626 glycosylation; standard IHC practice). Check the antibody’s documented epitope and validation before assigning isoform or cleavage status; report the observed IHC distribution as PXDN immunoreactivity when that information is unavailable (standard IHC practice).
How should I assess PXDN localisation in a multiplex IF experiment?
Pair PXDN with a marker for the expected cell population, such as a validated fibroblast marker, and inspect whether extracellular signal lies beside rather than inside marked cells (HPA tissue IHC profile; UniProt Q92626 localisation; standard IF practice). Include single-colour and unstained controls, and choose spectrally separated fluorophores after checking tissue autofluorescence in the intended channels (standard IF practice). For a documented intracellular epitope, compare controlled permeabilisation conditions; for an accessible extracellular or cell-surface epitope, assess staining without permeabilisation first (UniProt Q92626 topology and localisation; standard IF practice). The paraffin IHC caption supplies EDTA pH 8.0 retrieval evidence for DZ33984, not an IF fixation or permeabilisation condition (caption DZ33984).
How can I reduce diffuse DAB staining without losing PXDN signal?
Run a no-primary control and check endogenous peroxidase blocking before interpreting diffuse DAB colour (standard chromogenic IHC practice). The selected workflow used 10% goat serum blocking, biotinylated secondary antibody, a streptavidin–biotin complex, and DAB; assess background contributed by those steps with suitable controls (caption DZ33984; standard IHC practice). Compare matched sections while changing one blocking, washing, or detection variable at a time, keeping EDTA pH 8.0 retrieval as the reference condition (datasheet DZ33984; standard IHC practice). Retain genuine-looking extracellular staining only if it remains spatially coherent and exceeds control staining; PXDN’s secreted location alone does not validate diffuse colour (UniProt Q92626 localisation; standard IHC practice).
What should I measure when PXDN staining spans cells and matrix? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scoring compartment before analysis: use an H-score or percentage of positive cells for a specified cell population, and positive stained area per mm² for extracellular regions (standard IHC quantification practice; UniProt Q92626 localisation). Normalise cell scores to eligible cells and matrix area to the annotated tissue area, excluding folds, necrosis, and missing tissue by the same rules across slides (standard IHC quantification practice). Keep retrieval, DAB development, counterstain, and image acquisition consistent across the comparison (standard IHC practice). Report cellular and extracellular results separately, since PXDN secretion can separate the site of protein production from the site of staining (UniProt Q92626 localisation; HPA tissue IHC reliability description).
Which PXDN patterns suggest a true positive rather than an artefact?
Give greatest weight to staining that follows extracellular structures or a plausible cell-associated pattern and is stronger than matched negative controls (UniProt Q92626 localisation; HPA tissue IHC profile; standard IHC practice). HPA reports selective fibroblast and extracellular positivity, plus medium signal at ciliary structures in bronchus and fallopian tube, so evaluate the specific tissue and structure rather than imposing one universal pattern (HPA tissue IHC). Investigate staining restricted to section edges, necrosis, or the no-primary control, and check endogenous peroxidase before calling DAB signal PXDN (standard IHC practice). Apparent nuclear-only staining deserves independent validation because the supplied PXDN localisation record does not place it in the nucleus (UniProt Q92626 localisation).
Boster reagents
Best PXDN / Peroxidasin homolog IHC Antibodies
Two anti-PXDN antibodies have IHC images from human paraffin sections of renal or pancreatic cancer tissue; neither has IF images (catalog image captions; catalog IF image lists).
Real IHC data
Anti-human peroxidasin homolog isoform X1 Antibody
Cat # DZ33984
Real IHC data
Anti-PXDN Antibody ®
Cat # A06456-1
DZ33984 has IHC images from human pancreatic and renal cancer paraffin sections and lists Human, Mouse and Rat reactivity (DZ33984 image captions; catalog reactivity). A06456-1 has IHC images from human renal cancer paraffin sections and lists Human reactivity (A06456-1 image captions; catalog reactivity).
Which to pick: For human tissue IHC, choose either rabbit polyclonal antibody: DZ33984 was used at 1 μg/ml with EDTA pH 8 retrieval and biotin-based DAB detection, while A06456-1 was used at 2 μg/ml with EDTA pH 8 retrieval and peroxidase-based DAB detection (catalog clonality; each SKU’s IHC image captions). Neither SKU lists IF/ICC or provides an IF image, so there is no documented IF/ICC choice here (catalog applications; catalog IF image lists). DZ33984 is the cross-species candidate because its catalog lists Human, Mouse and Rat reactivity, but its shown IHC sections are human; both captions describe paraffin sections and leave the fixative unreported (catalog reactivity; each SKU’s IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92626 (PXDN_HUMAN, Peroxidasin homolog).
- Human Protein Atlas. PXDN tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PXDN subcellular location (ICC-IF): Membrane, Secreted.
- Human Protein Atlas. PXDN antibody validation summary (1 antibodies).
- The oncogenic role and prognostic value of PXDN in human stomach adenocarcinoma. BMC cancer 2024 — PMC11603849.
- Proteomics-Metabolomics Combined Approach Identifies Peroxidasin as a Protector against Metabolic and Oxidative Stress in Prostate Cancer. International journal of molecular sciences 2019 — PMC6627806.
- PXDN regulated by WTAP/YTHDF1-mediated m(6)A modification activates PI3K/AKT signaling pathway through extracellular matrix remodeling to promote progression in nasopharyngeal carcinoma. Journal of experimental & clinical cancer research : CR 2025 — PMC12825198.
- Peroxidasin Enhances Basal Phenotype and Inhibits Branching Morphogenesis in Breast Epithelial Progenitor Cell Line D492. Journal of mammary gland biology and neoplasia 2021 — PMC8858314.
- PubMed PMID:10441517 — UniProt-cited evidence.
- PubMed PMID:11103812 — UniProt-cited evidence.
- PubMed PMID:18929642 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PXDN (Peroxidasin homolog). Expected localisation Extracellular tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet DZ33984). Validated IHC antibody: DZ33984 at 1 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (cilia axoneme)); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human pancrease cancer tissue; fixative not specified (datasheet DZ33984); 2. Fixation: Image fixative and duration unreported (datasheet DZ33984); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet DZ33984); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet DZ33984); 8. Primary antibody: Rabbit anti-PXDN, 1 μg/ml (datasheet DZ33984); 9. Primary incubation: Overnight at 4 °C (datasheet DZ33984). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Only processed PXDN integrates into the matrix (UniProt); Isoform / epitope: 2 isoforms; a shorter form may lack the signal peptide, so check the epitope (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt Q92626, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Secreted, Datasheet DZ33984, PMC11603849, PMC6627806. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PXDN/immunohistochemistry-ihc-guide.