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PRTN3 / Myeloblastin · IHC design guide
Design Immunohistochemistry for PRTN3
Plan chromogenic IHC for PRTN3 in paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet A03863-3). Score cytoplasmic staining in immune cell subsets, especially bone marrow hematopoietic cells and splenic red pulp cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRTN3 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 | Cytoplasmic in immune cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic immune cell subsets, most abundant in marrow and spleen (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03863-3) | |
| Positive control | Bone marrow+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 | Secreted PRTN3 can separate protein from RNA location (HPA tissue IHC) | |
| Regulation | TNF or LPS stimulation prompts secretion (UniProt) | |
| Isoform / epitope | No listed isoforms; map epitopes to mature chain 28–248 (UniProt) |
Section 1
Recommended PRTN3 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet A03863-3). The published IHC protocols below cover vulvar, gastric, and colorectal cancer specimens (PMC7793473; PMC11912076; PMC10835931).
| Sample | Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A03863-3) |
| Fixation | Image fixative and duration unreported (datasheet A03863-3); 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 A03863-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03863-3) |
| Primary antibody | Rabbit anti-PRTN3, 2-5 μg/ml (datasheet A03863-3) |
| Primary incubation | Overnight at 4 °C (datasheet A03863-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03863-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRTN3-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in subsets of immune cells in several tissues, most abundant in bone marrow and spleen. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A03863-3); use each published retrieval condition with its corresponding antibody protocol (PMC7793473; PMC10835931).
Section 2
What Is the Expected PRTN3 Staining Pattern?
PRTN3 staining should be concentrated in subsets of immune cells, especially hematopoietic cells in bone marrow and cells in splenic red pulp (HPA: High; tissue IHC reliability Enhanced). Its expected pattern is cytoplasmic, consistent with predominant localization to neutrophil azurophil granules (HPA: cytoplasmic tissue expression; UniProt P24158: cytoplasmic granule). PRTN3 has no transmembrane segment, although secreted protein can be tethered to the neutrophil surface (UniProt P24158: topology and CD177 interaction).
| Strong cytoplasmic staining in a subset of bone marrow hematopoietic cells, with staining also seen in splenic red pulp cells (HPA: High in both sites). | This fits the tissue IHC pattern; assess the stained cells individually rather than treating either whole tissue as uniformly positive (HPA: cytoplasmic expression in subsets of immune cells). Granular cytoplasmic staining is biologically plausible because PRTN3 predominates in neutrophil azurophil granules (UniProt P24158: subcellular location). |
| Predominantly nuclear staining, with little cytoplasmic staining, in otherwise plausible positive cells. | A nuclear-dominant pattern does not match the reported localization (HPA: cytoplasmic tissue expression; UniProt P24158: cytoplasmic granule). Treat it as an unconfirmed staining pattern and check controls and detection conditions before scoring those cells as PRTN3 positive. |
| Strong staining of adipocytes or respiratory epithelial cells across a section. | These are poor matches for the reported cell distribution: adipocytes in adipose tissue and respiratory epithelial cells in bronchus were not detected (HPA: tissue IHC). Consider cross-reactivity or endogenous chromogenic detection activity; confirm whether nearby immune cells account for any localized signal. |
| Color deposits spread across cells and surrounding tissue without a clear cellular pattern. | Diffuse background cannot establish PRTN3 expression. It obscures the subset-restricted cytoplasmic pattern used for interpretation (HPA: tissue IHC). Compare with detection controls and inspect whether deposits track tissue edges or broad areas rather than individual cells. |
| No staining in bone marrow hematopoietic cells while the section otherwise appears interpretable. | This conflicts with a reported high-staining population (HPA: High in bone marrow hematopoietic cells). First check whether that population is present on the tested section, then assess the antibody and detection workflow; a blank slide alone cannot establish absent PRTN3. |
💡Expected PRTN3 appearanceA convincing positive is high cytoplasmic staining in a subset of bone marrow hematopoietic cells or splenic red pulp cells (HPA: tissue IHC); uniform nuclear or broad epithelial staining is a poor match (HPA: cytoplasmic pattern; bronchial respiratory epithelial cells Not detected).
| Choice of IHC antibody | HPA005938 has IHC Enhanced validation; CAB017558 has IHC Supported validation (HPA: antibody records). HPA notes medium consistency between staining and RNA data and disregarded presumed off-target binding (HPA: reliability description), so use cell identity and compartment alongside stain intensity. |
| Granule storage and release | PRTN3 predominates in neutrophil azurophil granules and can be secreted after stimulation or during transmigration (UniProt P24158: subcellular location). Interpret focal extracellular or peripheral color cautiously; its presence does not by itself identify the producing cell. |
| Surface-associated signal | Secreted PRTN3 can bind CD177 at the neutrophil surface despite having no transmembrane segment (UniProt P24158: topology and CD177 interaction). A peripheral signal can therefore be plausible when cell identity and cytoplasmic staining also support the call. |
| Maturation and epitope interpretation | The annotated precursor has a signal peptide, propeptides and a mature myeloblastin chain (UniProt P24158: processing). No antibody epitope is supplied here, so this record cannot predict which processed form a given antibody recognizes. |
| IF/ICC Q: Should fluorescence be exclusively granular? | No. HPA reports approved cytosolic localization with additional vesicle localization in ICC-IF (HPA: subcellular summary); UniProt describes predominant neutrophil granule localization (UniProt P24158: subcellular location). Interpret fluorescence in its own cellular context rather than requiring every positive cell to show resolvable puncta. |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow positive control is blank. | The expected high-staining hematopoietic population may be absent from the examined area, or staining or detection may have failed (HPA: High in bone marrow hematopoietic cells). | Locate hematopoietic cells on the counterstained section, then review the IHC-validated antibody, antigen retrieval and detection steps as general workflow checks; do not infer a PRTN3-specific fixation effect. |
| The whole section has diffuse chromogenic haze. | Nonspecific background or endogenous detection activity may obscure the expected subset-restricted cytoplasmic signal (HPA: tissue IHC pattern). | Compare detection-only controls, review blocking and washing, and score only cells with a distinct interpretable pattern; these are general chromogenic IHC checks. |
| Many nuclei are strongly stained. | Nuclear-dominant signal conflicts with the reported cytoplasmic tissue pattern and granule location (HPA: tissue IHC; UniProt P24158: subcellular location). | Check a known-positive region, inspect counterstain and chromogen separately, and reconsider specificity before assigning nuclear staining to PRTN3. |
| Adipocytes or bronchial respiratory epithelial cells stain strongly. | Those cell types are listed as Not detected (HPA: adipose tissue and bronchus IHC); cross-reactivity or endogenous activity is possible. | Use appropriate detection controls and compare nearby immune cells; do not count a broadly stained negative cell type as confirmation of PRTN3. |
| Only splenic red pulp shows signal, with uncertain cell identity. | HPA reports high staining in red pulp cells but does not identify every stained cell as a neutrophil (HPA: spleen tissue IHC). | Use the counterstained morphology to identify individual cells and report the observed location conservatively; avoid assigning neutrophil identity from PRTN3 color alone. |
| Peripheral staining appears beside cytoplasmic signal. | Secreted PRTN3 can be tethered to neutrophil membranes by CD177 (UniProt P24158: secretion and CD177 interaction). | Check whether the signal outlines identifiable immune cells and whether a cytoplasmic subset remains visible; flag isolated extracellular deposits as uncertain rather than scoring them as cells. |
Experiment evidence
IHC & IF images showing PRTN3 staining
A03863-3 · IHCIHC analysis of PRTN3 using anti-PRTN3 antibody (A03863-3). PRTN3 was detected in a paraffin-embedded section of human lung adenocarcinoma t
A03863-3 · IHCIHC analysis of PRTN3 using anti-PRTN3 antibody (A03863-3). PRTN3 was detected in a paraffin-embedded section of human tonsil tissue. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196415-PRTN3/tissue/cerebral+cortex#imid_2344648
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196415-PRTN3/tissue/colon#imid_2327510
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196415-PRTN3/tissue/liver#imid_2327495
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000196415-PRTN3/tissue/kidney#imid_2327504
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005938; green = PRTN3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005938; green = PRTN3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRTN3 IHC & IF
🧪Run bone marrow first and require staining in hematopoietic cells (HPA: High in bone marrow hematopoietic cells). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); morphologically identified nonhematopoietic cells on the bone marrow slide should lack specific granular staining (UniProt P24158 tissue specificity and granule localisation).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRTN3 in A-431, THP-1, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected A03863-3 caption: rabbit primary antibody and anti-rabbit secondary); use PRTN3-knockout material as a biological negative where available. Block endogenous peroxidase and check for residual signal in bone marrow before interpreting DAB staining (selected A03863-3 caption: HRP/DAB detection; UniProt P24158: expression in neutrophils).
⚠️Feasibility: The selected A03863-3 caption shows paraffin-section staining after heat retrieval in EDTA at pH 8.0, but its fixative is unreported; the supplied evidence reports no target-specific fixation window or fixation effect. Retrieval conditions therefore have a documented starting point, while frozen sections and IF cannot be ranked as easier from the supplied evidence (selected A03863-3 tissue-IHC caption). In bone marrow, endogenous peroxidase can complicate chromogenic interpretation; assess the blocked no-primary control alongside the stained section (selected A03863-3 caption: HRP/DAB detection).
HPA tissue IHC evidence for PRTN3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Presumed off target binding observed and disregarded. 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PRTN3 IHC Tips
Troubleshoot chromogenic PRTN3 staining in paraffin sections using the catalog antibody’s tissue image and the expected distribution in immune cells.
Which retrieval conditions should I start with for PRTN3 in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A03863-3). The selected tissue image used this retrieval before staining with 2 μg/ml antibody overnight at 4°C (datasheet A03863-3). If staining is weak, compare a second retrieval condition on matched sections while holding antibody concentration and detection constant (standard IHC practice). Judge recovery in hematopoietic cells of bone marrow or red-pulp cells of spleen, where staining is reported as high (HPA tissue IHC). Monitor section integrity and staining of adjacent negative compartments so increased signal is not mistaken for improved specificity (standard IHC practice).
How should I assess fixation when PRTN3 staining is weak?
The selected PRTN3 image identifies a paraffin-embedded section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A03863-3). Record the fixative, fixation duration, tissue thickness and processing history for each section before comparing staining (standard IHC practice). Run sections processed under different documented conditions through the same EDTA pH 8.0 retrieval and antibody workflow (datasheet A03863-3; standard IHC practice). Include a hematopoietic-cell-rich control because bone marrow hematopoietic cells show high staining (HPA tissue IHC). Interpret any condition-dependent difference as an observation from that comparison, without assigning a PRTN3-specific fixation mechanism from tissue patterns or protein features alone (standard IHC practice).
Where should convincing PRTN3 chromogenic staining appear?
Assess cytoplasmic staining in the relevant immune-cell subset: tissue IHC reports cytoplasmic expression in subsets of immune cells, with abundant signal in bone marrow and spleen (HPA tissue IHC). PRTN3 predominantly occupies neutrophil azurophil granules, so a granular cytoplasmic pattern is biologically plausible (UniProt P24158 subcellular location). Secreted PRTN3 can become tethered to the cell surface through CD177 after neutrophil stimulation (UniProt P24158 subcellular location). Evaluate apparent surface or extracellular DAB alongside cell morphology and the local inflammatory context, since compartment alone cannot establish which pool was stained (UniProt P24158 subcellular location; standard IHC practice). Treat widespread staining of unrelated tissue cells as a specificity concern (HPA tissue IHC; standard IHC practice).
Could precursor processing or glycosylation change the PRTN3 epitope detected?
PRTN3 has no listed isoforms, while its 256-amino-acid precursor is processed to a 28–248 myeloblastin chain (UniProt P24158 processing and isoforms). The record also lists glycosylation sites at residues 129 and 174 (UniProt P24158 glycosylation). If the catalog antibody’s epitope is known, map it against the cleaved terminal regions and mature chain before interpreting a missing or altered signal (UniProt P24158 processing; standard IHC practice). The supplied tissue-image caption does not identify that epitope, so it cannot establish which processed form is detected (datasheet A03863-3). Compare staining across matched sections under fixed retrieval and detection conditions before attributing differences to processing or glycosylation (standard IHC practice).
How can IF help assess PRTN3 localisation seen by IHC?
Use IF on a separate, appropriately validated preparation to compare PRTN3 with a marker of the expected neutrophil or immune-cell population (UniProt P24158 tissue specificity; HPA tissue IHC; standard IF practice). HPA reports cytosol and vesicles in its subcellular IF data, while UniProt places PRTN3 predominantly in neutrophil azurophil granules (HPA subcellular; UniProt P24158 subcellular location). Choose fluorophores after checking tissue autofluorescence and include single-stain controls for multiplex bleed-through (standard IF practice). For an intracellular epitope, evaluate permeabilisation; for an accessible surface epitope, compare staining without permeabilisation, given CD177-dependent surface tethering (UniProt P24158 subcellular location; standard IF practice). Do not transfer the paraffin-section retrieval conditions to IF without validation (datasheet A03863-3; standard IF practice).
What should I check when PRTN3 DAB staining is diffuse?
Begin with a no-primary control and inspect whether brown signal persists in leukocyte-rich or damaged areas (standard IHC practice). Block endogenous peroxidase before HRP detection, then compare matched sections with and without primary antibody to separate detection background from antibody-associated staining (standard IHC practice). The selected image used 10% goat serum, 2 μg/ml primary antibody and an HRP/DAB detection workflow (datasheet A03863-3). Titrate primary antibody and detection development while keeping retrieval fixed, and examine section edges, folds and necrotic regions for nonspecific deposition (standard IHC practice). Interpret diffuse extracellular signal cautiously because stimulated neutrophils can secrete PRTN3 (UniProt P24158 subcellular location).
How should I quantify PRTN3-positive cells across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and eligible cell population before scoring, using morphology or a matched cell-type stain to distinguish immune cells from surrounding tissue (HPA tissue IHC; standard IHC practice). Report the percentage of PRTN3-positive eligible cells and, where recruitment matters, positive-cell density per mm² of viable tissue (standard IHC practice). An H-score from 0–300 can summarize intensity and positive-cell fraction when staining is assessable per cell (standard IHC practice). Normalize cell positivity to the number of eligible cells, and density to measured viable area; exclude folds, edges and necrosis consistently (standard IHC practice). Record cytoplasmic and convincing surface-associated patterns separately because PRTN3 can occupy both locations (UniProt P24158 subcellular location).
How do I distinguish true PRTN3 staining from artefact?
A credible result places PRTN3 signal in an appropriate immune-cell population, especially cytoplasmic staining of cells in bone marrow or splenic red pulp (HPA tissue IHC). Neutrophil granules and possible CD177-associated surface signal provide compartment clues, but neither pattern alone proves antibody specificity (UniProt P24158 subcellular location; standard IHC practice). Reconsider broad staining in unrelated cells, particularly where HPA reports no detection, and check section edges or necrosis for concentrated DAB deposits (HPA tissue IHC; standard IHC practice). Compare no-primary and peroxidase-block controls when brown deposits suggest endogenous enzyme activity (standard IHC practice). HPA rates tissue evidence Enhanced but notes presumed off-target binding, so assess unexpected staining against morphology and controls (HPA tissue IHC).
Boster reagents
Best PRTN3 / Myeloblastin IHC Antibodies
Two human-reactive anti-PRTN3 antibodies list IHC use (catalog applications/reactivity). A03863-3 has human paraffin-section IHC images and catalog-listed IF/ICC use (A03863-3 image captions; catalog applications).
Real IHC data
Anti-PRTN3 Antibody ®
Cat # A03863-3
A03863-3 has IHC images from paraffin sections of human lung adenocarcinoma and tonsil, and is listed for IF/ICC without a supplied IF image (A03863-3 image captions; catalog applications and IF image alts). M03863-2 is listed for human IHC, with no supplied IHC image (M03863-2 catalog applications/reactivity and IHC image alts).
Which to pick: For tissue IHC, choose A03863-3: its own captions document human paraffin sections, EDTA retrieval at pH 8.0, and 2 μg/ml primary antibody; the fixative is unreported (A03863-3 IHC image captions). For IF/ICC, choose A03863-3 at the catalog-listed 5 μg/ml; M03863-2 is a rabbit monoclonal listed for human IHC but has no supplied IHC image (A03863-3 catalog applications/dilution; M03863-2 catalog clone/applications/reactivity and IHC image alts). Neither SKU has listed cross-species reactivity (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P24158 (PRTN3_HUMAN, Myeloblastin).
- Human Protein Atlas. PRTN3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PRTN3 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to vesicles..
- Human Protein Atlas. PRTN3 antibody validation summary (2 antibodies).
- Inflammatory Proteins HMGA2 and PRTN3 as Drivers of Vulvar Squamous Cell Carcinoma Progression. Cancers 2020 — PMC7793473.
- Low expression of PRTN3 regulates the progression of gastric cancer by inhibition of cell cycle and promotion of apoptosis. Translational cancer research 2025 — PMC11912076.
- Research on the process and molecular mechanism of inhibiting serous ovarian cancer by PRTN3 and its inhibitor Sivelestat. Journal of ovarian research 2025 — PMC12482878.
- High serum proteinase-3 levels predict poor progression-free survival and lower efficacy of bevacizumab in metastatic colorectal cancer. BMC cancer 2024 — PMC10835931.
- PubMed PMID:1681549 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRTN3 (Myeloblastin). Expected localisation Cytoplasmic in immune cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03863-3). Validated IHC antibody: A03863-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung adenocarcinoma tissue; fixative not specified (datasheet A03863-3); 2. Fixation: Image fixative and duration unreported (datasheet A03863-3); 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 A03863-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03863-3); 8. Primary antibody: Rabbit anti-PRTN3, 2-5 μg/ml (datasheet A03863-3); 9. Primary incubation: Overnight at 4 °C (datasheet A03863-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secreted PRTN3 can separate protein from RNA location (HPA tissue IHC); Isoform / epitope: No listed isoforms; map epitopes to mature chain 28–248 (UniProt); Regulation: TNF or LPS stimulation prompts secretion (UniProt). Evidence: UniProt P24158, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to vesicles., Datasheet A03863-3, PMC7793473, PMC11912076. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRTN3/immunohistochemistry-ihc-guide.