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HSD17B4 / Peroxisomal multifunctional enzyme type 2 · IHC design guide
Design Immunohistochemistry for HSD17B4
Plan HSD17B4 paraffin IHC around granular cytoplasmic staining, using hepatocytes as a high-staining reference (HPA tissue IHC). This guide covers fixation consistency, a 0.5–1 μg/mL antibody starting range (datasheet: A02923), chromogenic detection and cell-specific scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSD17B4 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 | Granular cytoplasm (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm across tissues; high in hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02923) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+3 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 | Lung macrophages can stain strongly (HPA tissue IHC) | |
| Regulation | Specific expression regulators not reported (UniProt) | |
| Isoform / epitope | 3 isoforms; processed chains warrant epitope mapping (UniProt) |
Section 1
Recommended HSD17B4 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A02923) and a published prostate cancer IHC method (PMC3607121) provide starting points.
| Sample | Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02923) |
| Fixation | Image fixative and duration unreported (datasheet A02923); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A02923) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02923) |
| Primary antibody | Rabbit anti-HSD17B4, 0.5-1μg/ml (datasheet A02923) |
| Primary incubation | Overnight at 4 °C (datasheet A02923) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02923) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSD17B4-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for the catalog antibody (datasheet A02923).
Section 2
What Is the Expected HSD17B4 Staining Pattern?
HSD17B4 is a peroxisomal protein with no transmembrane segment (UniProt P51659 topology). In paraffin-section IHC, expect granular cytoplasmic staining, particularly in liver hepatocytes, lung macrophages, and the listed gastrointestinal endocrine cells (HPA: granular cytoplasmic expression; High in those cells). HPA rates the tissue IHC profile Enhanced, citing consistency between antibody staining and RNA expression (HPA: Enhanced reliability).
| Discrete cytoplasmic granules in hepatocytes or lung macrophages. | This matches the reported tissue pattern and cellular distribution (HPA: granular cytoplasmic expression; High in hepatocytes and lung macrophages). The granules are consistent with peroxisomal localisation (UniProt P51659: peroxisome); IHC morphology alone does not identify individual organelles (general IHC practice). |
| Predominantly nuclear or cell-surface staining with little granular cytoplasmic signal. | Treat this as a localisation mismatch: HSD17B4 is peroxisomal and has no transmembrane segment (UniProt P51659 topology; HPA: granular cytoplasmic expression). Review the antibody and detection controls before assigning the signal to HSD17B4 (general IHC practice). |
| Strong staining in adipocytes, ovarian stroma cells, chondrocytes, or vaginal squamous epithelial cells. | These specific cell populations were reported as Not detected (HPA: tissue IHC). Check cell identity and controls; cross-reactivity or endogenous detection activity are possible explanations for unexpected chromogenic signal (general IHC practice). The HPA result does not classify every cell in those tissues as negative. |
| Uniform haze across cells, stroma, and section edges. | A diffuse deposit lacks the reported granular cytoplasmic pattern (HPA: tissue IHC profile). Excess primary antibody, insufficient washing, or endogenous detection activity can produce nonspecific chromogenic background (general IHC practice); a no-primary control helps separate detection background from primary-antibody staining. |
| No granular signal in hepatocytes on a liver section. | Liver hepatocytes are reported High (HPA: liver tissue IHC), so an absent signal calls for a run and specimen check before interpreting a test section as negative. HPA does not establish HSD17B4-specific fixation sensitivity or guarantee staining in every specimen. |
💡Expected HSD17B4 appearanceA convincing positive shows clear granular cytoplasmic staining in High-reporting cells such as hepatocytes or lung macrophages (HPA: tissue IHC); dominant nuclear, membrane, or uniform background colour is a pattern mismatch (UniProt P51659 topology; HPA: granular cytoplasmic expression).
| Tissue and cell selection | Liver hepatocytes, lung macrophages, and appendix, colon, duodenum, or rectum endocrine cells are reported High (HPA: tissue IHC). Choose a known-positive cell population within the section when judging a weak run; HPA staining levels describe those cells, not every cell in the tissue. |
| Compartment and topology | Peroxisomal localisation and the absence of a transmembrane segment support a granular cytoplasmic expectation (UniProt P51659 topology; HPA: tissue IHC and subcellular profile). A chromogenic granule pattern is compatible with that localisation, but does not by itself prove organelle identity (general IHC practice). |
| Antibody evidence | HPA021302, HPA021311, and HPA021479 each have Enhanced IHC status (HPA: antibody validation). That supports the reported pattern at the HPA level; it does not establish how an unspecified catalog antibody performs under different staining conditions. |
| Isoforms, processing, and epitope | UniProt lists three isoforms and chains spanning residues 1–736, 1–311, and 312–736 (UniProt P51659). The supplied record gives no antibody epitope, so it cannot predict which forms a particular reagent detects or explain a staining difference from chain location alone. |
| IF/ICC Q&A: where should signal appear? | Peroxisomal puncta are the expected IF/ICC localisation (HPA: Peroxisomes, enhanced; UniProt P51659: peroxisome). This supports the compartment interpretation of granular IHC, while IF/ICC observations remain a separate application and do not specify an IHC-P staining condition. |
| Antigen retrieval and fixation evidence | Retrieval conditions should be checked against the chosen antibody's IHC-P instructions and controlled empirically (general IHC practice). The supplied UniProt and HPA records give no target-specific fixation effect or retrieval setting; neither a tissue staining level nor a listed modification establishes one. |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver control has no hepatocyte signal. | A failed staining run, unsuitable control section, or ineffective antibody condition is possible (general IHC practice); hepatocytes are reported High (HPA: liver tissue IHC). | Confirm tissue and cell identity, verify the detection controls, and review the chosen antibody's IHC-P instructions before scoring experimental sections (general IHC practice). Do not infer an HSD17B4-specific fixation effect from this result. |
| Signal is mainly nuclear or outlines cell membranes. | That distribution conflicts with the granular cytoplasmic IHC profile and peroxisomal localisation (HPA: tissue IHC; UniProt P51659 topology). | Inspect the no-primary control and compare a known-positive cell population; reassess specificity if the compartment mismatch persists (general IHC practice). |
| Adipocytes or chondrocytes stain strongly. | HPA reports these cell types as Not detected (HPA: adipose tissue adipocytes; soft tissue chondrocytes). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm cell identity, examine the no-primary control, and seek concordance with an independently validated antibody if available (general IHC practice; HPA: multiple antibodies have Enhanced IHC status). |
| Brown colour is diffuse throughout the section. | Nonspecific primary-antibody binding or detection background can obscure granules (general IHC practice); diffuse colour is unlike the HPA profile (HPA: granular cytoplasmic expression). | Check a no-primary control, then review blocking, primary-antibody dilution, washing, and chromogen development under the chosen IHC-P workflow (general IHC practice). |
| A few weak dots appear in a reported High cell type. | Low contrast can make a granular pattern hard to distinguish from background (general IHC practice); HPA reports High staining in the specified positive cells (HPA: tissue IHC). | Compare the dots with background in adjacent cells and controls, then review the antibody's IHC-P conditions (general IHC practice). Do not score isolated dots as positive solely because the cell type is listed High. |
| Two antibodies give different cell or compartment patterns. | The records list Enhanced IHC status for three HPA antibodies but do not provide their epitopes or explain discordant staining (HPA: antibody validation; UniProt P51659: three isoforms and processed chains). | Compare the same cell populations and controls across runs; prioritise reproducible granular cytoplasmic staining in HPA-reported positive cells (HPA: tissue IHC; general IHC practice). Avoid attributing discordance to a particular isoform without epitope evidence. |
Experiment evidence
IHC & IF images showing HSD17B4 staining
A02923 · IHCIHC analysis of HSD17B4 using anti-HSD17B4 antibody (A02923). HSD17B4 was detected in paraffin-embedded section of human intestinal cancer t
A02923 · IHCIHC analysis of HSD17B4 using anti-HSD17B4 antibody (A02923). HSD17B4 was detected in paraffin-embedded section of human lung cancer tissues
PA1727 · IHCIHC analysis of HSD17B4 using anti-HSD17B4 antibody (PA1727). HSD17B4 was detected in a paraffin-embedded section of Human Lung Cancer tissu
PA1727 · IHCIHC analysis of HSD17B4 using anti-HSD17B4 antibody (PA1727). HSD17B4 was detected in a paraffin-embedded section of Human Intestinal Cancer
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000133835-HSD17B4/tissue/cerebral+cortex#imid_5776380
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000133835-HSD17B4/tissue/lung#imid_5774115
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000133835-HSD17B4/tissue/colon#imid_5774162
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000133835-HSD17B4/tissue/liver#imid_5774155
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021302; green = HSD17B4, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021302; green = HSD17B4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSD17B4 IHC & IF
🧪Run colon first and score its endocrine cells for staining (HPA: High in colon endocrine cells). Use adipose tissue adipocytes as the biological tissue negative (HPA: Not detected in adipocytes); on the colon slide, treat neighboring cells outside the endocrine compartment as a background comparison, without assuming they are HSD17B4-negative (HPA: High is specified for endocrine cells).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HSD17B4 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Peroxisomes (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit isotype control matched to the catalog primary, and HSD17B4-knockout material if available (selected-SKU caption: rabbit primary antibody). For colon IHC, block endogenous peroxidase and check endogenous biotin background because the caption uses biotinylated secondary antibody, streptavidin–biotin detection, and DAB (selected-SKU tissue-IHC caption).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A02923 paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). That caption reports citrate retrieval at pH 6 for 20 minutes, so use it as the starting retrieval condition for this IHC assay (selected-SKU tissue-IHC caption). IF/ICC offers a way to assess the expected peroxisomal localization (HPA: Peroxisomes, enhanced), but the supplied evidence does not establish that IF/ICC or frozen sections are easier; check colon background from the biotin-based detection system (selected-SKU tissue-IHC caption).
HPA tissue IHC evidence for HSD17B4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HSD17B4 IHC Tips
Troubleshoot HSD17B4 staining in paraffin sections by checking retrieval, signal localisation, controls and cell specific scoring before interpreting chromogenic signal.
What retrieval should I try first for weak HSD17B4 staining in paraffin sections?
Start with heat mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A02923). The selected paraffin section image used that treatment before incubation with the IHC validated antibody at 1 µg/mL overnight at 4°C (datasheet A02923). If staining is weak, first check section adhesion, heating consistency and adequate cooling, then compare a small retrieval time series on adjacent sections while holding detection constant (standard IHC practice). Judge improvement by stronger granular cytoplasmic signal in an expected positive cell population, such as hepatocytes, without a parallel rise in diffuse background (HPA: granular cytoplasmic expression; high in hepatocytes).
Could fixation explain weak or diffuse HSD17B4 staining?
The selected A02923 image identifies a paraffin embedded section but does not report its fixative, so target specific fixation sensitivity is unknown (datasheet A02923). Record the fixative, fixation duration and tissue processing history for each specimen before comparing staining intensity (standard IHC practice). If signal differs between batches, stain comparable sections together and vary one processing factor at a time, using identical citrate pH 6 retrieval for 20 minutes (datasheet A02923; standard IHC practice). Assess preserved morphology and granular cytoplasmic staining in expected cells; neither the reported peroxisomal localisation nor the listed modifications establish a preferred fixation condition (HPA: granular cytoplasmic expression; UniProt P51659 localisation and modified residues).
How should HSD17B4 signal look in a convincing IHC section?
Expect cytoplasmic granularity at light microscope resolution, consistent with the reported peroxisomal location, rather than a uniform nuclear pattern (HPA: granular cytoplasmic expression; HPA: peroxisomes; UniProt P51659 localisation). Inspect individual cells at high magnification and compare the distribution with adjacent negative controls processed through the same chromogenic workflow (standard IHC practice). HSD17B4 has no transmembrane segment, so a crisp plasma membrane outline alone is not a persuasive match to its annotated topology (UniProt P51659 topology). In mixed tissue, locate signal within identified cell types before calling the section positive; HPA reports high staining in liver hepatocytes and lung macrophages (HPA: high in hepatocytes; high in macrophages).
Can epitope choice change the HSD17B4 pattern I see?
HSD17B4 has 3 annotated isoforms and a full length 1–736 chain that can be processed into 1–311 and 312–736 chains (UniProt P51659 isoforms and processing). Map the antibody immunogen to those regions before attributing a staining difference to altered expression, because recognition depends on whether the epitope is present and accessible (UniProt P51659 processing; standard IHC practice). The record also lists modified residues, including phosphoserine at position 52, but does not establish an effect on this antibody’s tissue staining (UniProt P51659 modified residues). If epitope information is unavailable, describe staining as antibody dependent and compare with an independently validated epitope where practical (standard IHC practice).
How can IF help check an ambiguous chromogenic HSD17B4 pattern?
Use a separate IF optimisation on comparable material and multiplex HSD17B4 with a validated marker of the expected cell type, such as a macrophage marker when examining lung macrophages (HPA: high in lung macrophages; standard IF practice). Choose fluorophores after checking unstained tissue for autofluorescence, and include single stain controls to assess spectral overlap (standard IF practice). HSD17B4 is peroxisomal and has no transmembrane segment; permeabilise sufficiently to reach an intraperoxisomal epitope if the antibody targets one, while recognising that its epitope location is not supplied (UniProt P51659 localisation and topology; standard IF practice). Compare puncta with a validated peroxisome marker, using matched acquisition settings and controls before relating IF observations to the IHC pattern (HPA: peroxisomes; standard IF practice).
What should I check when the DAB section looks uniformly brown?
The selected image used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin biotin detection and DAB chromogen (datasheet A02923). Run a no primary control and examine whether brown signal persists in the same structures; also verify peroxidase quenching and secondary antibody specificity (standard IHC practice). Because this workflow uses biotin based detection, compare with an appropriate biotin control or alternative detection system if tissue wide signal remains (datasheet A02923; standard IHC practice). Optimise wash stringency and primary antibody concentration while preserving granular cytoplasmic signal in expected cells, rather than accepting diffuse staining as HSD17B4 expression (HPA: granular cytoplasmic expression; standard IHC practice).
How should I quantify HSD17B4 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis region before scoring, because HPA reports different staining levels across cell types, including high staining in hepatocytes and undetected staining in adipocytes (HPA: high in hepatocytes; not detected in adipocytes). For chromogenic sections, report an H score from staining intensity and percentage of positive cells, or report percentage positive cells alone when intensity grading is unreliable (standard IHC practice). If counting granular positive profiles, express density per mm² of viable tissue and state the threshold used (standard IHC practice). Normalise comparisons to the same eligible cell population, section area, staining batch and image analysis settings, with background controls defining the positivity threshold (standard IHC practice).
When is an apparent HSD17B4 positive cell likely an artefact?
Treat granular cytoplasmic staining in an expected cell type as more credible than isolated nuclear or membrane only signal, given the reported peroxisomal location and tissue pattern (HPA: peroxisomes; HPA: granular cytoplasmic expression; UniProt P51659 topology). Check whether staining follows tissue edges, damaged or necrotic regions, or a no primary control, which would support a technical explanation (standard IHC practice). Residual endogenous peroxidase can produce chromogenic signal, so confirm adequate blocking when DAB persists without primary antibody (standard IHC practice). Interpret unexpected positive cells cautiously and compare them with the documented cell specific pattern, including high lung macrophage staining and undetected adipocyte staining (HPA: high in lung macrophages; not detected in adipocytes).
Boster reagents
Best HSD17B4 / Peroxisomal multifunctional enzyme type 2 IHC Antibodies
Both anti-HSD17B4 antibodies have IHC images from paraffin sections of human cancer tissue (catalog image captions). Both list Human, Mouse, and Rat reactivity (catalog reactivity fields); neither has IF image data (catalog image alts).
Real IHC data
Anti-Hydroxysteroid (17-beta) Dehydrogenase 4/HSD17B4 Antibody ®
Cat # A02923
Real IHC data
Anti-Hydroxysteroid (17-beta) Dehydrogenase 4/HSD17B4 Antibody ®
Cat # PA1727
A02923 has IHC images from human intestinal, lung, and mammary cancer paraffin sections, and lists Human, Mouse, and Rat reactivity (A02923 image captions; catalog reactivity). PA1727 has IHC images from human lung and intestinal cancer paraffin sections, and lists Human, Mouse, and Rat reactivity (PA1727 image captions; catalog reactivity).
Which to pick: For tissue IHC, choose A02923 if using citrate retrieval at pH 6 with biotin-based DAB detection (A02923 image caption), or PA1727 if using EDTA retrieval at pH 8 with peroxidase-based DAB detection (PA1727 image caption); both captions describe paraffin sections but do not report the fixative (catalog image captions). For IF/ICC, neither SKU lists that application or provides an IF figure (catalog applications; catalog IF image alts). For cross-species IHC, PA1727 explicitly lists Human, Mouse, and Rat for paraffin-section IHC, while A02923's IHC dilution text specifies Human; the IHC figures show human tissue only, and clonality is unreported for both (catalog dilution_raw; catalog image captions; catalog clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51659 (DHB4_HUMAN, Peroxisomal multifunctional enzyme type 2).
- Human Protein Atlas. HSD17B4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HSD17B4 subcellular location (ICC-IF): Localized to the peroxisomes..
- Human Protein Atlas. HSD17B4 antibody validation summary (3 antibodies).
- Acetylation-mediated degradation of HSD17B4 regulates the progression of prostate cancer. Aging 2020 — PMC7425433.
- Androgen deprivation promotes intratumoral synthesis of dihydrotestosterone from androgen metabolites in prostate cancer. Scientific reports 2013 — PMC3607121.
- A prediction model for prognosis of gastric adenocarcinoma based on six metabolism-related genes. Biochemistry and biophysics reports 2023 — PMC9957706.
- Longitudinal Plasma Proteomics-Derived Biomarkers Predict Response to MET Inhibitors for MET-Dysregulated NSCLC. Cancers 2023 — PMC9818927.
- PubMed PMID:7487879 — UniProt-cited evidence.
- PubMed PMID:9089413 — UniProt-cited evidence.
- PubMed PMID:9880674 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSD17B4 (Peroxisomal multifunctional enzyme type 2). Expected localisation Granular cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02923). Validated IHC antibody: A02923 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02923); 2. Fixation: Image fixative and duration unreported (datasheet A02923); 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: Citrate pH 6 , 20 min (datasheet A02923); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02923); 8. Primary antibody: Rabbit anti-HSD17B4, 0.5-1μg/ml (datasheet A02923); 9. Primary incubation: Overnight at 4 °C (datasheet A02923). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Lung macrophages can stain strongly (HPA tissue IHC); Isoform / epitope: 3 isoforms; processed chains warrant epitope mapping (UniProt); Regulation: Specific expression regulators not reported (UniProt). Evidence: UniProt P51659, HPA tissue IHC · Enhanced, HPA ICC-IF · peroxisomes., Datasheet A02923, PMC3607121. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSD17B4/immunohistochemistry-ihc-guide.