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HSD3B7 / 3 beta-hydroxysteroid dehydrogenase type 7 · IHC design guide
Design Immunohistochemistry for HSD3B7
Plan HSD3B7 paraffin IHC around the reported cytoplasmic tissue staining (HPA tissue IHC) and ER membrane localization (UniProt). The catalog antibody has an IHC range of 2–5 μg/ml (datasheet A10025-1); include controls when interpreting staining because antibody staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSD3B7 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 tissue staining (HPA tissue IHC); ER membrane localization (UniProt) | |
| Staining pattern | Cytoplasmic staining in several tissue types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10025-1) | |
| Positive control | Adrenal gland+4 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 | Staining shows low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended HSD3B7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A10025-1) is accompanied by a published application in human glioma paraffin sections (PMC12963632).
| Sample | Paraffin-embedded human liver tissue; fixative not specified (datasheet A10025-1) |
| Fixation | Image fixative and duration unreported (datasheet A10025-1); 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 A10025-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10025-1) |
| Primary antibody | Rabbit anti-HSD3B7, 2-5 μg/ml (datasheet A10025-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10025-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10025-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSD3B7-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A10025-1); the glioma article does not specify retrieval conditions (PMC12963632).
Section 2
What Is the Expected HSD3B7 Staining Pattern?
In paraffin-section IHC, expect cytoplasmic HSD3B7 staining in glandular cells of the adrenal gland and several gastrointestinal tissues, kidney tubular cells, and lung macrophages (HPA: High in these cells). An intracellular pattern is consistent with its endoplasmic reticulum membrane annotation and two transmembrane segments (UniProt Q9H2F3 topology). HPA rates the tissue staining Approved but reports low agreement with RNA data and says external verification is pending (HPA: reliability).
| Cytoplasmic staining in kidney tubular cells, lung macrophages, or the listed glandular cells. | This matches the reported IHC distribution; HPA rates these cell groups High, except stomach glandular cells, which are Medium (HPA: tissue IHC). Score the named cells rather than assigning one intensity to an entire tissue. |
| A predominantly nuclear or extracellular deposit replaces the expected cytoplasmic pattern. | That compartment is inconsistent with the reported tissue pattern (HPA: cytoplasmic expression) and endoplasmic reticulum membrane annotation (UniProt Q9H2F3 topology). Check morphology, counterstain, and detection controls before interpreting it as HSD3B7. |
| Strong staining appears in cells reported as undetected, such as bronchial respiratory epithelium. | The result conflicts with the HPA tissue observation (HPA: bronchus respiratory epithelial cells Not detected). Cross-reactivity or detection-system activity is possible; the HPA rating and pending external verification make an independent check especially useful (HPA: reliability). |
| Color is spread across the section without clear cell boundaries or a cytoplasmic pattern. | Diffuse signal cannot establish cell-specific HSD3B7 expression. In chromogenic IHC, background from incomplete blocking, residual detection activity, or concentrated reagent can obscure the reported cell pattern (general IHC practice; HPA: cytoplasmic expression). |
| A listed High cell population has no detectable stain. | This disagrees with the HPA observation for that cell type (HPA: tissue IHC), but one negative slide does not prove absence. Review section quality, retrieval and detection conditions, and a working positive control (general IHC practice). |
💡Expected HSD3B7 appearanceCall a result positive when distinct cytoplasmic color marks the listed glandular cells, kidney tubules, or lung macrophages—typically High, with stomach glandular cells Medium (HPA: tissue IHC); isolated nuclear color, diffuse deposits, or strong signal in an HPA-undetected cell population warrant control checks (HPA: cytoplasmic expression and tissue IHC; general IHC practice).
| Tissue and cell selection | HPA reports High staining in adrenal, appendix, colon, duodenum, and small-intestine glandular cells, kidney tubules, and lung macrophages; stomach glandular cells are Medium (HPA: tissue IHC). Use the specified cell population when judging a control. |
| Strength of tissue evidence | The HPA tissue IHC entry is Approved, while its description reports low consistency between antibody staining and RNA expression and pending external verification (HPA: reliability). Treat a matching pattern as supporting evidence, especially when a new specimen differs. |
| RNA versus protein readout | Liver is tissue-enriched at the RNA level, but liver is not among the supplied positive IHC observations (HPA: RNA specificity; HPA: tissue IHC). RNA enrichment alone cannot specify the appearance or intensity of a liver IHC control. |
| Membrane topology and antibody access | HSD3B7 has two transmembrane segments at residues 289–309 and 311–331 (UniProt Q9H2F3 topology). That supports an intracellular membrane protein but does not identify the catalog antibody's epitope or establish an HSD3B7-specific retrieval setting. |
| Isoforms and processing | UniProt lists 2 isoforms and a chain spanning residues 1–369, with no signal peptide, propeptide, or annotated glycosylation sites (UniProt Q9H2F3). These annotations do not establish which isoforms the IHC antibody detects or predict different stain intensities. |
| IF/ICC Q: What localisation should be expected? | HPA reports an enhanced lipid-droplet localisation in ICC-IF, with images from Hep-G2, U2OS, MCF-7, and NIH 3T3 (HPA: subcellular; HPA: ICC Enhanced). This is a distinct assay observation; use the dedicated IF/ICC guide for its procedure. |
| Situation | Likely cause | Next action |
|---|---|---|
| No color in a chosen High-positive cell population. | The observed result conflicts with the HPA IHC report; a failed or weak staining run is one possible cause (HPA: tissue IHC; general IHC practice). | Verify that the target cells are present, then review the catalog antibody's IHC-P conditions, retrieval, dilution, and detection with a working positive section (general IHC practice). |
| Staining is mainly nuclear or extracellular. | The dominant compartment disagrees with HPA's cytoplasmic tissue pattern and UniProt's endoplasmic reticulum membrane location (HPA: tissue IHC; UniProt Q9H2F3). | Compare stain with cell morphology and counterstain; repeat with appropriate detection controls before scoring the unexpected compartment as positive (general IHC practice). |
| Bronchial respiratory epithelium or adipocytes stain strongly. | Those cells are reported Not detected by HPA; cross-reactivity or nonspecific color is possible, although an HPA-negative result is not proof of universal absence (HPA: tissue IHC; HPA: reliability). | Compare the suspect cells with a listed positive population on a controlled run and check a no-primary control; seek independent support for a persistent unexpected pattern (general IHC practice). |
| Color appears throughout tissue with little cellular definition. | Background can arise from staining or detection conditions and may hide a cytoplasmic pattern (general IHC practice; HPA: cytoplasmic expression). | Check reagent concentration, washes, blocking, and chromogen development against the validated IHC-P workflow; score only interpretable, cell-localized color (general IHC practice). |
| A no-primary control also develops color. | The signal can originate from endogenous detection activity or the detection reagents rather than primary-antibody binding (general chromogenic IHC practice). | Check endogenous-activity blocking and the detection reagents, then repeat the control alongside the specimen before interpreting HSD3B7 staining (general IHC practice). |
| IHC appears cytoplasmic while ICC-IF shows lipid droplets. | The reported readouts differ in compartment detail: tissue IHC is described as cytoplasmic, and ICC-IF as lipid-droplet localized (HPA: tissue IHC; HPA: subcellular). | Interpret each image within its assay and cell context; do not require paraffin-section chromogenic IHC to resolve the ICC-IF droplet pattern (HPA: tissue IHC; HPA: subcellular). |
Experiment evidence
IHC & IF images showing HSD3B7 staining
A10025-1 · IHCIHC analysis of HSD3B7 using anti-HSD3B7 antibody (A10025-1). HSD3B7 was detected in a paraffin-embedded section of human liver tissue. Heat
A10025-1 · IFIF analysis of HSD3B7 using anti-HSD3B7 antibody (A10025-1). HSD3B7 was detected in an immunocytochemical section of A549 cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000099377-HSD3B7/tissue/cerebral+cortex#imid_14551256
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000099377-HSD3B7/tissue/colon#imid_14551182
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000099377-HSD3B7/tissue/liver#imid_14551183
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000099377-HSD3B7/tissue/kidney#imid_14551200
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA015965; green = HSD3B7, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA015965; green = HSD3B7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSD3B7 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). Run cervix glandular cells as the negative tissue (HPA: Not detected in cervix glandular cells); on the colon slide, non-glandular stromal cells should remain at background staining and serve as an internal comparator, without assuming they are proven HSD3B7-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HSD3B7 in Hep-G2, U2OS, MCF-7, NIH 3T3, with annotated localisation: Lipid droplets (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU IHC caption: rabbit primary antibody). If available, use HSD3B7-knockout material as a biological negative; quench endogenous peroxidase in colon sections before HRP–DAB detection (selected-SKU IHC caption: HRP–DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state its fixative. The paraffin-section example uses heat-mediated EDTA retrieval at pH 8.0, but whether retrieval is required is unreported (selected-SKU IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; for colon IHC, check that glandular staining is distinct from endogenous peroxidase background (HPA: High in colon glandular cells; selected-SKU IHC caption: HRP–DAB detection).
HPA tissue IHC evidence for HSD3B7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HSD3B7 IHC Tips
Troubleshoot HSD3B7 staining by checking retrieval, cell identity, subcellular pattern, and controls before interpreting signal intensity.
What retrieval should I start with if HSD3B7 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A10025-1). The selected human liver image used that retrieval before incubation with 2 μg/ml antibody overnight at 4°C, so match those conditions when establishing a reference slide (datasheet A10025-1). If signal remains weak, vary heating and cooling within your validated IHC workflow while comparing adjacent sections; record tissue damage and background alongside signal (standard IHC practice). Keep a known staining section in each run, since a change in retrieval can alter apparent intensity without establishing that the new signal is HSD3B7 (standard IHC practice).
Could fixation explain inconsistent HSD3B7 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected human liver image identifies a paraffin-embedded section but does not state its fixative (datasheet A10025-1). Compare blocks with documented processing histories using the same EDTA pH 8.0 retrieval and staining run before assigning differences to biology (datasheet A10025-1; standard IHC practice). Include a reference section and inspect morphology, since uneven preservation or overprocessed tissue can complicate chromogenic scoring (standard IHC practice). Do not infer a preferred fixative or a specific fixation effect from the reported tissue staining patterns or from the protein’s membrane topology; neither supplies a fixation comparison (HPA tissue IHC; UniProt Q9H2F3 topology).
How should I interpret punctate versus diffuse cytoplasmic HSD3B7 staining?
HSD3B7 is annotated as an endoplasmic reticulum membrane protein with transmembrane segments at residues 289–309 and 311–331 (UniProt Q9H2F3 topology). Tissue IHC is described as cytoplasmic across several tissues, while subcellular IF reports enhanced lipid-droplet localisation; these observations support checking compartment patterns without expecting chromogenic IHC to resolve organelles reliably (HPA tissue IHC; HPA subcellular). Assess whether puncta occur within intact cells and recur on comparable sections, rather than treating isolated pigment or deposits as localisation (standard IHC practice). If compartment assignment matters, compare IHC with a separately controlled IF colocalisation experiment and report the methods independently (standard IHC/IF practice).
Can this antibody distinguish HSD3B7 isoforms or membrane-facing epitopes?
The record lists 2 HSD3B7 isoforms, but supplies no isoform-specific epitope map for the catalog antibody (UniProt Q9H2F3 isoforms; datasheet A10025-1). Its 2 annotated transmembrane segments lie at residues 289–309 and 311–331; that topology alone does not locate the antibody epitope or establish which membrane face it recognizes (UniProt Q9H2F3 topology). Check the antibody’s documented immunogen or epitope before claiming isoform-selective staining, and use an independently validated reagent if that distinction is essential (standard IHC practice). Report staining as HSD3B7-associated until specificity controls and epitope information support a narrower interpretation (standard IHC practice).
How can IF help resolve ambiguous HSD3B7 IHC localisation?
Use IF as a separate localisation check, pairing HSD3B7 with a marker for the cell type under study, such as hepatocytes in liver or macrophages in lung (HPA liver RNA specificity; HPA: high in lung macrophages; standard IF practice). Select a fluorophore and detection channel that separate specific signal from tissue autofluorescence, and inspect single-channel controls before calling colocalisation (standard IF practice). Because HSD3B7 has membrane-spanning regions but the antibody epitope’s orientation is unspecified, choose permeabilisation only after establishing which side of the membrane the epitope occupies (UniProt Q9H2F3 topology; standard IF practice). The reported lipid-droplet IF pattern can guide comparison, but does not validate an IF protocol for this catalog antibody (HPA subcellular; datasheet A10025-1).
What should I check when DAB obscures HSD3B7-positive cells?
The selected liver IHC used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB detection (datasheet A10025-1). When background rises, compare a primary-omission control and a matched section with adjusted primary concentration or incubation, changing one variable at a time (standard IHC practice). Apply an appropriate endogenous-peroxidase block and inspect the omission control before attributing brown signal to HSD3B7; these are general chromogenic IHC safeguards, not HSD3B7-specific findings (standard IHC practice). Evaluate tissue edges, folds, deposits, and counterstain separately from signal within intact cells, using matched development conditions across sections (standard IHC practice).
How should I score HSD3B7 IHC across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive cells and intensity or calculate an H-score from those values (standard IHC practice). Normalize counts to the number of eligible intact cells, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Keep retrieval, antibody concentration, DAB development, imaging, and positivity thresholds consistent across compared sections; the selected image used EDTA at pH 8.0 and 2 μg/ml primary antibody (datasheet A10025-1; standard IHC practice). Score cell types separately because reported staining includes glandular cells, kidney tubules, and lung macrophages (HPA tissue IHC).
Which findings make an HSD3B7-positive IHC result convincing?
A convincing result places signal within intact cells, follows a reproducible cytoplasmic pattern, and agrees with a controlled reference section; HSD3B7 is annotated at the endoplasmic reticulum membrane (standard IHC practice; UniProt Q9H2F3 localisation). Check cell identity: HPA reports high staining in lung macrophages and kidney tubule cells, but no detection in adipocytes or lymph-node germinal-center cells (HPA tissue IHC). Treat isolated nuclear staining, edge staining, necrotic areas, and signal persisting after primary omission as possible artefact or endogenous-enzyme background pending investigation (standard IHC practice). Interpret agreement cautiously because the tissue IHC entry notes low consistency with RNA expression and pending external verification (HPA tissue IHC).
Boster reagents
Best HSD3B7 / 3 beta-hydroxysteroid dehydrogenase type 7 IHC Antibodies
A10025-1 has IHC data from human liver paraffin sections (catalog IHC image caption) and IF data from A549 cells (catalog IF image caption); listed reactivity is human (catalog: reactivity).
Real IHC data
Anti-HSD3B7 Antibody ®
Cat # A10025-1
A10025-1 is listed for human IHC and IF/ICC (catalog: applications and reactivity), with an IHC image from a paraffin-embedded human liver section (catalog IHC image caption). Its IF image shows staining in A549 cells (catalog IF image caption).
Which to pick: Choose A10025-1 for human paraffin-section IHC: its own image caption documents liver staining after heat retrieval in EDTA at pH 8.0; the fixative is unreported (catalog IHC image caption). For IF/ICC, A10025-1 is listed for both applications and shown staining A549 cells at 5 μg/ml; clonality is unreported (catalog: applications and clone; catalog IF image caption). No cross-species choice is supported because A10025-1 lists human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H2F3 (3BHS7_HUMAN, 3 beta-hydroxysteroid dehydrogenase type 7).
- Human Protein Atlas. HSD3B7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. HSD3B7 subcellular location (ICC-IF): Localized to the lipid droplets..
- Human Protein Atlas. HSD3B7 antibody validation summary (3 antibodies).
- Lipid metabolism classification of gliomas. Scientific reports 2026 — PMC12963632.
- PubMed PMID:11067870 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSD3B7 (3 beta-hydroxysteroid dehydrogenase type 7). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane localization (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10025-1). Validated IHC antibody: A10025-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver tissue; fixative not specified (datasheet A10025-1); 2. Fixation: Image fixative and duration unreported (datasheet A10025-1); 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 A10025-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10025-1); 8. Primary antibody: Rabbit anti-HSD3B7, 2-5 μg/ml (datasheet A10025-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10025-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining shows low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt Q9H2F3, HPA tissue IHC · Approved, HPA ICC-IF · lipid droplets., Datasheet A10025-1, PMC12963632. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSD3B7/immunohistochemistry-ihc-guide.