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HSDL2 / Hydroxysteroid dehydrogenase-like protein 2 · IHC design guide
Design Immunohistochemistry for HSDL2
Plan HSDL2 IHC in paraffin sections around granular cytoplasmic staining (HPA tissue IHC). Compare high-staining adrenal glandular cells with low-staining skeletal myocytes (HPA tissue IHC), and optimize the catalog antibody within 2–5 μg/ml (datasheet A11317-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSDL2 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 cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11317-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Widespread staining limits tissue-negative controls (HPA tissue IHC) | |
| Regulation | Expression regulation not established (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended HSDL2 IHC & IF Protocols
The catalog antibody protocol is accompanied by 3 published HSDL2 IHC methods for mouse hippocampus, human ovarian tissue, and human thyroid tissue (datasheet A11317-1; PMC13369707; PMC6029517; PMC6549096).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A11317-1) |
| Fixation | Image fixative and duration unreported (datasheet A11317-1); 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 A11317-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11317-1) |
| Primary antibody | Rabbit anti-HSDL2, 2-5 μg/ml (datasheet A11317-1) |
| Primary incubation | Overnight at 4 °C (datasheet A11317-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11317-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSDL2-positive staining in glandular cells of adrenal gland (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 EDTA pH 8.0 retrieval for the catalog antibody (datasheet A11317-1); citrate pH 6.1 was used in the thyroid study (PMC6549096).
Section 2
What Is the Expected HSDL2 Staining Pattern?
HSDL2 should show granular cytoplasmic staining in tissue sections (HPA: granular cytoplasmic expression in all tissues). High staining is reported in adrenal and colon glandular cells, bone marrow hematopoietic cells, and esophageal squamous epithelial cells (HPA: High). The tissue IHC profile has Enhanced reliability (HPA: Enhanced). UniProt places HSDL2 in peroxisomes and mitochondria and reports no transmembrane segment (UniProt Q6YN16: subcellular location; topology).
| Granular cytoplasmic stain in colon glandular cells or bone marrow hematopoietic cells. | This fits the reported tissue pattern (HPA: granular cytoplasmic expression; High in both cell populations). Judge granularity within the expected cells alongside the counterstain; a uniformly dark section is harder to interpret (general IHC practice). |
| Predominantly nuclear or sharply cell-surface staining, with little granular cytoplasmic signal. | This differs from the reported IHC pattern (HPA: granular cytoplasmic expression) and organelle locations (UniProt Q6YN16: peroxisome; mitochondrion). Treat a compartment-only result as suspect and check a positive control before assigning HSDL2 localisation (general IHC practice). |
| Strong stain mainly outside the reported high-staining cell population, such as colon stroma rather than glandular cells. | The distribution does not match the reported high-staining population (HPA: High in colon glandular cells). Cross-reactivity or endogenous chromogen-generating activity may contribute (general IHC practice); assess controls before calling it specific. HSDL2 is described as ubiquitous (UniProt Q6YN16: tissue specificity), so staining elsewhere is not automatically false. |
| Diffuse haze covers cells and surrounding section, obscuring cytoplasmic granules. | The appearance cannot establish the reported granular pattern (HPA: tissue IHC profile). Excess background can arise from detection or blocking conditions (general IHC practice). Compare a primary-antibody-omission control, then adjust the general IHC workflow before scoring cells. |
| No stain in an otherwise interpretable adrenal gland or duodenum section. | This conflicts with reported high staining in glandular cells at those sites (HPA: High). First check that the expected cells are present and the detection control worked (general IHC practice). A blank section alone cannot establish that HSDL2 is absent. |
💡Expected HSDL2 appearanceCall a section positive when the expected cells show discernible granular cytoplasmic staining, including strong signal in reported high-staining populations (HPA: tissue IHC profile); a nuclear-only, surface-only, or uniform diffuse stain is discordant with that profile (HPA: granular cytoplasmic expression).
| Tissue and cell selection | Choose a reported high-staining population to judge assay performance, such as adrenal glandular cells or bone marrow hematopoietic cells (HPA: High). Skeletal-muscle myocytes and lymph-node germinal-center cells are reported Low (HPA: Low), so weak staining there is a poor stand-alone failure test. |
| Evidence strength and limits | Tissue staining has Enhanced reliability, and both listed antibodies have Enhanced IHC status (HPA: tissue reliability; HPA042890 and HPA050453 IHC). This supports the reported pattern but does not prove that every stain in an individual section is specific (general IHC interpretation). |
| Organelle location and topology | UniProt lists peroxisomes and mitochondria and no transmembrane segment (UniProt Q6YN16: subcellular location; topology). Expect a cytoplasmic organelle-associated appearance in IHC (HPA: granular cytoplasmic expression); chromogenic tissue staining alone does not resolve which of the two organelles carries each granule. |
| Isoforms and processing | Two isoforms are listed, with no signal peptide or propeptide and a 1–418 chain reported (UniProt Q6YN16: isoforms; processing). The payload does not map antibody epitopes to either isoform, so an unusual stain cannot be assigned to a specific isoform or cleavage event from these data. |
| IF/ICC Q&A: what localisation should be expected? | HPA reports enhanced mitochondrial localisation by ICC-IF (HPA: subcellular summary; main location). UniProt also lists peroxisomes (UniProt Q6YN16: subcellular location). Use this as a localisation cross-check; the IF/ICC workflow belongs in its own guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in high-staining glandular cells. | The result disagrees with the tissue reference (HPA: High in adrenal gland and duodenum glandular cells); the specific failed step is not established. | Confirm the expected cells are present, inspect a positive control and detection reagents, then review retrieval and antibody dilution using the assay instructions (general IHC practice). |
| Low signal in skeletal-muscle myocytes or lymph-node germinal-center cells. | Low staining is reported for these populations (HPA: Low); their faint appearance alone does not diagnose assay failure. | Compare with a reported high-staining population on a separately controlled section (HPA: High in colon glandular cells; general IHC practice). |
| Nuclear-only or membrane-rim signal dominates. | The compartment differs from granular cytoplasmic tissue staining (HPA: tissue IHC profile) and from the annotated organelle locations (UniProt Q6YN16: subcellular location). | Recheck morphology and compare with a positive control; review blocking and detection controls before interpreting the discordant stain as HSDL2 (general IHC practice). |
| Strong colour appears across the section, including areas without the expected cell pattern. | Diffuse colour obscures the reported granular cytoplasmic appearance (HPA: tissue IHC profile); endogenous activity or detection background may be involved (general IHC practice). | Run a primary-antibody-omission control and review endogenous-activity blocking, washing, and chromogen development under the established IHC workflow (general IHC practice). |
| Unexpected cell population stains more strongly than the reported high-staining cells. | This may reflect cross-reactivity or background (general IHC practice), but HSDL2 is described as ubiquitous (UniProt Q6YN16: tissue specificity). The population alone cannot settle specificity. | Check the cell identity and compartment, compare the reported high-staining population, and use control staining before scoring the unexpected signal (HPA: tissue IHC profile; general IHC practice). |
| IHC granules cannot be confidently assigned to mitochondria. | Tissue IHC reports a granular cytoplasmic pattern (HPA: tissue IHC profile), while ICC-IF reports enhanced mitochondrial localisation and UniProt also lists peroxisomes (HPA: subcellular summary; UniProt Q6YN16: subcellular location). | Report the IHC result as granular cytoplasmic staining; consult the separate IF/ICC localisation guide when organelle assignment is required (HPA: tissue IHC and ICC-IF profiles). |
Experiment evidence
IHC & IF images showing HSDL2 staining
A11317-1 · IHCIHC analysis of HSDL2 using anti-HSDL2 antibody (A11317-1). HSDL2 was detected in a paraffin-embedded section of human colon adenocarcinoma
A11317-1 · IHCIHC analysis of HSDL2 using anti-HSDL2 antibody (A11317-1). HSDL2 was detected in a paraffin-embedded section of human liver cancer tissue.
A11317-1 · IFIF analysis of HSDL2 using anti-HSDL2 antibody (A11317-1). HSDL2 was detected in an immunocytochemical section of A549 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000119471-HSDL2/tissue/cerebral+cortex#imid_11833603
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000119471-HSDL2/tissue/colon#imid_11833545
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000119471-HSDL2/tissue/liver#imid_11833538
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000119471-HSDL2/tissue/kidney#imid_11833531
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA042890; green = HSDL2, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA042890; green = HSDL2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSDL2 IHC & IF
🧪Start with colon and score glandular cells, which HPA rates High for HSDL2 (HPA: colon glandular cells High). HPA detects HSDL2 in all 45 scored tissues, so there is no supported negative tissue; use the no-primary and isotype controls to define background, and treat adjacent nonglandular cells as an internal comparison rather than confirmed negative cells (HPA: no negative tissue rows; UniProt: ubiquitous).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: HSDL2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HSDL2 in Rh30, U2OS, A-431, U-251MG, with annotated localisation: Mitochondria (enhanced) (HPA subcellular).
Technical controls: Run a secondary-only slide with no primary antibody and a matched rabbit IgG isotype control; a validated HSDL2 knockout specimen or peptide-blocked primary can provide a biological specificity control (selected-SKU caption: rabbit primary antibody; standard IHC practice). Check endogenous peroxidase signal before interpreting DAB staining in colon sections (selected-SKU caption: colon section with HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected-SKU caption: EDTA retrieval at pH 8.0). There is no supplied comparison showing frozen sections or IF to be easier; HPA does provide ICC-IF images with enhanced mitochondrial localization, while endogenous peroxidase can complicate colon DAB scoring (HPA: mitochondria enhanced; selected-SKU caption: colon HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for HSDL2
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: HSDL2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced HSDL2 IHC Tips
Troubleshoot HSDL2 staining in paraffin sections by checking retrieval, cellular pattern, controls, and scoring against the available tissue evidence.
How should I adjust retrieval when HSDL2 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A11317-1). The selected tissue image used that condition, but its caption gives no heating duration or fixative (caption A11317-1). Keep the buffer constant while comparing two controlled heating durations, such as 10 and 20 minutes, with matched sections and cooling conditions (standard IHC practice). If staining remains weak, test citrate at pH 6.0 as a fallback and compare both signal and tissue damage (standard IHC practice). Judge improvement by granular cytoplasmic staining in intact cells, rather than stronger edge staining alone (HPA tissue IHC; standard IHC practice).
Could fixation explain inconsistent HSDL2 staining between paraffin blocks?
Target-specific fixation sensitivity for HSDL2 is unknown from the supplied evidence (caption A11317-1). The selected image identifies a paraffin-embedded colon adenocarcinoma section but does not report its fixative or fixation time (caption A11317-1). Record the fixative, tissue thickness, time before fixation, and fixation duration for each block before comparing staining (standard IHC practice). On otherwise matched samples, compare documented fixation intervals such as 6 and 24 hours, then apply the same EDTA pH 8.0 retrieval (standard IHC practice; datasheet A11317-1). Treat any difference as a result of that controlled comparison, not as an established HSDL2 fixation effect.
Does diffuse cytoplasmic HSDL2 staining fit the expected pattern?
Expect a granular cytoplasmic pattern in tissue IHC (HPA tissue IHC). HSDL2 is annotated in peroxisomes and mitochondria, while the HPA subcellular assessment identifies mitochondria as its enhanced main location (UniProt Q6YN16 subcellular; HPA subcellular). Chromogenic sections generally cannot distinguish those organelles reliably, so score cellular distribution and granularity rather than assigning individual DAB deposits to one organelle (standard IHC practice). Compare glandular cells in colon, which show high staining in the tissue reference, with adjacent intact cells on the same section (HPA: High in colon glandular cells). Uniform nuclear colour, necrotic deposits, or staining confined to cut edges warrants review of controls and morphology (standard IHC practice).
How could epitope choice affect HSDL2 tissue staining?
HSDL2 has 2 annotated isoforms, and its SCP2 domain spans residues 306–415 (UniProt Q6YN16 isoforms and domains). The supplied antibody caption does not specify its immunogen or epitope, so it cannot establish recognition of both isoforms (caption A11317-1). Obtain the antibody’s epitope information before attributing staining differences between specimens to isoform expression (standard IHC practice). Reported modified residues include positions 42, 116, and 318, but the payload does not show that these modifications change tissue staining (UniProt Q6YN16 modified residues). If an epitope maps near one of those sites, compare retrieval conditions and an independent epitope before proposing masking as the explanation (standard IHC practice).
How should I assess HSDL2 by multiplex immunofluorescence?
For a colon section, pair HSDL2 with a glandular epithelial marker to identify the expected high-staining cell population (HPA: High in colon glandular cells; standard IF practice). Put the weaker target signal in a far-red channel when tissue autofluorescence is prominent, and check unstained tissue in every channel (standard IF practice). HSDL2 lacks an annotated transmembrane segment, but the supplied record does not locate the antibody epitope relative to organelle membranes (UniProt Q6YN16 topology; caption A11317-1). Compare mild permeabilisation, such as 0.1% detergent for 5 minutes, with an unpermeabilised section and inspect morphology (standard IF practice). Confirm puncta with a mitochondrial marker while allowing for peroxisomal localisation; this IF workflow has no selected-antibody validation in the supplied caption (HPA subcellular; UniProt Q6YN16 subcellular; caption A11317-1).
What should I check when DAB obscures HSDL2 staining?
The selected paraffin-section image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase-based DAB detection (caption A11317-1). Begin with a no-primary section to assess secondary-reagent and endogenous-enzyme signal, and include a peroxidase block in the chromogenic workflow (standard IHC practice). If diffuse brown colour persists, shorten DAB development and compare a primary-antibody dilution series while keeping section thickness and retrieval fixed (standard IHC practice). Inspect pigment and necrotic areas separately from intact cells because deposits there can mimic specific staining (standard IHC practice). Retain conditions that preserve granular cytoplasmic signal without a similar no-primary pattern (HPA tissue IHC; standard IHC practice).
How can I score HSDL2 across heterogeneous paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define viable, interpretable cells and exclude folds, necrosis, and cut edges before scoring (standard IHC practice). For each region, report the percentage of positive cells and an H-score calculated as the sum of percentages at intensities 0–3 multiplied by intensity, yielding 0–300 (standard IHC practice). Record granular cytoplasmic signal as the scoring feature, using the same positivity threshold and DAB exposure across specimens (HPA tissue IHC; standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or stained area to viable tissue area in mm², rather than whole-slide area (standard IHC practice). Separate glandular and other compartments where relevant because colon glandular cells have high reference staining (HPA: High in colon glandular cells).
When is a brown HSDL2 signal likely to be artefactual?
A plausible positive is granular cytoplasmic staining in intact cells, consistent with the tissue reference and HSDL2 organelle annotations (HPA tissue IHC; UniProt Q6YN16 subcellular). High staining in colon glandular cells supports that pattern, whereas an isolated nuclear-only signal calls for further validation (HPA: High in colon glandular cells; HPA tissue IHC). Check whether colour concentrates at section edges, in necrosis, or over pigment before calling a weak focus positive (standard IHC practice). Compare a no-primary section and a peroxidase-blocked section to distinguish reagent or endogenous-enzyme colour from antibody-dependent DAB signal (standard IHC practice). Interpret differences between specimens only after retrieval, fixation records, and scoring thresholds are matched (standard IHC practice).
Boster reagents
Best HSDL2 / Hydroxysteroid dehydrogenase-like protein 2 IHC Antibodies
The catalog antibody has IHC images from human paraffin sections and IF images from A549 cells and a human paraffin section (catalog image captions); listed reactivity is human, mouse and rat (catalog).
Real IHC data
Anti-HSDL2 Antibody ®
Cat # A11317-1
A11317-1 has IHC images from paraffin sections of human colon adenocarcinoma, liver cancer, lung cancer and pancreas ductal adenocarcinoma (catalog IHC image captions). Its IF images show A549 cells and a paraffin section of human intestinal cancer (catalog IF image captions).
Which to pick: Choose A11317-1 for tissue IHC: its paraffin-section captions report EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (catalog IHC image captions). The same SKU is listed for IF and ICC, with 5 μg/ml listed for each; its IF images show A549 cells and human intestinal cancer tissue (catalog applications, dilution listing and IF image captions). It lists human, mouse and rat reactivity, but the supplied IHC images show human tissue only, and clonality is unreported (catalog reactivity, IHC image captions and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6YN16 (HSDL2_HUMAN, Hydroxysteroid dehydrogenase-like protein 2).
- Human Protein Atlas. HSDL2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HSDL2 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. HSDL2 antibody validation summary (2 antibodies).
- SIRT5-mediated HSDL2 desuccinylation promotes diabetic-associated cognitive dysfunction via disrupting mitochondrial TRAP1. Cell biology and toxicology 2026 — PMC13369707.
- Role of Hydroxysteroid Dehydrogenase-Like 2 (HSDL2) in Human Ovarian Cancer. Medical science monitor : international medical journal of experimental and clinical research 2018 — PMC6029517.
- Down-regulated HSDL2 expression suppresses cell proliferation and promotes apoptosis in papillary thyroid carcinoma. Bioscience reports 2019 — PMC6549096.
- HSDL2 Suppresses Epileptic Seizures Through Phosphorylation-Dependent Modulation of the PSD95-NMDAR Signaling Axis. CNS neuroscience & therapeutics 2026 — PMC12971609.
- PubMed PMID:12834046 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSDL2 (Hydroxysteroid dehydrogenase-like protein 2). Expected localisation Granular cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11317-1). Validated IHC antibody: A11317-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A11317-1); 2. Fixation: Image fixative and duration unreported (datasheet A11317-1); 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 A11317-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11317-1); 8. Primary antibody: Rabbit anti-HSDL2, 2-5 μg/ml (datasheet A11317-1); 9. Primary incubation: Overnight at 4 °C (datasheet A11317-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Widespread staining limits tissue-negative controls (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: Expression regulation not established (UniProt). Evidence: UniProt Q6YN16, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A11317-1, PMC13369707, PMC6029517. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSDL2/immunohistochemistry-ihc-guide.