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HSD11B2 / 11-beta-hydroxysteroid dehydrogenase type 2 · IHC design guide
Design Immunohistochemistry for HSD11B2
Plan paraffin-section HSD11B2 IHC around cytoplasmic staining in kidney collecting ducts and gastrointestinal cell subsets (HPA tissue IHC). Start the catalog antibody RP1094 at 0.5–1 μg/ml and compare cell-level staining with controls (datasheet: RP1094; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSD11B2 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 staining in select tissue cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in renal tubules and GI glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet RP1094) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image RP1094) | |
| Caveat | Restricted positive cell types can be missed in bulk scoring (HPA tissue IHC) | |
| Regulation | Tissue-dependent expression (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one 1–405 chain (UniProt) |
Section 1
Recommended HSD11B2 IHC & IF Protocols
The catalog antibody protocol is paired with published HSD11B2 IHC methods for dog placenta and colorectal adenoma and normal mucosa (PMC12561314; PMC9832797).
| Sample | Paraffin-embedded Mouse Pancreas tissue; fixative not specified (datasheet RP1094) |
| Fixation | Image fixative and duration unreported (datasheet RP1094); 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 RP1094); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet RP1094) |
| Primary antibody | Rabbit anti-HSD11B2, 0.5-1μg/ml (datasheet RP1094) |
| Primary incubation | Overnight at 4 °C (datasheet RP1094) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet RP1094) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSD11B2-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in few tissues, including cells in renal tubules and glandular cells in gastrointestinal tract and salivary gland. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet RP1094); the published methods used EDTA pH 9.0 or citrate pH 6.0 (PMC12561314; PMC9832797).
Section 2
What Is the Expected HSD11B2 Staining Pattern?
HSD11B2 is associated with microsomes and the endoplasmic reticulum, with no annotated transmembrane segment (UniProt P80365 topology). In paraffin-section IHC, expect cytoplasmic staining in selected cells, especially kidney collecting ducts, intestinal endocrine cells, duodenal goblet cells, and placental syncytiotrophoblast cell bodies (HPA tissue IHC: High). The tissue pattern has Enhanced reliability, reflecting agreement between staining and RNA data (HPA tissue IHC: Enhanced).
| Distinct cytoplasmic staining in kidney collecting ducts, with nearby cells less conspicuous. | This fits a strong positive result: collecting ducts are rated High, while HPA describes cytoplasmic expression in only a few tissues (HPA tissue IHC: High; HPA tissue profile). Judge the named cells separately from the overall kidney section; uniform staining throughout the section would need scrutiny. |
| Predominantly nuclear or sharply cell-surface staining, without a convincing cytoplasmic component. | Treat this as a compartment mismatch. UniProt places HSD11B2 in microsomes and endoplasmic reticulum, and HPA reports cytoplasmic tissue staining (UniProt P80365 subcellular location; HPA tissue profile). Check morphology and controls before interpreting the signal as target-specific. |
| Strong signal in a cell population listed as not detected, such as bronchial respiratory epithelium. | This conflicts with that cell-level HPA observation (HPA tissue IHC: Bronchus, Not detected). Consider antibody cross-reactivity or endogenous chromogenic detection activity; compare the stained section with an appropriate negative detection control (standard IHC practice). |
| Weak color spread across cells, extracellular spaces, or much of the section. | A diffuse pattern does not match HPA's selective cytoplasmic profile (HPA tissue profile). Assess background using a control that omits the primary antibody, then review blocking, antibody concentration, washing, and chromogen development (standard IHC practice). |
| No convincing signal in kidney collecting ducts or another documented High population. | A blank known-positive population makes this run difficult to interpret, because those cells show High staining in the HPA record (HPA tissue IHC: Kidney; Placenta). Check that the expected cells are present, then review the antibody's validated IHC-P conditions and the detection controls (standard IHC practice). |
💡Expected HSD11B2 appearanceCall the result positive when staining is distinct and cytoplasmic in documented High populations such as kidney collecting ducts; broad nuclear staining or strong staining in HPA Not detected cells warrants investigation (HPA tissue IHC: Kidney, High; Bronchus, Not detected; HPA tissue profile; UniProt P80365 subcellular location).
| Which cells provide the clearest tissue check? | Kidney collecting ducts, placental syncytiotrophoblast cell bodies, duodenal goblet cells, and endocrine cells in appendix, colon, rectum, and small intestine are rated High (HPA tissue IHC). Select a control whose named cell population is visible; a tissue name alone does not identify every cell as positive. |
| How broadly should staining be expected? | HPA describes cytoplasmic expression in few tissues and rates endometrial stromal cells Medium, while several listed cell populations are Not detected (HPA tissue IHC). UniProt reports expression across additional tissues, but that statement does not assign an IHC intensity to each cell type (UniProt P80365 tissue specificity). |
| How much confidence does antibody validation provide? | HPA rates tissue IHC reliability Enhanced and lists HPA042186, HPA056385, and CAB032443 as IHC Enhanced (HPA tissue IHC; HPA antibodies). This supports the reported pattern, but a new section still needs its own staining and detection controls (standard IHC practice). |
| Does ICC-IF imply the same visual pattern as tissue IHC? | In ICC-IF, HPA reports supported vesicle localization and images from CACO-2 and RT-4 (HPA subcellular ICC-IF). For paraffin-section IHC, use the tissue record's cytoplasmic description; the ICC-IF observation does not specify an IHC staining intensity or an IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive kidney section appears negative. | The expected collecting-duct cells may be absent from the viewed area, or the staining run may have failed (HPA tissue IHC: Kidney, High; standard IHC practice). | Confirm collecting ducts by morphology; check the IHC-validated antibody's stated conditions and the detection control before calling the sample negative (standard IHC practice). |
| Nuclei carry the strongest signal. | This disagrees with the cytoplasmic tissue profile and microsomal/endoplasmic-reticulum assignment (HPA tissue profile; UniProt P80365 subcellular location). | Compare with a positive tissue and a primary-omission control; interpret persistent nuclear-only staining cautiously (standard IHC practice). |
| A HPA Not detected cell population stains strongly. | Cross-reactivity or endogenous chromogenic detection activity is possible; HPA's Not detected designation applies to the specified cells (HPA tissue IHC; standard IHC practice). | Check a primary-omission control and apply the detection system's endogenous-activity control as appropriate (standard IHC practice). |
| Color is diffuse across the section. | Nonspecific background can arise during blocking, antibody incubation, washing, or chromogen development (standard IHC practice). | Compare with a primary-omission control and review those steps using the antibody's IHC-P instructions (standard IHC practice). |
| A positive tissue shows only faint staining. | The selected cells may have a lower reported level, or the run may be underdeveloped (HPA tissue IHC: Endometrial stroma, Medium; standard IHC practice). | Verify the cell type and compare it with a documented High population on a successfully stained control section (HPA tissue IHC; standard IHC practice). |
| ICC-IF looks punctate while IHC looks broadly cytoplasmic. | HPA describes supported vesicles in ICC-IF and cytoplasmic staining in tissue IHC; these are observations from different preparations (HPA subcellular ICC-IF; HPA tissue profile). | Evaluate each result against its own HPA localization record and controls; do not require identical visual texture across the two applications (standard IHC/IF practice). |
Experiment evidence
IHC & IF images showing HSD11B2 staining
RP1094 · IHCIHC analysis of HSD11B2 using anti-HSD11B2 antibody (RP1094). HSD11B2 was detected in a paraffin-embedded section of Mouse Pancreas tissue.
RP1094 · IHCIHC analysis of HSD11B2 using anti-HSD11B2 antibody (RP1094). HSD11B2 was detected in a paraffin-embedded section of Mouse Pancreas tissue.
RP1094 · IFIF analysis of HSD11B2 using anti-HSD11B2 antibody (RP1094). HSD11B2 was detected in a paraffin-embedded section of human placenta tissue. H
A01613 · IHCIHC analysis of HSD11B2 using anti-HSD11B2 antibody (A01613). HSD11B2 was detected in a paraffin-embedded section of human appendix cancer t
A01613 · IHCIHC analysis of HSD11B2 using anti-HSD11B2 antibody (A01613). HSD11B2 was detected in a paraffin-embedded section of human breast cancer tis
A01613 · IFIF analysis of HSD11B2 using anti-HSD11B2 antibody (A01613). HSD11B2 was detected in a paraffin-embedded section of human placenta tissue. H
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176387-HSD11B2/tissue/cerebral+cortex#imid_16568883
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176387-HSD11B2/tissue/colon#imid_16568825
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176387-HSD11B2/tissue/liver#imid_16568810
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000176387-HSD11B2/tissue/kidney#imid_16568251
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA056385; green = HSD11B2, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA056385; green = HSD11B2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSD11B2 IHC & IF
🧪Run kidney first and expect staining in collecting ducts (HPA: High in kidney collecting ducts). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); compare neighboring non-collecting-duct cells on the kidney slide with the collecting ducts, without assuming every neighboring cell is negative (HPA: High in kidney collecting ducts).
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 HSD11B2 in CACO-2, RT-4, with annotated localisation: Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched nonimmune rabbit IgG isotype control (selected RP1094 tissue-IHC caption: rabbit primary); use HSD11B2 knockout tissue or a validated peptide block as a biological specificity control (standard IHC practice). Block endogenous peroxidase in the kidney section before HRP/DAB detection (selected RP1094 tissue-IHC caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: The selected RP1094 paraffin-section caption reports heat retrieval in EDTA at pH 8.0 for mouse pancreas, but its fixative is unreported; no target-specific fixation window or fixation effect is reported in the supplied evidence (selected RP1094 tissue-IHC caption). That caption supports testing retrieval for paraffin IHC, but does not establish that frozen sections or IF are easier; HPA reports vesicular ICC-IF localization in CACO-2 and RT-4 (HPA subcellular). For kidney IHC with HRP/DAB, endogenous peroxidase can create background unless blocked (standard IHC practice).
HPA tissue IHC evidence for HSD11B2
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 | Goblet cells | High | Protein (IHC) | HPA → |
| Kidney | Collecting ducts | High | Protein (IHC) | HPA → |
| Placenta | Syncytiotrophoblasts - cell body | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HSD11B2 IHC Tips
Troubleshoot HSD11B2 staining in paraffin sections by checking retrieval, cell identity, compartment, controls, and scoring before interpreting chromogenic signal.
What should I change if HSD11B2 staining is weak after antigen retrieval?
Begin with heat-mediated EDTA pH 8.0 retrieval for paraffin sections (datasheet RP1094). Use the catalog antibody at 1 μg/ml overnight at 4°C, and compare a kidney collecting-duct control with the test section in the same staining run (datasheet RP1094; HPA tissue IHC). If signal remains weak, vary heating duration on matched sections before trying another retrieval buffer, while keeping section thickness and development time comparable (standard IHC practice). Judge improvement by cytoplasmic staining in expected cells: HSD11B2 is assigned to microsomes and endoplasmic reticulum, and tissue staining is reported as cytoplasmic (UniProt P80365; HPA tissue IHC).
How can I troubleshoot fixation-related loss of HSD11B2 staining?
The selected RP1094 image describes a paraffin-embedded mouse pancreas section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet RP1094). Record fixative, fixation duration, tissue thickness, and processing history for each block before comparing staining intensity across specimens (standard IHC practice). When a block stains weakly, run it beside a consistently processed positive control and keep EDTA pH 8.0 retrieval and the 1 μg/ml primary concentration constant (datasheet RP1094; standard IHC practice). Compare well-preserved interior cells with damaged or poorly preserved regions, and avoid attributing a difference to fixation until matched processing controls support that conclusion (standard IHC practice).
Where should convincing HSD11B2 staining appear in tissue sections?
Look for cytoplasmic staining within the appropriate cells, since tissue IHC is described as cytoplasmic and HSD11B2 is assigned to microsomes and endoplasmic reticulum (HPA tissue IHC; UniProt P80365). Kidney collecting ducts and placental syncytiotrophoblast cell bodies are reported as high-staining sites and can anchor a same-run comparison (HPA tissue IHC). At 40× objective magnification, compare the distribution within cells against the hematoxylin counterstain, recording any predominantly nuclear or extracellular deposit separately (standard IHC practice). A vesicular-looking pattern alone is insufficient to assign tissue cell identity, because vesicles are reported from ICC/IF imaging while tissue IHC requires morphological context (HPA subcellular; standard IHC practice).
Could an isoform or inaccessible epitope explain discordant HSD11B2 staining?
The supplied protein record lists a single 405-aa chain and 0 annotated isoforms, so an annotated isoform switch does not explain discordant staining (UniProt P80365). It also lists no transmembrane segment, glycosylation sites, or modified residues, but gives no antibody epitope; these annotations cannot establish epitope accessibility in a section (UniProt P80365; datasheet RP1094). Compare adjacent sections under the stated EDTA pH 8.0 retrieval and a controlled change in retrieval duration before assigning epitope loss (datasheet RP1094; standard IHC practice). Document the antibody's immunogen or mapped epitope if available, and interpret a negative result against a same-run positive tissue control (standard IHC practice).
How should I assess HSD11B2 in a multiplex IF experiment?
For the separate IF/ICC workflow, pair HSD11B2 with a marker that identifies the expected cell population, such as collecting-duct cells when examining kidney, and confirm cell boundaries in a nuclear channel (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores after inspecting unstained tissue autofluorescence, and favor a channel with low background for the weaker target (standard IF practice). Match permeabilisation to the antibody's mapped epitope and intracellular access, since HSD11B2 is assigned to microsomes and endoplasmic reticulum but the supplied record does not locate this antibody's epitope (UniProt P80365; standard IF practice). Assess each channel alone before interpreting overlap; vesicular localisation has been reported in ICC/IF images (HPA subcellular; standard IF practice).
How do I separate HSD11B2 signal from chromogenic background?
Compare the stained section with a no-primary control and a same-run positive control, then inspect whether DAB follows cell morphology rather than pooling at folds or edges (standard IHC practice). The RP1094 image used 10% goat serum blocking, a peroxidase-linked secondary, and DAB development, providing a documented starting point for that antibody (datasheet RP1094). Include a peroxidase block before chromogen development and shorten DAB development if no-primary sections acquire color; these are general chromogenic workflow checks (standard IHC practice). Treat signal in adipocytes or bone-marrow hematopoietic cells cautiously because these populations are reported as not detected, while kidney collecting ducts are reported high (HPA tissue IHC).
What is a defensible way to quantify HSD11B2 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and cell population before scoring, such as kidney collecting ducts or placental syncytiotrophoblast cell bodies, both reported as high-staining sites (HPA tissue IHC). Record the percentage of positive target cells and staining intensity, then calculate an H-score using intensity categories 0–3 and their corresponding percentages (standard IHC practice). Keep retrieval, the catalog antibody's 1 μg/ml concentration, DAB development, illumination, and image-analysis thresholds consistent across the comparison set (datasheet RP1094; standard IHC practice). Normalize positive-cell counts to the number of eligible target cells, or stained area to sampled compartment area, and report excluded folds and necrotic regions (standard IHC practice).
When should apparent HSD11B2 positivity be considered an artefact?
A credible result places cytoplasmic stain in an expected population, such as kidney collecting ducts, alongside a suitable same-run positive control (HPA tissue IHC; standard IHC practice). Question predominantly nuclear color, extracellular precipitate, and strong staining confined to tissue edges or necrotic areas, because these patterns do not match the reported tissue profile or support a reliable cellular assignment (HPA tissue IHC; standard IHC practice). Compare any suspicious deposit with a no-primary section and verify that endogenous peroxidase was blocked before DAB development (standard IHC practice). Interpret staining intensity as protein detection, not as a direct measure of cortisol-to-cortisone conversion or enzyme activity (UniProt P80365; standard IHC practice).
Boster reagents
Best HSD11B2 / 11-beta-hydroxysteroid dehydrogenase type 2 IHC Antibodies
Anti-HSD11B2 antibodies have IHC images from human placenta, colon, cancers, and mouse and rat pancreas; both have IF images from human placenta (catalog image captions).
Real IHC data
Anti-HSD11B2 Antibody ®
Cat # RP1094
Real IHC data
Anti-HSD11B2 Antibody ®
Cat # A01613
RP1094 is listed for human, mouse, and rat IHC and IF, with paraffin-section IHC images from human placenta and mouse and rat pancreas (RP1094 applications, reactivity, and IHC captions). A01613 is listed for human IHC and IF, with paraffin-section IHC images from human colon, appendix cancer, breast cancer, and endometrial cancer (A01613 applications, reactivity, and IHC captions).
Which to pick: For human tissue IHC, choose A01613 when its human paraffin-section examples fit the sample; its listed IHC concentration is 2–5 μg/ml (A01613 IHC captions; catalog dilution). Choose RP1094 for cross-species tissue IHC: its paraffin-section captions show human, mouse, and rat samples, and its listed IHC concentration is 0.5–1 μg/ml (RP1094 IHC captions; catalog dilution). For IF, both have human placenta section images at 5 μg/ml; ICC validation and fixation method are unreported in these captions (A01613 and RP1094 IF and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P80365 (DHI2_HUMAN, 11-beta-hydroxysteroid dehydrogenase type 2).
- Human Protein Atlas. HSD11B2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HSD11B2 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. HSD11B2 antibody validation summary (3 antibodies).
- Primary Uterine Inertia (PUI) in Dogs Is Associated with Impaired Placental Availability of Factors Involved in the Parturition Cascade. Animals : an open access journal from MDPI 2025 — PMC12561314.
- Identification and clinical validation of key genes as the potential biomarkers in colorectal adenoma. BMC cancer 2023 — PMC9832797.
- Primary aldosteronism patients show skin alterations and abnormal activation of glucocorticoid receptor in keratinocytes. Scientific reports 2017 — PMC5693903.
- 11β-Hydroxysteroid dehydrogenase type 2 may mediate the stress-specific effects of cortisol on brain cell proliferation in adult zebrafish (Danio rerio). The Journal of experimental biology 2024 — PMC11418181.
- PubMed PMID:7859916 — UniProt-cited evidence.
- PubMed PMID:8530071 — UniProt-cited evidence.
- PubMed PMID:8611140 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSD11B2 (11-beta-hydroxysteroid dehydrogenase type 2). Expected localisation Cytoplasmic staining in select tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet RP1094). Validated IHC antibody: RP1094 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Mouse Pancreas tissue; fixative not specified (datasheet RP1094); 2. Fixation: Image fixative and duration unreported (datasheet RP1094); 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 RP1094); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet RP1094); 8. Primary antibody: Rabbit anti-HSD11B2, 0.5-1μg/ml (datasheet RP1094); 9. Primary incubation: Overnight at 4 °C (datasheet RP1094). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image RP1094); Caveat: Restricted positive cell types can be missed in bulk scoring (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one 1–405 chain (UniProt); Regulation: Tissue-dependent expression (UniProt). Evidence: UniProt P80365, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet RP1094, PMC12561314, PMC9832797. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSD11B2/immunohistochemistry-ihc-guide.