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HMOX2 / Heme oxygenase 2 · IHC design guide
Design Immunohistochemistry for HMOX2
Plan chromogenic HMOX2 IHC on paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet: PB9213). Compare cytoplasmic staining in high-expression appendix glandular cells with low-expression parathyroid glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HMOX2 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 (HPA tissue IHC); ER membrane expected (UniProt) | |
| Staining pattern | Widespread cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9213) | |
| Positive control | Appendix+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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9213) | |
| Caveat | Parathyroid glandular cells may show weak staining (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | Two isoforms; soluble 2–295 lacks C-terminal epitopes (UniProt) |
Section 1
Recommended HMOX2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PB9213) is accompanied by one published HMOX2 chromogenic IHC protocol (PMC13013296).
| Sample | Paraffin-embedded human appendix mucinous adenocarcinoma tissue; fixative not specified (datasheet PB9213) |
| Fixation | Image fixative and duration unreported (datasheet PB9213); 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 PB9213); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9213) |
| Primary antibody | Rabbit anti-HMOX2, 2-5μg/ml (datasheet PB9213) |
| Primary incubation | Overnight at 4 °C (datasheet PB9213) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9213) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HMOX2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: PB9213). The published protocol does not specify antigen retrieval (PMC13013296).
Section 2
What Is the Expected HMOX2 Staining Pattern?
HMOX2 should appear predominantly in the cytoplasm of many cell types, including glandular cells in appendix and colon and neuronal cells in caudate (HPA: ubiquitous cytoplasmic expression; High in these cells). Its cytoplasmic region spans residues 2–295, with a membrane anchor at 296–316 (UniProt P30519 topology). HPA rates the tissue staining pattern Supported because antibody staining is consistent with RNA expression (HPA: reliability Supported).
| Clear cytoplasmic staining in appendix or colon glandular cells, with tissue structures still discernible. | This matches cells scored High in those tissues and the reported ubiquitous cytoplasmic profile (HPA: appendix and colon glandular cells High; ubiquitous cytoplasmic expression). Compare cell-associated signal with a matched control slide before interpreting intensity (standard IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic signal in cells expected to stain. | A nuclear-dominant pattern conflicts with the reported cytoplasmic tissue profile and the cytoplasmic orientation of most of HMOX2 (HPA: ubiquitous cytoplasmic expression; UniProt P30519 topology). Treat it as suspect and check the detection controls before assigning it to HMOX2 (standard IHC practice). |
| Strong staining concentrated in cardiomyocytes or adipocytes while an expected positive tissue is weak. | Those cells are scored Low, so the distribution warrants a cross-reactivity or endogenous detection activity check (HPA: cardiomyocytes Low; adipocytes Low; standard IHC practice). Low means weak reported staining, not a proven absence of HMOX2 (HPA: Low). |
| Uniform color across cells, stroma, and empty spaces, with indistinct cell boundaries. | That distribution is less persuasive than cell-associated cytoplasmic staining (HPA: ubiquitous cytoplasmic expression). Diffuse background can arise from detection or blocking conditions; inspect a no-primary control and the section for nonspecific precipitate (standard IHC practice). |
| No convincing cell-associated signal in appendix or colon glandular cells. | An absent result conflicts with their reported High staining, but does not by itself establish that the tissue lacks HMOX2 (HPA: appendix and colon glandular cells High). Review section integrity, retrieval, antibody incubation, and detection with a known-positive control (standard IHC practice). |
💡Expected HMOX2 appearanceCall the IHC result positive when discernible cytoplasmic signal appears in expected cells, especially strongly stained appendix or colon glandular cells; nuclear-dominant staining or uniform background is suspect (HPA: ubiquitous cytoplasmic expression; appendix and colon glandular cells High; UniProt P30519 topology; standard IHC practice).
| Membrane topology (UniProt P30519 topology) | HMOX2 is assigned to microsome and endoplasmic reticulum membranes, with residues 2–295 cytoplasmic and 296–316 transmembrane (UniProt P30519 topology). Interpret a cytoplasmic IHC profile in that context; the record does not define a required punctate appearance in paraffin sections (HPA: ubiquitous cytoplasmic expression). |
| Processed form and isoforms (UniProt P30519 processing; isoforms) | The record lists membrane-associated chain 2–316, soluble chain 2–295, and 2 isoforms (UniProt P30519 processing; isoforms). Without an antibody epitope in the supplied evidence, a staining pattern cannot distinguish those forms (UniProt P30519 processing; standard IHC interpretation). |
| Evidence strength across tissues (HPA: reliability Supported; tissue IHC) | HPA describes the tissue profile as Supported and reports High staining in several glandular and neuronal populations, alongside Low staining in other cells (HPA: reliability Supported; tissue IHC). Use those levels as comparison points, not as universal cutoffs for slide scoring (standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive control is blank. | The slide offers no evidence that the IHC run detected an expected High population (HPA: appendix and colon glandular cells High). | Check control tissue quality and each retrieval, primary-antibody, and detection step; repeat with a known-positive section before interpreting experimental negatives (standard IHC practice). |
| Signal is mainly nuclear. | Nuclear dominance disagrees with the HPA cytoplasmic profile and UniProt membrane topology (HPA: ubiquitous cytoplasmic expression; UniProt P30519 topology). | Review a no-primary control, counterstain, and chromogen distribution; confirm cell-associated cytoplasmic signal in a known-positive tissue (standard IHC practice; HPA: appendix glandular cells High). |
| Color covers the whole section, including spaces between cells. | A spatially uniform deposit may reflect nonspecific detection or chromogen background rather than localized staining (standard IHC practice). | Compare a no-primary control, review blocking and detection conditions, and score only interpretable cell-associated staining (standard IHC practice; HPA: ubiquitous cytoplasmic expression). |
| Low-scored cells stain more strongly than expected positive cells. | A reversed distribution raises concern for cross-reactivity or endogenous detection activity; Low is not a negative designation (HPA: cardiomyocytes Low; appendix glandular cells High; standard IHC practice). | Compare tissue controls and a no-primary control, then assess whether the signal follows cell type and cytoplasmic location (standard IHC practice; HPA: ubiquitous cytoplasmic expression). |
| A faint cytoplasmic signal is difficult to score. | HPA reports both High and Low staining across cell populations, so weak color alone cannot identify a failed run (HPA: tissue IHC). | Judge signal against matched control tissue, background, and preserved morphology; record intensity and the stained cell type separately (standard IHC practice). |
| What pattern should be expected in IF/ICC? | The IF/ICC evidence describes vesicles and cytosol, a readout distinct from chromogenic tissue IHC (HPA: subcellular ICC-IF; tissue IHC). | Look for vesicular and cytosolic localization in the IF/ICC guide; HPA marks both locations Approved and lists A-431, U-251MG, and U2OS images (HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing HMOX2 staining
PB9213 · IHCIHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213). HMOX2 was detected in a paraffin-embedded section of human appendix mucinous adeno
PB9213 · IHCIHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213). HMOX2 was detected in a paraffin-embedded section of human colorectal adenocarcino
A05506-2 · IHCIHC analysis of Heme oxygenase 2/HMOX2 using anti-Heme oxygenase 2/HMOX2 antibody (A05506-2). Heme oxygenase 2/HMOX2 was detected in a paraf
A05506-2 · IHCIHC analysis of Heme oxygenase 2/HMOX2 using anti-Heme oxygenase 2/HMOX2 antibody (A05506-2). Heme oxygenase 2/HMOX2 was detected in a paraf
A05506-2 · IFIF analysis of Heme oxygenase 2/HMOX2 using anti-Heme oxygenase 2/HMOX2 antibody (A05506-2). Heme oxygenase 2/HMOX2 was detected in an immun
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000103415-HMOX2/tissue/cerebral+cortex#imid_11090751
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000103415-HMOX2/tissue/colon#imid_11096501
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000103415-HMOX2/tissue/liver#imid_11096502
HPA · GallbladderHuman Protein Atlas IHC, Gallbladder. Source: https://www.proteinatlas.org/ENSG00000103415-HMOX2/tissue/gallbladder#imid_11096494
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB025464; green = HMOX2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB025464; green = HMOX2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HMOX2 IHC & IF
🧪Run appendix first and look for HMOX2 staining in its glandular cells (HPA: High in appendix glandular cells). HPA detects HMOX2 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells without specific staining on the appendix slide as background without assuming a named cell type is HMOX2-negative (HPA: no negative rows; detected in all 45 tissues).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: HMOX2 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 HMOX2 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control, and HMOX2-knockout material if available as a biological negative (selected caption: rabbit primary; standard IHC specificity practice). Quench endogenous peroxidase and check for background in inflammatory cells when interpreting appendix HRP/DAB staining (selected caption: appendix HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9213 paraffin-section caption does not state the fixative (selected caption: fixative not stated). The reported IHC example uses heat-mediated EDTA retrieval at pH 8.0; whether HMOX2 staining depends on that retrieval condition is unreported (selected caption: EDTA pH 8.0 retrieval). Frozen-section performance and whether tissue IF is easier are unreported; ICC-IF images show vesicular and cytosolic localisation in A-431, U-251MG and U2OS, while appendix inflammatory cells can contribute endogenous peroxidase background in HRP/DAB IHC (HPA: subcellular ICC-IF; selected caption: appendix HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for HMOX2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: HMOX2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced HMOX2 IHC Tips
Troubleshoot HMOX2 staining in paraffin sections using the catalog antibody’s tissue IHC conditions and compartment specific controls.
Which retrieval conditions should I start with for HMOX2 paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet PB9213). Apply the same heating and cooling schedule across sections, then compare staining in glandular cells of appendix, where HMOX2 is reported as high, with a no primary control (HPA tissue IHC; standard IHC practice). The selected image used 2 μg/ml antibody overnight at 4°C after retrieval, so keep those conditions fixed while assessing retrieval (datasheet PB9213). If staining remains weak, compare a second retrieval condition on adjacent sections and judge both signal and tissue preservation before changing the antibody concentration (standard IHC practice).
How should I troubleshoot fixation when HMOX2 staining varies between paraffin blocks?
The selected PB9213 caption identifies a paraffin section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet PB9213). Record each block’s documented fixation conditions and compare sections of similar thickness and processing history before assigning the difference to HMOX2 abundance (standard IHC practice). Use the same EDTA pH 8.0 retrieval and antibody incubation across comparison sections, then inspect nuclear detail and tissue integrity alongside staining (datasheet PB9213; standard IHC practice). If fixation records differ, report that limitation and repeat the comparison in consistently processed material when available; neither topology nor tissue staining establishes a fixation effect (standard IHC practice).
What HMOX2 staining pattern should I expect in chromogenic tissue IHC?
Look for predominantly cytoplasmic staining, with a possible fine membranous distribution: HMOX2 is associated with microsome and endoplasmic reticulum membranes, and residues 2–295 face the cytoplasm (HPA tissue IHC; UniProt P30519 topology). HPA describes ubiquitous cytoplasmic tissue expression and high staining in appendix glandular cells, which provides a useful cell level comparison for the selected paraffin image (HPA tissue IHC; datasheet PB9213). Review staining at high magnification against cell borders and nuclei rather than treating all brown deposits as intracellular signal (standard IHC practice). Predominantly nuclear staining would require independent validation because the supplied localisation evidence does not support that assignment (UniProt P30519; HPA tissue IHC).
Could HMOX2 isoforms or epitope placement explain discordant IHC staining?
Check the antibody’s disclosed immunogen or epitope before interpreting discordance: the supplied record lists 2 isoforms but does not map this antibody’s binding site (UniProt P30519; datasheet PB9213). The annotated membrane segment spans residues 296–316, while residues 2–295 are cytoplasmic and a soluble 2–295 chain is listed (UniProt P30519 topology and processing). Thus, epitope placement could affect which forms are detected, but the present evidence cannot establish that PB9213 distinguishes them (UniProt P30519; datasheet PB9213). Compare adjacent sections under identical retrieval and detection conditions, and describe staining by cell and compartment until epitope specificity is documented (standard IHC practice).
How can I assess HMOX2 by multiplex IF alongside this chromogenic IHC guide?
Treat IF as a separate assay requiring its own antibody validation; the PB9213 paraffin IHC caption does not establish IF performance (datasheet PB9213; standard IF practice). Pair HMOX2 with a validated glandular cell marker in appendix and include single color controls to assess channel bleed through (HPA tissue IHC; standard IF practice). Choose fluorophores and exposure settings after checking the tissue’s autofluorescence, favoring a spectrally separated channel where background is lowest (standard IF practice). Because residues 2–295 face the cytoplasm, optimize permeabilisation for access to any epitope on that side, while checking morphology and background against an unpermeabilised control (UniProt P30519 topology; standard IF practice).
How do I distinguish diffuse HMOX2 staining from chromogenic background?
Run a no primary control through the same secondary, peroxidase block and DAB development to reveal detection background (standard IHC practice). The selected image used 10% goat serum block, 2 μg/ml primary overnight at 4°C, and a 30 minute secondary incubation at 37°C (datasheet PB9213). If both test and control sections stain diffusely, review peroxidase blocking, wash steps and DAB development time before reducing primary concentration (standard IHC practice). Expect some widespread cytoplasmic signal because HPA describes ubiquitous expression; evaluate whether deposits remain cell associated and exceed the matched control (HPA tissue IHC; standard IHC practice).
How should I score HMOX2 staining across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring; HPA reports high HMOX2 in appendix glandular cells and a broadly cytoplasmic tissue profile (HPA tissue IHC). For those cells, record percentage positive and intensity on a fixed 0–3 scale, then calculate an H score as the sum of intensity multiplied by percentage at each intensity (standard IHC practice). Normalize counts to eligible viable cells, or stained area to analyzed viable tissue area in mm², using the same threshold across slides (standard IHC practice). Exclude folds, necrosis and section edges by predefined rules, and compare batches only after checking matched controls and staining settings (standard IHC practice).
When is an apparent HMOX2 positive result likely to be artefactual?
A convincing result shows cell associated cytoplasmic signal in an expected population, such as appendix glandular cells, above the no primary control (HPA tissue IHC; standard IHC practice). HMOX2 is linked to endoplasmic reticulum and microsome membranes, with a cytoplasmic domain, so isolated nuclear DAB deposits warrant scrutiny (UniProt P30519 topology). Discount staining confined to section edges, folds or necrotic areas, and investigate diffuse deposits that persist without primary antibody as potential detection artefacts (standard IHC practice). Review peroxidase blocking and matched DAB development when suspected endogenous enzyme activity is present; confirm uncertain patterns with an independently validated assay before assigning biological meaning (standard IHC practice).
Boster reagents
Best HMOX2 / Heme oxygenase 2 IHC Antibodies
Both antibodies have human paraffin-section IHC images (PB9213 and A05506-2 IHC captions); A05506-2 also has an IF image of MCF-7 cells (A05506-2 IF caption).
Real IHC data
Anti-Heme oxygenase 2/HMOX2 Antibody ®
Cat # PB9213
Real IHC data
Anti-Heme oxygenase 2/HMOX2 Antibody ®
Cat # A05506-2
PB9213 will render with a human appendix mucinous adenocarcinoma paraffin-section IHC image; its listed reactivity is human, mouse and rat (PB9213 IHC caption; catalog reactivity). A05506-2 will render with a human lung squamous cell carcinoma paraffin-section IHC image and is also listed for human IF/ICC, with an IF image of MCF-7 cells (A05506-2 IHC and IF captions; catalog applications and reactivity).
Which to pick: For human tissue IHC, either SKU has its own paraffin-section image using EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; fixation is unreported for both (PB9213 and A05506-2 IHC captions). Choose A05506-2 for human IF/ICC because IF and ICC are listed applications and its MCF-7 IF caption documents 5 μg/ml primary antibody (A05506-2 catalog applications and IF caption). Choose PB9213 when mouse or rat reactivity is needed, while treating its pictured IHC evidence as human only; clonality is unreported for both SKUs (PB9213 catalog reactivity and IHC caption; catalog clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30519 (HMOX2_HUMAN, Heme oxygenase 2).
- Human Protein Atlas. HMOX2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HMOX2 subcellular location (ICC-IF): Localized to vesicles and cytosol..
- Human Protein Atlas. HMOX2 antibody validation summary (2 antibodies).
- Iron toxicity undermines microfracture-induced cartilage regeneration by predisposing a pre-ferroptotic niche. Frontiers in cell and developmental biology 2026 — PMC13013296.
- PubMed PMID:7890772 — UniProt-cited evidence.
- PubMed PMID:1575508 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HMOX2 (Heme oxygenase 2). Expected localisation Cytoplasmic staining (HPA tissue IHC); ER membrane expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9213). Validated IHC antibody: PB9213 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (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 appendix mucinous adenocarcinoma tissue; fixative not specified (datasheet PB9213); 2. Fixation: Image fixative and duration unreported (datasheet PB9213); 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 PB9213); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9213); 8. Primary antibody: Rabbit anti-HMOX2, 2-5μg/ml (datasheet PB9213); 9. Primary incubation: Overnight at 4 °C (datasheet PB9213). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9213); Caveat: Parathyroid glandular cells may show weak staining (HPA tissue IHC); Isoform / epitope: Two isoforms; soluble 2–295 lacks C-terminal epitopes (UniProt); Regulation: Expression regulation is not specified (UniProt). Evidence: UniProt P30519, HPA tissue IHC · Supported, HPA ICC-IF · vesicles and cytosol., Datasheet PB9213, PMC13013296. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HMOX2/immunohistochemistry-ihc-guide.