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OXR1 / Oxidation resistance protein 1 · IHC design guide
Design Immunohistochemistry for OXR1
Use the cytoplasmic tissue pattern and high-staining cell types to plan OXR1 paraffin-section IHC (HPA tissue IHC). Start antibody titration within 1:100–1:300 and compare staining across cell types (datasheet: 1:100–1:300; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
OXR1 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 across tissues (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Ovary+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Staining levels vary across tissues (HPA tissue IHC) | |
| Isoform / epitope | 8 isoforms; epitope coverage depends on sequence (UniProt) |
Section 1
Recommended OXR1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by one published OXR1 kidney IHC protocol (PMC4435960).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A04583-1) |
| Fixation | Image fixative and duration unreported (datasheet A04583-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-OXR1, 1:100-1:300 (datasheet A04583-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | OXR1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues at variable levels. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER, 95–98 °C for 20 min (page retrieval rule); the kidney article does not report retrieval (PMC4435960).
Section 2
What Is the Expected OXR1 Staining Pattern?
OXR1 should appear mainly as variable cytoplasmic staining in tissue IHC, with strong signal in selected glandular, neuronal, and germ-cell populations (HPA: tissue IHC, Enhanced reliability). Its reported locations include mitochondria, the nucleolus, and Golgi-associated membranes (UniProt Q8N573: subcellular location). OXR1 has no transmembrane segment, so a continuous cell-surface rim is unexpected (UniProt Q8N573: topology).
| Cytoplasmic signal is strong in adrenal or endometrial glandular cells, cortical neurons, or testicular pachytene spermatocytes (HPA: High in each). | This fits the observed tissue pattern. Compare the named cells with adjacent structures on the same section; OXR1 intensity varies by tissue, so equal staining across all cells is unnecessary (HPA: variable cytoplasmic expression). |
| Signal is predominantly a continuous cell-surface rim, with little cytoplasmic staining. | Check for an artefact or off-target staining: OXR1 lacks a transmembrane segment (UniProt Q8N573: topology), while tissue IHC is chiefly cytoplasmic (HPA: tissue IHC). Discrete organelles may be difficult to resolve by chromogenic IHC (general IHC practice). |
| Strong staining appears in ovarian stromal cells, skin keratinocytes, or vaginal squamous epithelium (HPA: Not detected in each). | Treat this as unexpected for the named cell population, rather than proof of cross-reactivity. Review morphology and controls for off-target binding or endogenous detection activity (HPA: tissue IHC; general IHC practice). |
| Color is spread across the section or tissue-free areas, obscuring cell boundaries. | This is background rather than a readable OXR1 cell pattern (general IHC practice). Check reagent-only controls, blocking, washes, and detection conditions before assigning intensity scores (general IHC practice). |
| No signal appears in a well-preserved, expected-positive cell population. | A negative result is inconclusive if the run fails in a High group, such as cerebellar granular cells or salivary glandular cells (HPA: High). Check the positive control and assay conditions before scoring the test section negative (general IHC practice). |
💡Expected OXR1 appearanceA convincing positive is cell-associated cytoplasmic staining, potentially High in HPA-listed glandular or neuronal cells; a continuous surface rim or broad section-wide color warrants review (HPA: tissue IHC; UniProt Q8N573: topology; general IHC practice).
| Cell population and tissue (HPA: tissue IHC) | HPA reports High staining in adrenal, endometrial, and salivary glandular cells, cerebral cortical neurons, cerebellar granular cells, and pachytene spermatocytes; adipocytes are Medium. Use the specific cell population, rather than the organ alone, when comparing slides (HPA: tissue IHC). |
| Compartment and resolution (HPA: tissue IHC; UniProt Q8N573: subcellular location) | Tissue IHC is described as cytoplasmic. UniProt reports mitochondria in neuronal cells, nucleoli in fibroblasts and myoblasts, and Rab-dependent recruitment to Golgi membranes; these finer assignments should not be required from routine chromogenic sections (HPA: tissue IHC; UniProt Q8N573: subcellular location; general IHC practice). |
| Strength of tissue evidence (HPA: Enhanced reliability) | HPA labels tissue staining Enhanced, yet describes only medium consistency between staining and RNA data. Its three listed antibodies have Enhanced IHC status; that supports the reported pattern without making any single positive or negative cell an absolute diagnostic rule (HPA: tissue IHC; HPA: HPA027375, HPA027380, HPA027395). |
| Isoforms and epitope coverage (UniProt Q8N573: isoforms) | UniProt lists 8 isoforms. Whether a particular IHC antibody detects each one depends on its epitope, which is not supplied here; do not infer an isoform-specific tissue pattern or use one antibody's negative result to exclude every isoform (UniProt Q8N573: isoforms; general immunostaining practice). |
| IF/ICC expectation? (HPA: subcellular ICC-IF) | HPA's approved main ICC-IF location is vesicles, with images in U2OS and A-431. That is useful context for a separate IF/ICC design, while tissue IHC should be read against its cytoplasmic cell pattern; two listed antibodies have Uncertain ICC status (HPA: subcellular ICC-IF; HPA: HPA027380, HPA027395). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control tissue is blank. | The run may lack detectable signal; absence in a High population is unexpected (HPA: tissue IHC). | Verify tissue identity and the IHC-validated antibody's documented conditions; inspect detection and counterstain controls before interpreting test negatives (general IHC practice). |
| Test section is blank but the positive control stains. | The sampled cells may express less OXR1: HPA reports variable tissue levels and some cell populations as Not detected (HPA: tissue IHC). | Identify the actual cell population and compare it with HPA's cell-specific entries; record a negative only for the sampled cells under the validated run conditions (HPA: tissue IHC; general IHC practice). |
| Strong color appears in tissue-free areas or across many unrelated structures. | Diffuse deposition can arise from nonspecific detection or inadequate washing (general IHC practice). | Review the reagent-only control, blocking, washes, and detection time; score cells only once background permits reliable boundaries (general IHC practice). |
| Unexpected cell-surface rims dominate the section. | That distribution conflicts with predominantly cytoplasmic tissue staining and a protein without a transmembrane segment (HPA: tissue IHC; UniProt Q8N573: topology). | Review morphology and controls, then compare with staining from another IHC-validated antibody if available; do not call the rim specific from color alone (HPA: antibody IHC status; general IHC practice). |
| HPA-listed Not detected cells stain strongly. | Off-target binding or endogenous detection activity is possible; HPA's absence call is cell-specific rather than an organ-wide rule (HPA: tissue IHC; general IHC practice). | Confirm cell identity, inspect a reagent-only control, and assess endogenous detection activity appropriate to the chromogenic system before interpreting OXR1 positivity (general IHC practice). |
| Fine puncta or nucleolar color are inconsistent between sections. | UniProt reports several context-dependent locations, while chromogenic section resolution and tissue morphology can limit compartment assignment (UniProt Q8N573: subcellular location; general IHC practice). | Prioritize reproducible, cell-associated cytoplasmic staining in expected populations; use the separate IF/ICC guide for finer localisation questions (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing OXR1 staining
A04583-1 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using OXR1 Antibody. The picture on the right is blocked with the syn
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164830-OXR1/tissue/cerebral+cortex#imid_6845752
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164830-OXR1/tissue/colon#imid_6829254
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164830-OXR1/tissue/liver#imid_6829263
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164830-OXR1/tissue/kidney#imid_6829248
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA027380; green = OXR1, blue = nucleus, red = microtubules.
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA027380; green = OXR1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for OXR1 IHC & IF
🧪Run cerebellum first; granular cells should stain (High; HPA: cerebellum granular cells). Run skin as the negative tissue; keratinocytes are Not detected (HPA: skin keratinocytes). On the cerebellar slide, use nongranular cells as background comparators, without assuming they are OXR1-negative (HPA: High is assigned to granular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Ovary (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show OXR1 in U2OS, A-431, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host species and clonality, and knockout tissue if available; the peptide-blocked human brain section provides a separate competition control (selected-SKU IHC caption: A04583-1). Quench endogenous peroxidase for chromogenic detection and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: An OXR1-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval against the positive and negative controls (standard IHC practice). The paraffin-section human brain caption does not establish fixation, and its fixative is unreported (selected-SKU IHC caption: A04583-1); the supplied evidence does not establish whether frozen sections or IF are easier. Cerebellar lipofuscin may cause autofluorescent background in IF, so check an unstained or no-primary section (standard IF practice). The selected A04583-1 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A04583-1).
HPA tissue IHC evidence for OXR1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 → |
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced OXR1 IHC Tips
Troubleshoot OXR1 staining in paraffin sections by checking retrieval, cell type, compartment, and controls before interpreting chromogenic signal.
How should I retrieve OXR1 in paraffin sections when staining is weak?
Use Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min before chromogenic IHC (page antigen retrieval setting). Keep section thickness, cooling time, and detection conditions constant while comparing a short retrieval time series, because excessive heating can damage tissue morphology (standard IHC practice). Evaluate signal in cerebral cortex neuronal cells or cerebellar granular cells, both reported as high-staining populations, alongside an unstained control section (HPA: tissue IHC). If staining remains weak, test citrate pH 6.0 as a fallback on serial sections and record the effect on signal and background (standard IHC practice).
Could fixation explain inconsistent OXR1 staining between paraffin blocks?
The selected image documents OXR1 staining in paraffin-embedded human brain, but its caption gives no fixative; OXR1-specific fixation sensitivity is therefore unknown (A04583-1 caption). Record each block’s fixative, fixation duration, processing history, and section age before attributing a difference to OXR1 expression (standard IHC practice). Compare serial sections with the same Tris-EDTA pH 9.0, 95–98 °C, 20 min retrieval and identical chromogenic detection (page antigen retrieval setting). Include a known staining region from a reference block in each run, and judge both cell morphology and signal before changing fixation or retrieval conditions (standard IHC practice).
Which OXR1 staining patterns are plausible in tissue sections?
Expect predominantly cytoplasmic staining across tissues at variable levels, with high staining reported in cerebral cortex neuronal cells and cerebellar granular cells (HPA: tissue IHC). Vesicles are the approved main location in cell imaging, while mitochondria in neuronal cells, nucleoli in fibroblasts and myoblasts, and Rab-dependent Golgi recruitment are also reported (HPA: subcellular; UniProt Q8N573). In chromogenic sections, compare the apparent compartment within the expected cell population against adjacent cells and a counterstained serial section (standard IHC practice). Treat exclusively nuclear, diffuse extracellular, or uniformly membrane-rim staining as requiring additional controls before assigning it to OXR1 (UniProt Q8N573 localisation; standard IHC practice).
How can isoforms or epitope location affect OXR1 IHC interpretation?
OXR1 has 8 annotated isoforms and LysM, GRAM, and TLDc domains at residues 99–142, 208–275, and 713–874, respectively (UniProt Q8N573). Determine which sequence the antibody recognizes before assuming that every OXR1 isoform contributes equally to a tissue signal (UniProt Q8N573 isoforms; standard IHC practice). The selected brain image includes peptide blocking, but its caption does not map the epitope or establish isoform coverage (A04583-1 caption). If staining differs between cell populations, compare an independent antibody with a mapped, distinct epitope and document retrieval conditions before interpreting the difference biologically (standard IHC practice).
How should I assess OXR1 by IF alongside the chromogenic IHC result?
For IF on a matching brain section, multiplex OXR1 with a neuronal marker to check whether signal falls in the expected cerebral cortex neuronal population (HPA: tissue IHC). Choose a spectrally separated, preferably far-red fluorophore and inspect an unstained section for tissue autofluorescence before assigning puncta to OXR1 (standard IF practice). Because OXR1 has no transmembrane segment and can associate with Rab-dependent Golgi membranes, optimize mild permeabilisation for access to the antibody’s mapped epitope without assuming its orientation (UniProt Q8N573 topology and localisation; standard IF practice). Compare the IF compartment pattern with chromogenic sections and include single-label and no-primary controls for bleed-through and background (standard IF practice).
What controls help separate OXR1 signal from chromogenic background?
Run a no-primary control and block endogenous peroxidase before DAB development to identify detection-system staining that can mimic OXR1 signal (standard chromogenic IHC practice). The selected human brain image includes a synthesized-peptide blocking comparison, which can assess competition for this antibody but cannot alone prove tissue specificity (A04583-1 caption; standard IHC practice). Check high-staining cerebral cortex neuronal cells against keratinocytes, reported as not detected, while allowing for differences between tissues and runs (HPA: tissue IHC). If background persists, titrate primary antibody and detection time on serial sections, preserving the same retrieval and counterstain conditions for comparison (standard IHC practice).
How should I score OXR1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports variable cytoplasmic OXR1 staining across tissues and high staining in selected neuronal and glandular populations (HPA: tissue IHC). For comparable chromogenic sections, score the percentage of positive target cells and intensity on a 0–3 scale, then calculate an H-score from 0–300 (standard IHC practice). Normalize positive-cell counts to the number of intact target cells, or stained-cell density to analyzed tissue area in mm², rather than whole-section area (standard IHC practice). Exclude folds, edges, and necrotic areas using predefined criteria, and keep retrieval, exposure, and threshold settings consistent between batches (standard IHC practice).
When should an apparent OXR1-positive region be considered artefactual?
A credible OXR1 result should track an appropriate cell population and predominantly cytoplasmic pattern, such as reported cerebral cortex neuronal staining (HPA: tissue IHC). Punctate signal can be plausible given the approved vesicular location and reported neuronal mitochondrial or Rab-dependent Golgi association, but chromogenic puncta alone cannot identify an organelle (HPA: subcellular; UniProt Q8N573). Investigate staining confined to section edges, necrotic tissue, or no-primary controls as possible processing or endogenous-enzyme artefact (standard IHC practice). Compare serial sections, morphology, and the selected antibody’s peptide-blocking result before treating unexpected nuclear-only or extracellular staining as evidence of OXR1 localisation (A04583-1 caption; standard IHC practice).
Boster reagents
Best OXR1 / Oxidation resistance protein 1 IHC Antibodies
A04583-1 has a human brain paraffin-section IHC image (IHC caption), listed IF use, and Human, Mouse, and Rat reactivity (catalog applications and reactivity).
Real IHC data
Anti-Oxidation resistance protein 1 Oxr1 Antibody
Cat # A04583-1
A04583-1 has an IHC image of paraffin-embedded human brain tissue with a peptide-blocked comparison (IHC caption). IF is listed as an application, and Human, Mouse, and Rat are listed as reactive species; no IF image is supplied (catalog applications, reactivity, and image records).
Which to pick: For tissue IHC, choose A04583-1: its own image shows staining in paraffin-embedded human brain tissue with a peptide-blocked comparison; the fixative is unreported (IHC caption). For IF, A04583-1 is listed at 1:50, but ICC is not separately listed and no IF image is supplied (catalog applications, dilution, and image records). For Human, Mouse, or Rat samples, A04583-1 lists reactivity across those species; the supplied IHC image documents Human tissue only, and clonality is unreported (catalog reactivity, IHC caption, and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N573 (OXR1_HUMAN, Oxidation resistance protein 1).
- Human Protein Atlas. OXR1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. OXR1 subcellular location (ICC-IF): Mainly localized to vesicles..
- Human Protein Atlas. OXR1 antibody validation summary (3 antibodies).
- Knockdown of orexin type 1 receptor in rat locus coeruleus increases REM sleep during the dark period. The European journal of neuroscience 2010 — PMC3058252.
- Delivering Oxidation Resistance-1 (OXR1) to Mouse Kidney by Genetic Modified Mesenchymal Stem Cells Exhibited Enhanced Protection against Nephrotoxic Serum Induced Renal Injury and Lupus Nephritis. Journal of stem cell research & therapy 2014 — PMC4435960.
- Molecular Assessment of Epiretinal Membrane: Activated Microglia, Oxidative Stress and Inflammation. Antioxidants (Basel, Switzerland) 2020 — PMC7465764.
- Iridium metal complex targeting oxidation resistance 1 protein attenuates spinal cord injury by inhibiting oxidative stress-associated reactive oxygen species. Redox biology 2023 — PMC10587759.
- PubMed PMID:12880961 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for OXR1 (Oxidation resistance protein 1). Expected localisation Cytoplasmic staining across tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A04583-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Ovary. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A04583-1); 2. Fixation: Image fixative and duration unreported (datasheet A04583-1); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-OXR1, 1:100-1:300 (datasheet A04583-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 8 isoforms; epitope coverage depends on sequence (UniProt); Regulation: Staining levels vary across tissues (HPA tissue IHC). Evidence: UniProt Q8N573, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to vesicles., Datasheet A04583-1, PMC4435960. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/OXR1/immunohistochemistry-ihc-guide.