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OCRL / Inositol polyphosphate 5-phosphatase OCRL · IHC design guide
Design Immunohistochemistry for OCRL
Plan chromogenic OCRL IHC on paraffin sections with 2–5 μg/ml of the IHC-validated antibody (datasheet A02042-2). Assess cytoplasmic staining in glandular and neuronal cells, where HPA reports high signal in several tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
OCRL 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in glandular and neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02042-2) | |
| 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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02042-2) | |
| Caveat | Staining and RNA levels show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms (A/B); epitope coverage unknown (UniProt) |
Section 1
Recommended OCRL IHC & IF Protocols
The catalog antibody protocol uses paraffin sections; one published article provides additional brain-section preparation and DAB staining details for IHC (PMC12192959).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A02042-2) |
| Fixation | Image fixative and duration unreported (datasheet A02042-2); 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 A02042-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02042-2) |
| Primary antibody | Rabbit anti-OCRL, 2-5 μg/ml (datasheet A02042-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02042-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02042-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | OCRL-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A02042-2); the published excerpt does not specify retrieval (PMC12192959).
Section 2
What Is the Expected OCRL Staining Pattern?
OCRL is a cytoplasmic, membrane-associated protein with no transmembrane segment; UniProt places it at endosomes, the trans-Golgi network, vesicles and cilia (UniProt Q01968). In paraffin-section IHC, expect mainly cytoplasmic staining in HPA-listed cells, including glandular, respiratory epithelial and neuronal cells (HPA tissue IHC). HPA calls the tissue pattern Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in adrenal gland or breast glandular cells. | This fits the general cytoplasmic profile and the High staining reported for those cells (HPA tissue IHC). Record the cell type and intensity rather than treating every positive cell as equally informative; HPA reports low tissue specificity (HPA tissue IHC). |
| Cytoplasmic staining in bronchial respiratory epithelium, or in caudate or cerebral-cortex neurons. | These are additional High-staining cell populations (HPA tissue IHC). Vesicular or perinuclear detail may be compatible with UniProt endosome and trans-Golgi locations, but a routine chromogenic section cannot identify an organelle from appearance alone (UniProt Q01968; general IHC practice). |
| Predominantly nuclear staining with little cytoplasmic signal. | This is discordant with the HPA general cytoplasmic IHC profile and UniProt subcellular locations (HPA tissue IHC; UniProt Q01968). Review controls and staining conditions before assigning that compartment to OCRL; morphology alone cannot establish antibody specificity (general IHC practice). |
| Strong staining in an unexpected cell population, especially when expected cells are unstained. | Possible explanations include cross-reactivity or endogenous chromogenic activity (general IHC practice). HPA lists High staining in selected populations but also low tissue specificity and no negative cell examples in this payload, so staining elsewhere is not automatically false (HPA tissue IHC). |
| Uniform haze across tissue, or no staining in an HPA High-staining population. | Haze obscuring cell boundaries suggests background rather than a readable OCRL pattern (general IHC practice). An absent signal in a listed High-staining population warrants a run check, but HPA's Approved rating and medium RNA–staining consistency do not guarantee every specimen will stain (HPA tissue IHC). |
💡Expected OCRL appearanceCall a result concordant when discernible cytoplasmic signal occurs in the relevant HPA High-staining cells; diffuse haze or predominantly nuclear signal calls for control review (HPA tissue IHC; UniProt Q01968; general IHC practice).
| Tissue and cell selection | HPA reports High staining in adrenal, appendix and breast glandular cells; bronchial respiratory epithelial cells; bone-marrow hematopoietic cells; and selected brain cells (HPA tissue IHC). Match the assessment to the cell population actually present in the section (general IHC practice). |
| Subcellular resolution | UniProt lists endosomal, vesicular, trans-Golgi and ciliary locations (UniProt Q01968). HPA tissue IHC reports a general cytoplasmic profile, so do not require visible cilia or organelle-sized puncta to call a paraffin-section IHC result concordant (HPA tissue IHC; general IHC practice). |
| Antibody evidence | HPA012495 is Approved for IHC and Supported for ICC-IF; the overall tissue-IHC rating has medium staining–RNA consistency (HPA antibodies; HPA tissue IHC). These ratings support comparison with the reported pattern but do not establish specificity in every specimen (general IHC practice). |
| Isoforms and epitope | UniProt records two OCRL isoforms, A and B (UniProt Q01968). The supplied evidence gives no antibody epitope or isoform-specific staining claim, so an IHC difference cannot be assigned to one isoform from this record (UniProt Q01968; HPA antibodies). |
| Processing and fixation evidence | UniProt lists a single 1–901 chain, with no signal peptide, propeptide or glycosylation sites (UniProt Q01968). The supplied HPA and UniProt records do not report OCRL-specific fixation sensitivity; assess retrieval problems with ordinary run controls, without attributing them to OCRL (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in an HPA High-staining cell population. | The section, retrieval, primary-antibody step or detection run may have failed (general IHC practice); HPA does not establish an OCRL-specific fixation effect (HPA tissue IHC). | Confirm the target cells are present and inspect an appropriate positive-control section and run controls before changing one IHC condition at a time (general IHC practice). |
| Weak cytoplasmic staining is hard to score. | Low signal or excessive counterstain can reduce contrast (general IHC practice). HPA's High designation describes reported cells, not a guaranteed intensity in each specimen (HPA tissue IHC). | Compare equivalent cell populations with the positive control; check primary-antibody dilution, detection and counterstain against the validated workflow (general IHC practice). |
| Diffuse staining covers multiple compartments. | Nonspecific binding, excessive chromogen development or inadequate washing can obscure cellular boundaries (general IHC practice). | Review the no-primary control, blocking, washes and development time; score OCRL only where cell-associated staining remains distinguishable (general IHC practice; HPA tissue IHC). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). The supplied HPA profile has no negative cell examples and reports low tissue specificity (HPA tissue IHC). | Check the no-primary and detection controls, then compare cell morphology and distribution with HPA's reported populations before labeling the finding false-positive (general IHC practice; HPA tissue IHC). |
| Nuclear signal dominates the section. | The pattern conflicts with reported general cytoplasmic IHC staining and UniProt's listed locations (HPA tissue IHC; UniProt Q01968). Background or nonspecific recognition is possible (general IHC practice). | Inspect controls and re-evaluate the cytoplasmic compartment in HPA-listed cells; do not infer a new OCRL nuclear location from chromogenic staining alone (general IHC practice; HPA tissue IHC). |
| Can IF/ICC resolve an uncertain IHC compartment? | HPA ICC-IF supports primary-cilium localization; its microtubule, cytosol, centriolar-satellite and basal-body assignments are uncertain (HPA subcellular ICC-IF). These cell-image findings do not make cilia a required feature of tissue IHC (HPA tissue IHC). | Use the separate IF/ICC guide for that application; compare any IF localization claim with HPA's supported primary-cilium assignment and preserve the uncertainty of its other assignments (HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing OCRL staining
A02042-2 · IHCIHC analysis of OCRL using anti-OCRL antibody (A02042-2). OCRL was detected in a paraffin-embedded section of human colon adenocarcinoma tis
A02042-2 · IHCIHC analysis of OCRL using anti-OCRL antibody (A02042-2). OCRL was detected in a paraffin-embedded section of human endometrioid adenocarcin
A02042-2 · IFIF analysis of OCRL using anti-OCRL antibody (A02042-2). OCRL was detected in an immunocytochemical section of A549 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122126-OCRL/tissue/cerebral+cortex#imid_3205760
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122126-OCRL/tissue/colon#imid_3205694
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122126-OCRL/tissue/liver#imid_3205695
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000122126-OCRL/tissue/kidney#imid_3205688
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA012495; green = OCRL, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA012495; green = OCRL, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for OCRL IHC & IF
🧪Run breast first: glandular cells should stain strongly (HPA: High in breast glandular cells). HPA detects OCRL in all 45 scored tissues, so there is no designated negative tissue; use no-primary and isotype controls, and judge cells with little apparent staining against those controls without assuming breast stromal cells are OCRL-negative (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: OCRL 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 OCRL in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Microtubules (uncertain), Primary cilium (supported) (HPA subcellular).
Technical controls: Include a secondary-only slide, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where known, and an OCRL-knockout sample as a biological negative (selected-SKU caption: rabbit anti-OCRL antibody; standard IHC practice). Quench endogenous peroxidase and check background in the breast section before interpreting DAB staining (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). The reported IHC conditions use heat retrieval in EDTA at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C; retrieval dependence has not been established by a comparison in the supplied evidence (selected-SKU caption). ICC-IF images exist, but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC; assess breast-section background because HRP/DAB detection can be affected by endogenous peroxidase (HPA: ICC-IF image cell lines; selected-SKU caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for OCRL
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: OCRL is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced OCRL IHC Tips
Troubleshoot chromogenic OCRL staining in paraffin sections using the catalog antibody’s tissue result, reported localisation, and appropriate controls.
What retrieval should I try first when OCRL staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02042-2). The catalog antibody detected OCRL in a paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A02042-2). Compare retrieved and unretrieved serial sections, holding antibody concentration, incubation and DAB development constant so the effect of retrieval is interpretable (standard IHC practice). If staining remains weak, optimize heating time on matched sections and check tissue integrity, since the caption supplies the buffer and pH but no heating duration (datasheet A02042-2; standard IHC practice).
Could fixation be hiding OCRL in my paraffin sections?
Target specific fixation sensitivity is unknown: the catalog tissue caption identifies a paraffin section but does not state its fixative or fixation duration (datasheet A02042-2). Record each specimen’s fixative and fixation time, and compare similarly processed sections before attributing weak DAB signal to OCRL biology (standard IHC practice). Run the reported EDTA pH 8.0 retrieval and 2 μg/ml overnight primary incubation on a control section in the same staining run (datasheet A02042-2). If fixation histories differ, evaluate staining and tissue preservation together; neither the reported tissue expression nor OCRL topology establishes a fixation effect (standard IHC practice; UniProt Q01968 topology).
Where should convincing OCRL staining appear in tissue?
Expect a predominantly cytoplasmic IHC pattern, while recognizing that compartment detail can be difficult to resolve with DAB in tissue sections (HPA tissue IHC: general cytoplasmic expression; standard IHC practice). OCRL is reported at endosomes, the trans Golgi network, lysosomes, coated pits and phagosomes, with a supported primary cilium location in subcellular imaging (UniProt Q01968 subcellular location; HPA subcellular). Its sequence has no transmembrane segment, so a uniformly sharp plasma membrane outline alone needs additional validation before assignment to OCRL (UniProt Q01968 topology; standard IHC practice). Compare the pattern with glandular and hematopoietic cells reported as strongly stained, while assessing each cell type separately (HPA tissue IHC).
Can this stain distinguish OCRL isoforms or reveal an inaccessible epitope?
OCRL has 2 reported isoforms, A and B, but the supplied catalog caption does not map this antibody’s epitope or establish isoform selectivity (UniProt Q01968 isoforms; datasheet A02042-2). Its annotated PH, 5-phosphatase and Rho-GAP regions provide structural context, not evidence that a particular region is recognized in paraffin sections (UniProt Q01968 domains; datasheet A02042-2). Compare staining after the reported EDTA pH 8.0 retrieval on matched sections before interpreting a missing cell population as isoform specific (datasheet A02042-2; standard IHC practice). An isoform claim requires independent epitope documentation or isoform specific validation beyond this IHC result (standard IHC practice).
How should I check OCRL localisation by multiplex IF?
Use IF as a complementary localisation check and include a marker that identifies the cell population scored in the chromogenic section; HPA reports strong OCRL staining in glandular and hematopoietic cells (HPA tissue IHC; standard IF practice). Choose fluorophores and filter sets after inspecting unstained tissue autofluorescence, and include single stain controls to assess signal bleed through (standard IF practice). For an intracellular OCRL epitope, test permeabilisation appropriate to cytosolic access while preserving endosomal and ciliary structures; the antibody epitope is unspecified here (UniProt Q01968 subcellular location; datasheet A02042-2; standard IF practice). Treat IF staining and the reported pH 8.0 paraffin IHC retrieval as separate assay conditions (datasheet A02042-2; standard IF practice).
How can I separate OCRL staining from DAB background?
The reported tissue result used 10% goat serum blocking, a peroxidase conjugated secondary antibody and DAB detection (datasheet A02042-2). Add a primary omission control and a suitable endogenous peroxidase block, then compare deposits in tissue, vessels and section edges with the complete stain (standard IHC practice). Keep the reported 2 μg/ml primary concentration and overnight 4°C incubation as a reference while adjusting one staining variable at a time (datasheet A02042-2; standard IHC practice). A broad cytoplasmic pattern alone does not prove specificity because HPA reports general cytoplasmic expression with only medium consistency against RNA data (HPA tissue IHC).
How should I quantify OCRL across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, since HPA reports general cytoplasmic expression and strong staining in several distinct cell types (HPA tissue IHC; standard IHC practice). For cytoplasmic DAB, report the percentage of positive cells and an H-score based on intensity and positive fraction; use positive cell density per mm² when cell abundance is the question (standard IHC practice). Normalize counts to viable tissue area or the number of eligible cells in the same compartment, and exclude folds, edges and necrotic regions by a fixed rule (standard IHC practice). Keep retrieval, exposure to DAB, counterstain and scoring thresholds consistent across compared sections (standard IHC practice).
What would make an apparent OCRL positive result unconvincing?
A credible result should reproduce the catalog antibody’s paraffin section staining under EDTA pH 8.0 retrieval and fit the broadly cytoplasmic tissue pattern (datasheet A02042-2; HPA tissue IHC). OCRL also occupies intracellular trafficking compartments and the primary cilium, so isolated nuclear staining or a continuous surface outline warrants a specificity check (UniProt Q01968 subcellular location and topology; HPA subcellular; standard IHC practice). Evaluate the cell population against reported glandular and hematopoietic staining, without treating those examples as an exhaustive list of positive cells (HPA tissue IHC). Reject edge concentrated deposits, necrotic staining or signal persisting after primary omission as evidence of OCRL until technical background is resolved (standard IHC practice).
Boster reagents
Best OCRL / Inositol polyphosphate 5-phosphatase OCRL IHC Antibodies
The catalog antibody A02042-2 has IHC data from human paraffin sections and IF/ICC data from A549 cells (catalog image captions).
Real IHC data
Anti-OCRL Antibody ®
Cat # A02042-2
A02042-2 has IHC images from paraffin sections of human colon adenocarcinoma, endometrioid adenocarcinoma and liver cancer (A02042-2 IHC captions). It also has an IF/ICC image from A549 cells (A02042-2 IF caption).
Which to pick: Choose A02042-2 for human tissue IHC: its colon adenocarcinoma caption documents paraffin sections, EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A02042-2 IHC caption). For IF/ICC, A02042-2 has an A549 cell image, and its application list includes IF and ICC (A02042-2 IF caption; catalog applications). No cross-species or clonality-based choice is supported: the listed reactivity is Human and the clone field is unreported (A02042-2 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q01968 (OCRL_HUMAN, Inositol polyphosphate 5-phosphatase OCRL).
- Human Protein Atlas. OCRL tissue IHC expression (reliability: Approved).
- Human Protein Atlas. OCRL subcellular location (ICC-IF): Mainly localized to the microtubules and primary cilium. In addition localized to the cytosol, centriolar satellites and basal body..
- Human Protein Atlas. OCRL antibody validation summary (1 antibodies).
- Compensatory Role of Inositol 5-Phosphatase INPP5B to OCRL in Primary Cilia Formation in Oculocerebrorenal Syndrome of Lowe. PloS one 2013 — PMC3689662.
- Dysregulation of Inositol Polyphosphate 5-Phosphatase OCRL in Alzheimer's Disease: Implications for Autophagy Dysfunction. International journal of molecular sciences 2025 — PMC12192959.
- PubMed PMID:1321346 — UniProt-cited evidence.
- PubMed PMID:9048911 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for OCRL (Inositol polyphosphate 5-phosphatase OCRL). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02042-2). Validated IHC antibody: A02042-2 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 A02042-2); 2. Fixation: Image fixative and duration unreported (datasheet A02042-2); 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 A02042-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02042-2); 8. Primary antibody: Rabbit anti-OCRL, 2-5 μg/ml (datasheet A02042-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02042-2). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02042-2); Caveat: Staining and RNA levels show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms (A/B); epitope coverage unknown (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q01968, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the microtubules and primary cilium. In addition…, Datasheet A02042-2, PMC12192959. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/OCRL/immunohistochemistry-ihc-guide.