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HYOU1 / Hypoxia up-regulated protein 1 · IHC design guide
Design Immunohistochemistry for HYOU1
Plan HYOU1 staining in paraffin sections using its observed cytoplasmic tissue pattern and expected ER localisation (HPA tissue IHC; UniProt). Use pancreatic exocrine cells as a high-staining reference, while accounting for the reported low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HYOU1 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 | Mainly cytoplasmic in tissue (HPA tissue IHC); ER lumen expected (UniProt) | |
| Staining pattern | Cytoplasmic staining in pancreatic exocrine cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04934-2) | |
| Positive control | Pancreas+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Up-regulated by hypoxia (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage for each (UniProt) |
Section 1
Recommended HYOU1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A04934-2) is accompanied by one published nasopharyngeal carcinoma IHC protocol (PMC4749358).
| Sample | Paraffin-embedded human gall bladder adenosquamous carcinoma tissue; fixative not specified (datasheet A04934-2) |
| Fixation | Image fixative and duration unreported (datasheet A04934-2); 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 A04934-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04934-2) |
| Primary antibody | Rabbit anti-HYOU1, 1-2 μg/ml (datasheet A04934-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04934-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04934-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HYOU1-positive staining in exocrine glandular cells of pancreas (HPA tissue IHC: High). HPA tissue profile: Mainly cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A04934-2); the published protocol does not state retrieval conditions (PMC4749358).
Section 2
What Is the Expected HYOU1 Staining Pattern?
HYOU1 is a soluble endoplasmic reticulum lumen protein with no transmembrane segment (UniProt Q9Y4L1 topology). In paraffin section IHC, expect mainly cytoplasmic staining, especially in pancreatic exocrine glandular cells, where HPA reports high staining (HPA tissue IHC). Interpret that expectation cautiously: HPA rates the tissue staining Approved but reports low consistency between antibody staining and RNA expression data (HPA tissue IHC).
| Pancreatic exocrine glandular cells show strong cytoplasmic chromogen, while nearby cells vary in intensity. | This fits the clearest listed positive tissue and the mainly cytoplasmic tissue profile (HPA tissue IHC: high in pancreatic exocrine glandular cells). Variation between cells need not invalidate the section; evaluate the expected cell population and compartment before assigning a score. |
| The dominant signal is nuclear, membranous, or restricted to an unexpected compartment. | That pattern conflicts with an ER lumen protein seen mainly as cytoplasmic staining in tissue IHC (UniProt Q9Y4L1 topology; HPA tissue IHC). Treat it as a possible staining artefact and check the antibody and detection controls before calling it HYOU1. |
| Prominent staining appears in cells listed as undetected, such as adipocytes in adipose tissue. | This conflicts with that specific HPA observation and may reflect cross-reactivity or endogenous detection activity (HPA tissue IHC: adipocytes not detected; general IHC practice). It does not establish that every adipocyte in every specimen lacks HYOU1; compare a control section and review cellular morphology. |
| Chromogen covers stroma, empty spaces, and many cell types with little cellular contrast. | A diffuse deposit cannot be assigned confidently to the reported cytoplasmic pattern (HPA tissue IHC). In chromogenic IHC, inadequate blocking, nonspecific antibody binding, or detection background can produce this appearance (general IHC practice); judge the section against a negative control. |
| Pancreatic exocrine glandular cells show no convincing signal. | The expected high staining is absent (HPA tissue IHC: pancreatic exocrine glandular cells high). First check tissue preservation and whether a separate positive control stained; then review retrieval, antibody application, and detection steps (general IHC practice). A single blank section does not establish biological absence. |
💡Expected HYOU1 appearanceCall a convincing positive when pancreatic exocrine glandular cells show strong, mainly cytoplasmic staining (HPA tissue IHC: high; mainly cytoplasmic); dominant nuclear, membranous, or widespread cell-free signal is suspect against the ER lumen assignment (UniProt Q9Y4L1 topology; general IHC practice).
| Compartment and topology | HYOU1 is assigned to the ER lumen and has no transmembrane segment (UniProt Q9Y4L1 topology). In tissue IHC, HPA describes mainly cytoplasmic expression; interpret chromogen as a compartment-level pattern, not proof that individual ER structures are resolved (HPA tissue IHC; general IHC practice). |
| Tissue and cell context | HPA reports high staining in pancreatic exocrine glandular cells and medium staining in several neuronal and glandular populations (HPA tissue IHC). Select and score the named cell population: a tissue label alone does not predict equal staining in every cell. |
| Strength of the tissue evidence | The listed antibody HPA049296 has IHC status Approved; HPA also notes low consistency between antibody staining and RNA data (HPA antibody validation; HPA tissue IHC). Use the observed staining pattern as a working expectation and require appropriate controls for a new specimen. |
| IF/ICC expectation? | An ER lumen location suggests an intracellular signal if HYOU1 is specifically detected (UniProt Q9Y4L1 topology). HPA provides no main ICC-IF location or cell-line images in this payload, and lists no ICC status for HPA049296 (HPA subcellular; HPA antibody validation). The tissue IHC pattern alone cannot validate an IF/ICC result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Pancreatic exocrine glands are blank. | The expected high-staining population is absent from the readout (HPA tissue IHC); a failed staining run is one possible cause (general IHC practice). | Confirm the exocrine cells are present, inspect a separate positive control, then review antigen retrieval, antibody application, and detection in the run (general IHC practice). |
| Only nuclei or cell borders stain strongly. | The dominant compartment disagrees with ER lumen localisation and the mainly cytoplasmic tissue profile (UniProt Q9Y4L1 topology; HPA tissue IHC). | Compare control sections and reassess staining localisation under higher magnification before scoring HYOU1 (general IHC practice). |
| Adipocytes or other HPA-undetected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: adipocytes not detected; general IHC practice). | Check a negative control and the detection system; record the exact cell population rather than treating the whole tissue as positive (general IHC practice). |
| Diffuse colour obscures cell boundaries. | Nonspecific binding or detection background can obscure a cellular pattern (general IHC practice). | Review blocking, washing, antibody concentration, and detection timing against a negative control (general IHC practice). |
| A medium-staining tissue appears weaker than pancreas. | Different intensity is consistent with the supplied HPA levels, such as medium in cerebral cortical neurons versus high in pancreatic exocrine cells (HPA tissue IHC). | Score the specified cell types separately and compare sections processed in the same run before interpreting the difference (general IHC practice). |
| An IF/ICC image seems to show HYOU1 at the plasma membrane. | That location conflicts with the ER lumen assignment; this payload contains no HPA ICC-IF images or validated ICC location to resolve it (UniProt Q9Y4L1 topology; HPA subcellular). | Treat the IF/ICC localisation as unconfirmed and evaluate it with appropriate imaging and antibody controls in its separate guide (general IF practice). |
Experiment evidence
IHC & IF images showing HYOU1 staining
A04934-2 · IHCIHC analysis of ORP150/HYOU1 using anti-ORP150/HYOU1 antibody (A04934-2). ORP150/HYOU1 was detected in a paraffin-embedded section of human
A04934-2 · IHCIHC analysis of ORP150/HYOU1 using anti-ORP150/HYOU1 antibody (A04934-2). ORP150/HYOU1 was detected in a paraffin-embedded section of human
A04934-2 · IFIF analysis of ORP150/HYOU1 using anti-ORP150/HYOU1 antibody (A04934-2). ORP150/HYOU1 was detected in an immunocytochemical section of CACO-
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149428-HYOU1/tissue/cerebral+cortex#imid_13407123
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149428-HYOU1/tissue/colon#imid_13407057
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149428-HYOU1/tissue/liver#imid_13407058
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000149428-HYOU1/tissue/pancreas#imid_13407001
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HYOU1 IHC & IF
🧪Run pancreas first and look for staining in exocrine glandular cells (HPA: High in pancreatic exocrine glandular cells). Use adipose tissue as the negative comparison, with adipocytes expected to show no detectable staining (HPA: Not detected in adipocytes); on the pancreas slide, any cells used as internal negatives should show only background chromogen, without assigning an unreported pancreatic cell type as negative.
Positive control tissue: Pancreas (Exocrine glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for HYOU1; derive a cell-line control from the positive tissue's cell type (Exocrine glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a host- and format-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody); confirm specificity with HYOU1 knockout material or a validated peptide-block control. Quench endogenous peroxidase and check for endogenous biotin before using the caption’s biotinylated secondary, streptavidin–biotin complex and DAB detection (selected-SKU caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The reported IHC procedure uses heat retrieval in EDTA at pH 8.0, followed by 2 μg/ml primary antibody overnight at 4°C; this documents a workable retrieval condition, not a requirement for all specimens (selected-SKU caption). Frozen sections and IF/ICC are not established as easier by the supplied application evidence; for pancreatic chromogenic IHC, check biotin-related background with the reported SABC detection (selected-SKU caption: SABC/DAB).
HPA tissue IHC evidence for HYOU1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HYOU1 IHC Tips
These questions address chromogenic HYOU1 staining in paraffin sections, with one entry on IF/ICC and attention to its ER localisation (UniProt Q9Y4L1).
What retrieval conditions should I start with for HYOU1 staining in paraffin sections?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections stained with A04934-2 (datasheet A04934-2). Run a known positive section alongside the test material, and keep heating and cooling conditions consistent across slides so differences can be attributed to the sample (standard IHC practice). If staining remains weak, compare a citrate buffer at pH 6.0 as a fallback on matched sections, while checking whether tissue morphology or nonspecific staining worsens (standard IHC practice). Judge retrieval by cytoplasmic staining in the intended cells; HYOU1 is an ER-lumen protein, so intense nuclear staining is not the expected pattern (UniProt Q9Y4L1 topology).
How should I assess whether fixation is affecting HYOU1 staining?
Target-specific fixation sensitivity is unknown: the selected A04934-2 image identifies a paraffin section but does not report its fixative (caption A04934-2). Record the fixative, time in fixative, tissue thickness and processing history for each specimen, then compare matched sections under the same pH 8.0 EDTA retrieval conditions (standard IHC practice; datasheet A04934-2). Include a previously successful control section in the same staining run to help distinguish a specimen issue from a run-wide failure (standard IHC practice). Do not infer fixation tolerance from HPA staining patterns or HYOU1's ER topology and glycosylation; neither establishes how this antibody responds to fixation (HPA tissue IHC; UniProt Q9Y4L1).
Which staining pattern is plausible for HYOU1 in tissue sections?
Expect a cytoplasmic pattern compatible with the endoplasmic reticulum: HYOU1 resides in the ER lumen and has no annotated transmembrane segment (UniProt Q9Y4L1 topology). HPA describes mainly cytoplasmic staining across several tissue types, including high staining in pancreatic exocrine glandular cells (HPA tissue IHC). At routine chromogenic IHC resolution, judge the distribution within identified cells rather than claiming that diffuse brown signal resolves individual ER membranes (standard IHC practice). Compare glandular cells with adjacent tissue and a no-primary control; dominant nuclear, extracellular or uniformly surface-bound signal needs further specificity checks before being assigned to HYOU1 (UniProt Q9Y4L1 topology; standard IHC practice).
Could isoforms or epitope accessibility explain uneven HYOU1 staining?
HYOU1 has 2 annotated isoforms and a signal peptide at residues 1–32; the mature chain spans residues 33–999 (UniProt Q9Y4L1). The supplied A04934-2 caption does not identify its binding epitope, so staining cannot establish which isoform or processed region the antibody recognises (caption A04934-2; UniProt Q9Y4L1). HYOU1 also has 9 annotated glycosylation sites, but their effect on this antibody's tissue staining is untested here (UniProt Q9Y4L1). If a specimen gives unexpected patchiness, compare matched sections under the documented EDTA pH 8.0 retrieval and seek independent epitope or antibody validation before attributing the pattern to an isoform (datasheet A04934-2; standard IHC practice).
How can I check HYOU1 localisation by multiplex IF/ICC?
For an IF/ICC experiment, pair HYOU1 with a validated marker of the expected cell type, such as pancreatic exocrine glandular cells, and assess signal within marker-positive cells (HPA tissue IHC; standard IF practice). Choose a longer-wavelength fluorophore when tissue autofluorescence affects shorter channels, and include single-stain and no-primary controls to assess bleed-through and background (standard IF practice). Because HYOU1 is ER-luminal, optimise permeabilisation for antibody access across the plasma and ER membranes while monitoring cell morphology (UniProt Q9Y4L1 topology; standard IF practice). The supplied HPA subcellular record lists no ICC/IF images, so confirm any fluorescent pattern independently before treating it as validated localisation for this antibody (HPA subcellular).
What should I check when the DAB stain looks widespread or granular?
Start with a no-primary control and inspect where brown signal occurs; endogenous peroxidase, nonspecific secondary binding and precipitated DAB can complicate chromogenic IHC (standard IHC practice). Apply a peroxidase block and check reagent cleanliness and chromogen development under the same conditions used for the test slides (standard IHC practice). The A04934-2 example used 10% goat serum, biotinylated goat anti-rabbit secondary antibody and an avidin–biotin detection complex, so evaluate secondary-only and endogenous-biotin background when following that workflow (caption A04934-2; standard IHC practice). Titrate the primary around the reported 2 μg/ml only against positive and negative controls, preserving interpretable cell morphology (caption A04934-2; standard IHC practice).
How should I score HYOU1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then record the percentage of viable cells at each cytoplasmic intensity from 0–3 (standard IHC practice; UniProt Q9Y4L1 topology). Calculate an H-score from 0–300, or report percentage-positive cells with a stated threshold, using the same rule for every specimen (standard IHC practice). For a spatial analysis, report positive-cell density per mm² of viable tissue and exclude necrosis, folds and section edges from the measured area (standard IHC practice). Normalise comparisons to the relevant cell population or viable area, and use a common control section to monitor staining-batch variation; HPA reports low consistency between antibody staining and RNA expression (standard IHC practice; HPA tissue IHC).
How do I distinguish credible HYOU1 staining from artefact?
A credible result is cell-associated cytoplasmic staining compatible with HYOU1's ER-lumen localisation, interpreted in the identified cell population (UniProt Q9Y4L1 topology; standard IHC practice). Pancreatic exocrine glandular cells provide a reported high-staining reference, whereas adipocytes in adipose tissue are reported as not detected; neither reference alone proves antibody specificity (HPA tissue IHC; standard IHC practice). Investigate predominantly nuclear or extracellular staining, staining confined to section edges or necrosis, and signal in the no-primary control before calling a sample positive (UniProt Q9Y4L1 topology; standard IHC practice). Check peroxidase and, for the documented avidin–biotin workflow, endogenous-biotin background; HPA's approved tissue profile also carries a warning of low staining-to-RNA consistency (caption A04934-2; HPA tissue IHC; standard IHC practice).
Boster reagents
Best HYOU1 / Hypoxia up-regulated protein 1 IHC Antibodies
A04934-2 has human paraffin-section IHC images and a CACO-2 IF/ICC image (catalog image captions); its listed reactivity also includes Mouse and Rat (catalog: reactivity).
Real IHC data
Anti-ORP150/HYOU1 Antibody ®
Cat # A04934-2
A04934-2 is the SKU with a rendered IHC card, showing human gall bladder adenosquamous carcinoma in a paraffin section (catalog IHC image caption); its IF/ICC image shows CACO-2 cells (catalog IF image caption). M04934-2 lists IHC and Human, Mouse, Rat reactivity, but has no IHC or IF figure in the payload (catalog: applications/reactivity/image alts).
Which to pick: For tissue IHC, choose A04934-2 when a paraffin-section example is useful (catalog IHC image caption); the caption does not report the fixative (catalog IHC image caption). For IF/ICC, choose A04934-2 because it lists both applications and has a CACO-2 IF image (catalog: applications/IF image caption). Both SKUs list Human, Mouse, Rat reactivity; A04934-2 has pictured human IHC and IF data, while M04934-2 is monoclonal clone 19H99 with IHC listed but no accompanying figure (catalog: reactivity/clone/image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y4L1 (HYOU1_HUMAN, Hypoxia up-regulated protein 1).
- Human Protein Atlas. HYOU1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. HYOU1 subcellular location (ICC-IF): Highest expression in EFO-21: 476.0 nTPM.
- Human Protein Atlas. HYOU1 antibody validation summary (1 antibodies).
- Proteomic profiling of lung adenocarcinoma indicates heightened DNA repair, antioxidant mechanisms and identifies LASP1 as a potential negative predictor of survival. Clinical proteomics 2016 — PMC5084393.
- HYOU1, Regulated by LPLUNC1, Is Up-Regulated in Nasopharyngeal Carcinoma and Associated with Poor Prognosis. Journal of Cancer 2016 — PMC4749358.
- Unveiling FLNC variants: iPSC-derived myogenic cells as a model to study disease mechanisms. Skeletal muscle 2026 — PMC13063712.
- Acidic microenvironment plays a key role in human melanoma progression through a sustained exosome mediated transfer of clinically relevant metastatic molecules. Journal of experimental & clinical cancer research : CR 2018 — PMC6173926.
- PubMed PMID:9020069 — UniProt-cited evidence.
- PubMed PMID:10965054 — UniProt-cited evidence.
- PubMed PMID:17131193 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HYOU1 (Hypoxia up-regulated protein 1). Expected localisation Mainly cytoplasmic in tissue (HPA tissue IHC); ER lumen expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04934-2). Validated IHC antibody: A04934-2 at 1-2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Pancreas (Exocrine glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human gall bladder adenosquamous carcinoma tissue; fixative not specified…; 2. Fixation: Image fixative and duration unreported (datasheet A04934-2); 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 A04934-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04934-2); 8. Primary antibody: Rabbit anti-HYOU1, 1-2 μg/ml (datasheet A04934-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04934-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage for each (UniProt); Regulation: Up-regulated by hypoxia (UniProt). Evidence: UniProt Q9Y4L1, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in EFO-21: 476.0 nTPM, Datasheet A04934-2, PMC4749358. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HYOU1/immunohistochemistry-ihc-guide.