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HIP1R / Huntingtin-interacting protein 1-related protein · IHC design guide
Design Immunohistochemistry for HIP1R
Plan HIP1R staining in paraffin sections using the observed cytoplasmic and occasional membranous tissue pattern (HPA tissue IHC). Compare cell types and include controls when interpreting staining intensity.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HIP1R 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 with some membranous staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells: cytoplasmic, sometimes membranous staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05059-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended HIP1R IHC & IF Protocols
The catalog antibody protocol (datasheet A05059-2) is paired with published HIP1R IHC protocols for diffuse gliomas (PMC12063079) and lung cancer (PMC7291156).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05059-2) |
| Fixation | Image fixative and duration unreported (datasheet A05059-2); 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 A05059-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05059-2) |
| Primary antibody | Rabbit anti-HIP1R, 2-5 μg/ml (datasheet A05059-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05059-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05059-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HIP1R-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression combined with membranous expression in several tissues. No signal in the no-primary control. |
💡Decision noteFor the catalog antibody, start with heat-mediated EDTA pH 8.0 retrieval (datasheet A05059-2). Neither published excerpt specifies a retrieval method (PMC12063079; PMC7291156).
Section 2
What Is the Expected HIP1R Staining Pattern?
HIP1R is membrane-associated at endocytic compartments and the perinuclear region, with no transmembrane segment (UniProt O75146 localization and topology). In paraffin-section IHC, expect cytoplasmic staining with membranous staining in some tissues (HPA: tissue IHC profile). High staining is reported in several glandular cell populations, bronchial goblet cells, cerebellar Purkinje cells, and fallopian-tube ciliated cell bodies (HPA: High). HPA rates tissue IHC reliability Enhanced, while reporting medium agreement with RNA expression (HPA: reliability).
| Cytoplasmic staining, sometimes with a membrane-associated component, is concentrated in the expected cells. | This fits the tissue IHC profile (HPA: general cytoplasmic expression with membranous expression in several tissues). Examples of strongly stained cells include colon and duodenal glandular cells and cerebellar Purkinje cells (HPA: High). Assess the named cell population, since neighboring cell types need not share its staining level (HPA: cell-specific tissue IHC observations). |
| The dominant signal is nuclear, with little cytoplasmic or membranous staining. | A dominant nuclear pattern conflicts with the reported compartments (UniProt O75146 localization; HPA: tissue IHC profile). Treat it as suspect, then compare morphology, the no-primary control, and an expected-positive area before assigning HIP1R positivity (general IHC practice). |
| Strong staining appears in adipocytes or skeletal-muscle myocytes. | Those specific cell populations were not detected in the HPA tissue observations (HPA: adipocytes and skeletal-muscle myocytes, Not detected). Check the no-primary control for endogenous detection activity and reassess antibody specificity if the signal persists (general IHC practice). A negative entry for one cell population does not classify every cell in that tissue (HPA: cell-specific tissue IHC observations). |
| Brown color spreads across unrelated structures and obscures cell boundaries. | This prevents a reliable compartment or cell-type call (general IHC practice). Possible contributors include nonspecific antibody binding, endogenous detection activity, or excessive chromogen development (general IHC practice). Compare controls and tissue morphology before calling the diffuse color HIP1R staining (general IHC practice). |
| No staining is visible in an expected-positive cell population. | First confirm that the selected population is actually present: appendix glandular cells and bronchial goblet cells are reported High, for example (HPA: tissue IHC). An absent signal warrants checking the IHC-validated antibody, detection reagents, and run controls (general IHC practice). HPA's Enhanced rating supports the observed pattern but does not guarantee signal in every preparation (HPA: reliability). |
💡Expected HIP1R appearanceCall a positive when expected cells show clear cytoplasmic, sometimes membranous, staining—potentially High in listed populations (HPA: tissue IHC); dominant nuclear-only color or strong staining in listed Not detected cells is suspect (UniProt O75146 localization; HPA: tissue IHC).
| Compartment and topology | HIP1R is associated with clathrin-coated vesicle membranes and the perinuclear endomembrane system, without a transmembrane segment (UniProt O75146 localization and topology). Interpret a membranous component alongside cytoplasmic staining; do not require a sharp, continuous cell-surface rim to recognize the reported IHC pattern (HPA: tissue IHC profile). |
| Tissue and cell selection | HPA reports High staining in named glandular, goblet, Purkinje, and ciliated cell populations, but Not detected in specified adipocytes, myocytes, smooth-muscle cells, chondrocytes, and parathyroid glandular cells (HPA: tissue IHC). Select controls by cell population rather than tissue name alone (general IHC practice). |
| Evidence strength | HPA038135 and HPA038136 have Enhanced IHC validation, whereas CAB017187 is Approved (HPA: antibody validation). The tissue profile still has medium consistency with RNA expression (HPA: reliability). Use those ratings as context when weighing an unexpected result, rather than treating RNA or one stained section as decisive. |
| Isoforms and antibody coverage | UniProt lists 2 HIP1R isoforms (UniProt O75146 isoforms). Their existence alone does not establish which isoforms a given IHC antibody recognizes; resolve an isoform-specific question from that antibody's documented epitope or validation data before interpreting a discordant stain (general IHC practice). |
| IF/ICC Q: what localization is expected? | HPA reports mainly plasma membrane and cytosol, with additional vesicles, in its ICC-IF summary (HPA: subcellular ICC-IF). This supports a compartment comparison, while the paraffin-section chromogenic IHC call should follow the tissue IHC profile (HPA: tissue IHC). IF/ICC conditions belong to its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive cells are blank. | The population may be absent from the section, or an IHC staining step may have failed (general IHC practice). | Confirm the cells morphologically, then check a run control and the IHC-validated antibody's documented paraffin-section conditions (general IHC practice). Use a listed High population, such as colon glandular cells, when available (HPA: tissue IHC). |
| Only nuclei stain strongly. | The compartment conflicts with reported cytoplasmic and membrane-associated localization (UniProt O75146 localization; HPA: tissue IHC). | Inspect the counterstain and no-primary control; compare with the expected cytoplasmic pattern before scoring positive cells (general IHC practice; HPA: tissue IHC profile). |
| Listed Not detected cell populations stain strongly. | Nonspecific binding or endogenous detection activity is possible (general IHC practice); the result conflicts with the named HPA observations (HPA: tissue IHC). | Review cell identity and the no-primary control, then assess antibody specificity with the available validation information (general IHC practice; HPA: antibody validation). |
| Background masks a weak cellular pattern. | Nonspecific binding, endogenous detection activity, or overdeveloped chromogen can reduce interpretability (general IHC practice). | Use the relevant blocking and detection controls, review antibody dilution and development time, and score only clearly localized cellular signal (general IHC practice). |
| One tissue appears weaker than expected from a broad expression statement. | The records differ in granularity: UniProt lists tissue specificity, while HPA records staining levels for named cell types (UniProt O75146 tissue specificity; HPA: tissue IHC). | Compare the exact cell population with its HPA entry. For example, heart cardiomyocytes are Low and placental decidual cells are Low in the supplied HPA observations (HPA: tissue IHC). |
| Membranous staining seems inconsistent between IHC and IF images. | The summaries emphasize different observations: IHC is generally cytoplasmic with membranous staining in several tissues, while ICC-IF highlights plasma membrane, cytosol, and vesicles (HPA: tissue IHC; HPA: subcellular ICC-IF). | Interpret each image by its stated application and compare cell identity and compartment before calling disagreement (HPA: tissue IHC; HPA: subcellular ICC-IF; general IHC practice). |
Experiment evidence
IHC & IF images showing HIP1R staining
A05059-2 · IHCIHC analysis of HIP1R using anti-HIP1R antibody (A05059-2). HIP1R was detected in a paraffin-embedded section of human liver cancer tissue.
A05059-2 · IHCIHC analysis of HIP1R using anti-HIP1R antibody (A05059-2). HIP1R was detected in a paraffin-embedded section of human lung cancer tissue. H
A05059-2 · IFIF analysis of HIP1R using anti-HIP1R antibody (A05059-2). HIP1R was detected in an immunocytochemical section of MCF-7 cells. Enzyme antige
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130787-HIP1R/tissue/cerebral+cortex#imid_15421411
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000130787-HIP1R/tissue/salivary+gland#imid_15421351
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130787-HIP1R/tissue/colon#imid_15416585
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130787-HIP1R/tissue/liver#imid_15416570
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017187; green = HIP1R, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017187; green = HIP1R, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HIP1R IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, adjacent nonglandular cells should lack specific chromogen staining, but their HIP1R-negative status is unverified.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show HIP1R in A-431, U-251MG, U2OS, MCF-7, with annotated localisation: Plasma membrane (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and matched rabbit IgG isotype controls; match the primary’s clonality, which the caption does not specify (selected IHC caption: rabbit primary). If available, use HIP1R-knockout material or a validated peptide block as a biological specificity control; quench endogenous peroxidase and check endogenous biotin because the caption uses biotin-based DAB detection (selected IHC caption).
⚠️Feasibility: The selected paraffin-section caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected IHC caption). A target-specific fixation window and fixation effect are unreported, and the fixative for SKU A05059-2 is unreported (selected IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; in colon, inspect luminal debris for misleading DAB deposits (standard IHC practice).
HPA tissue IHC evidence for HIP1R
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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Goblet cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Chondrocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HIP1R IHC Tips
Troubleshoot HIP1R chromogenic IHC by checking retrieval, cytoplasmic and membranous staining, cell identity, and matched controls.
What retrieval should I try first if HIP1R staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A05059-2). The catalog antibody stained a paraffin-embedded human liver cancer section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A05059-2). If staining is weak, check heating consistency, section adhesion, and whether the primary incubation matches that demonstrated workflow before changing retrieval conditions (standard IHC practice). Compare any retrieval adjustment on adjacent sections with the same detection and counterstain, so a stronger signal can be judged against tissue damage and background (standard IHC practice).
Could fixation explain weak or uneven HIP1R staining?
The selected paraffin-section caption does not state a fixative, so HIP1R-specific sensitivity to fixation is unknown (datasheet A05059-2). Record each specimen’s fixative and processing history, then compare similarly processed sections before attributing a staining difference to HIP1R biology (standard IHC practice). If signal varies across a section, inspect morphology and staining at folds and edges, and repeat on an intact section using EDTA retrieval at pH 8.0 (datasheet A05059-2; standard IHC practice). Keep the demonstrated 2 μg/ml overnight primary incubation constant while assessing processing differences, and include a matched control section in the run (datasheet A05059-2; standard IHC practice).
Which staining compartments are plausible for HIP1R in tissue?
Expect chiefly cytoplasmic staining, potentially with membranous accentuation, in chromogenic tissue IHC (HPA tissue IHC: general cytoplasmic and membranous expression). HIP1R is associated with endocytic compartments, clathrin-coated vesicles, and the perinuclear region, and it has no transmembrane segment (UniProt O75146 localisation and topology). A crisp surface pattern alone should therefore be checked alongside cytoplasmic staining and tissue morphology rather than assigned to an integral membrane pool (UniProt O75146 topology; standard IHC practice). Compare candidate positive cells with nearby internal controls: glandular cells in colon are reported as high, whereas adipocytes are reported as not detected (HPA tissue IHC).
Can a negative stain exclude both HIP1R isoforms?
No: HIP1R has 2 reported isoforms, while the supplied IHC caption does not identify the catalog antibody’s epitope or isoform coverage (UniProt O75146 isoforms; datasheet A05059-2). The protein contains an ENTH domain at residues 23–151 and an I/LWEQ domain at 771–1012, but domain positions alone cannot establish antibody recognition (UniProt O75146 domains; standard IHC interpretation). Check the antibody’s documented immunogen or epitope against each isoform before making an isoform-specific claim (standard IHC practice). If that information is unavailable, report staining as antibody-detected HIP1R and interpret a negative section within the limits of retrieval and controls (standard IHC practice).
How can IF help resolve ambiguous HIP1R IHC staining?
Use IF as a separate follow-up when chromogenic IHC cannot resolve which cells or compartments carry signal (standard microscopy practice). Multiplex HIP1R with a validated marker for the cell type being examined, and compare the pattern with HIP1R’s reported cytosolic, plasma-membrane, and vesicular locations (HPA subcellular; standard IF practice). Choose fluorophores after inspecting the tissue’s autofluorescence and include single-label controls to assess bleed-through (standard IF practice). If the antibody epitope faces the cytosol, optimise permeabilisation for access to that epitope; HIP1R has no transmembrane segment, but the supplied evidence does not establish this antibody’s epitope orientation (UniProt O75146 topology; standard IF practice).
How do I reduce diffuse brown background without losing HIP1R signal?
First distinguish widespread background from the expected cytoplasmic staining with possible membranous accentuation (HPA tissue IHC). The demonstrated tissue workflow used 10% goat serum, a biotinylated secondary antibody, an avidin-biotin detection complex, and DAB (datasheet A05059-2). Include a no-primary control, assess the need for a peroxidase block, and check whether the detection system contributes signal independently of primary antibody (standard chromogenic IHC practice). If background persists, compare adjacent sections while changing one factor at a time, such as blocking or primary-antibody concentration, using the demonstrated 2 μg/ml condition as a reference (datasheet A05059-2; standard IHC practice).
How should I score HIP1R staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HIP1R tissue staining is generally cytoplasmic with membranous expression in several tissues (HPA tissue IHC). For comparable chromogenic sections, report the percentage of positive target cells and an H-score combining intensity with that percentage; keep scoring thresholds fixed across the batch (standard IHC quantification practice). If spatial abundance is the question, report positive cells per mm² of viable tissue and identify the counted compartment (standard pathology quantification practice). Normalise to eligible cells or viable tissue area, exclude folds and necrosis, and review section quality and batch controls before interpreting between-specimen differences (standard IHC practice).
What makes a HIP1R-positive IHC pattern convincing?
Look for staining in intact cells with a plausible cytoplasmic or membranous pattern, supported by a clean no-primary control (HPA tissue IHC; standard IHC practice). Cell identity matters: HPA reports high staining in appendix glandular cells and cerebellar Purkinje cells, while skeletal-muscle myocytes are reported as not detected (HPA tissue IHC). Treat signal confined to section edges, folds, or necrotic regions cautiously, and check for staining caused by endogenous enzyme activity in the chromogenic workflow (standard IHC practice). HPA rates its tissue staining reliability as enhanced but reports medium consistency with RNA expression, so a single brown signal should be interpreted alongside morphology and independent controls (HPA tissue IHC).
Boster reagents
Best HIP1R / Huntingtin-interacting protein 1-related protein IHC Antibodies
A05059-2 has IHC data from human paraffin sections (IHC image captions) and IF/ICC data from MCF-7 cells (IF image caption); catalog reactivity includes human, mouse, rat and monkey (catalog reactivity).
Real IHC data
Anti-HIP1R Antibody ®
Cat # A05059-2
A05059-2 has IHC images from human paraffin sections of liver cancer, lung cancer, rectal cancer and renal clear cell carcinoma (IHC image captions). A05059-2 also has IF/ICC data in MCF-7 cells (IF image caption) and lists human, mouse, rat and monkey reactivity (catalog reactivity).
Which to pick: Choose A05059-2 for human paraffin-section IHC because its own IHC captions show that preparation; the fixative is unreported (IHC image captions). Choose A05059-2 for IF/ICC based on its MCF-7 image and listed IF/ICC applications (IF image caption; catalog applications). For other species, A05059-2 lists mouse, rat and monkey reactivity, but the supplied IHC and IF images show human samples only; clonality is unreported.
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75146 (HIP1R_HUMAN, Huntingtin-interacting protein 1-related protein).
- Human Protein Atlas. HIP1R tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HIP1R subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to vesicles..
- Human Protein Atlas. HIP1R antibody validation summary (3 antibodies).
- Evaluating the efficacy of Hip1R, Vimentin, and H3K27me3 as surrogate markers for 1p/19q co-deletion in oligodendrogliomas. Neuro-oncology advances 2025 — PMC12063079.
- HIP1R Expression and Its Association with PD-1 Pathway Blockade Response in Refractory Advanced NonSmall Cell Lung Cancer: A Gene Set Enrichment Analysis. Journal of clinical medicine 2020 — PMC7291156.
- Therapeutic implications of cancer-associated fibroblast heterogeneity: insights from single-cell and multi-omics analysis. Frontiers in immunology 2025 — PMC12206818.
- Flurbiprofen inhibits cell proliferation in thyroid cancer through interrupting HIP1R-induced endocytosis of PTEN. European journal of medical research 2022 — PMC8867849.
- PubMed PMID:11063258 — UniProt-cited evidence.
- PubMed PMID:9734811 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HIP1R (Huntingtin-interacting protein 1-related protein). Expected localisation Cytoplasmic staining with some membranous staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05059-2). Validated IHC antibody: A05059-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A05059-2); 2. Fixation: Image fixative and duration unreported (datasheet A05059-2); 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 A05059-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05059-2); 8. Primary antibody: Rabbit anti-HIP1R, 2-5 μg/ml (datasheet A05059-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05059-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt O75146, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A05059-2, PMC12063079, PMC7291156. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HIP1R/immunohistochemistry-ihc-guide.