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P2RY6 / P2Y purinoceptor 6 · IHC design guide
Design Immunohistochemistry for P2RY6
Plan chromogenic IHC-P with the catalog antibody at 1:50 (datasheet M10087-1). Compare high staining in tonsil squamous epithelial cells with adipocytes where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
P2RY6 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 | Cell membrane expected (UniProt); tissue compartment unassigned (HPA tissue IHC) | |
| Staining pattern | Immune cell subsets in spleen, intestine and placental stroma (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M10087-1) | |
| Positive control | Tonsil+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 in unannotated cells complicates assignment (HPA tissue IHC) | |
| Regulation | Lymphoid tissue–enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | No isoforms or processing annotated; epitope side matters (UniProt) |
Section 1
Recommended P2RY6 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published P2RY6 chromogenic IHC workflows (datasheet; PMC10601148; PMC6844642; PMC13473056).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M10087-1) |
| Fixation | Image fixative and duration unreported (datasheet M10087-1); 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 M10087-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M10087-1) |
| Primary antibody | Rabbit monoclonal (clone 19P56) anti-P2RY6, 1:50 (datasheet M10087-1) |
| Primary incubation | Overnight at 4 °C (datasheet M10087-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M10087-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | P2RY6-positive staining in squamous epithelial cells of tonsil (HPA tissue IHC: High). HPA tissue profile: Expressed in a subset of immune cells in spleen, intestine and placental stroma. Also expressed in immune celler in other tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: M10087-1); citrate retrieval was reported with a different antibody (PMC13473056).
Section 2
What Is the Expected P2RY6 Staining Pattern?
P2RY6 is a seven-transmembrane receptor annotated at the cell membrane (UniProt Q15077 topology; UniProt Q15077 subcellular location). In IHC, expect staining in selected cells rather than every cell in a tissue: HPA reports positive immune-cell populations, along with several named epithelial and other cell populations (HPA tissue IHC). Treat these as reference patterns with limited certainty: HPA rates the tissue IHC evidence Enhanced but reports medium consistency with RNA expression (HPA tissue IHC).
| Selected cells show membrane-associated chromogenic staining; splenic red-pulp cells or tonsillar squamous epithelial cells are conspicuous. | This fits the predicted compartment (UniProt Q15077 cell membrane) and reported staining: medium in splenic red-pulp cells and high in tonsillar squamous epithelial cells (HPA tissue IHC). Evaluate the named cell population, since staining elsewhere in the same section does not establish the expected pattern (HPA tissue IHC). |
| The dominant signal is nuclear or uniformly fills cytoplasm, with no convincing membrane-associated component. | That distribution conflicts with the cell-membrane annotation (UniProt Q15077 subcellular location). Treat it as suspect staining and compare with a positive tissue and detection controls (general IHC practice); the supplied HPA tissue data do not establish a nuclear or diffuse cytoplasmic P2RY6 pattern (HPA tissue IHC). |
| Strong staining appears in cells HPA lists as not detected, such as adipocytes in adipose tissue or respiratory epithelial cells in bronchus. | This conflicts with those cell-specific reference observations (HPA tissue IHC). Check cell identification and controls for nonspecific primary binding or endogenous chromogen-generating activity (general IHC practice). A negative HPA observation is a comparator, not proof that every specimen of that tissue must be negative (HPA tissue IHC). |
| Chromogen spreads across tissue structures, including expected negative cell populations, without a cell-associated pattern. | Diffuse deposition is hard to reconcile with the selected-cell tissue profile (HPA tissue IHC) or predicted membrane location (UniProt Q15077 subcellular location). Assess background with an appropriate no-primary control, then review blocking, washing and detection chemistry (general IHC practice). |
| A known-positive cell population has no convincing signal, for example tonsillar squamous epithelial cells or splenic red-pulp cells. | That result disagrees with the reported high or medium HPA observations, respectively (HPA tissue IHC). First verify that the relevant cells are present and identifiable; then inspect the IHC run and its controls (general IHC practice). One negative section does not overturn an Enhanced but medium-consistency reference pattern (HPA tissue IHC). |
💡Expected P2RY6 appearanceCall a result positive when selected cells show credible membrane-associated staining, especially high staining in tonsillar squamous epithelial cells or medium staining in splenic red-pulp cells (UniProt Q15077 cell membrane; HPA tissue IHC); nuclear-only or broadly diffuse staining is suspect (UniProt Q15077 subcellular location; general IHC practice).
| Predicted compartment and epitope side | P2RY6 has seven transmembrane segments with extracellular and cytoplasmic regions (UniProt Q15077 topology). Its annotation supports a membrane expectation, but the supplied record gives no antibody epitope; do not infer which face the IHC-validated antibody recognizes or prescribe permeabilisation from topology alone (UniProt Q15077 topology; HPA antibodies). |
| Cell population within the section | HPA reports medium staining in appendix germinal-center cells, bone-marrow hematopoietic cells, lymph-node non-germinal-center cells and splenic red-pulp cells (HPA tissue IHC). Score the reported cell population, since a whole-section impression can miss a localized positive population (general IHC practice). |
| Reference strength | The listed antibody HPA074751 has Enhanced IHC validation, while HPA describes medium consistency between antibody staining and RNA expression (HPA antibodies; HPA tissue IHC). This supports using the reported pattern as a guide while investigating discordant cells rather than treating every listed intensity as a guaranteed outcome (HPA tissue IHC). |
| Lower-signal and negative comparators | HPA reports low staining in small-intestine goblet cells and no detection in several specified cell populations, including adrenal glandular cells and cerebellar granular-layer cells (HPA tissue IHC). These comparisons concern named cells, not blanket negativity throughout each tissue (HPA tissue IHC). |
| Sequence annotation limits | UniProt annotates one glycosylation site at residue 5 and no isoforms, signal peptide or propeptide in the supplied record (UniProt Q15077 processing and glycosylation). Those annotations do not identify the antibody epitope or establish a P2RY6-specific retrieval, fixation-sensitivity or shedding effect (UniProt Q15077 annotations). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells are absent or cannot be identified in the section. | The sampled area may lack the cell population needed to compare with the HPA reference (HPA tissue IHC). | Choose a section containing a reported positive population, such as tonsillar squamous epithelial cells or splenic red-pulp cells, and score those cells specifically (HPA tissue IHC; general IHC practice). |
| Reported positive cells are present, but the chromogenic IHC run shows no signal. | A failed or weak staining run is possible; the HPA reference alone cannot identify a target-specific fixation or retrieval cause (HPA tissue IHC). | Check section integrity, primary-antibody application, detection reagents and a run-level positive control; adjust general IHC conditions only against appropriate controls (general IHC practice). |
| Brown signal appears in unexpected negative-reference cells. | Nonspecific binding or endogenous detection activity can mimic a positive chromogenic result (general IHC practice); HPA lists some of these cell types as not detected (HPA tissue IHC). | Confirm cell identity, compare a no-primary control, and review blocking and detection steps before assigning P2RY6 positivity (general IHC practice). |
| Signal is widespread and obscures cell boundaries. | High background or deposited chromogen can make the membrane pattern unreadable (general IHC practice; UniProt Q15077 cell membrane). | Compare no-primary and positive controls; review primary-antibody concentration, washing and detection time as general IHC variables (general IHC practice). No target-specific dilution is supplied here. |
| A cell stains, but the signal is predominantly nuclear or diffuse cytoplasmic. | The compartment differs from the annotated cell-membrane location (UniProt Q15077 subcellular location). | Score it as inconclusive for the expected pattern; inspect morphology and controls, then seek membrane-associated signal in a reported positive population (UniProt Q15077 subcellular location; HPA tissue IHC; general IHC practice). |
| Q: Can the HPA ICC-IF entry confirm an IF pattern or supply an IF/ICC protocol? | HPA summarizes the location as Membrane but provides no main location or cell-line ICC-IF images in the supplied entry (HPA subcellular ICC-IF). | A: Use that entry only as limited localisation context alongside UniProt's cell-membrane annotation; assess IF/ICC on its separate guide page, since these payloads provide no IF/ICC protocol (HPA subcellular ICC-IF; UniProt Q15077 subcellular location). |
Experiment evidence
IHC & IF images showing P2RY6 staining
M10087-1 · IHCIHC analysis of P2Y6 using anti-P2Y6 antibody (M10087-1). P2Y6 was detected in a paraffin-embedded section of human colon cancer tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171631-P2RY6/tissue/cerebral+cortex#imid_20663147
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171631-P2RY6/tissue/colon#imid_20663057
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171631-P2RY6/tissue/liver#imid_20663074
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000171631-P2RY6/tissue/kidney#imid_20663067
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for P2RY6 IHC & IF
🧪Run tonsil first and score staining in its squamous epithelial cells (HPA: High in tonsil squamous epithelial cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the tonsil slide, use cells without discernible specific staining to judge local background, without assuming another cell type is P2RY6-negative.
Positive control tissue: Tonsil (Squamous epithelial 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 P2RY6; derive a cell-line control from the positive tissue's cell type (Squamous epithelial cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, a rabbit control immunoglobulin matched to the primary antibody’s clonality, and a P2RY6 knockout specimen if available (caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase and check the no-primary tonsil section for DAB background (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M10087-1 caption does not state a fixative (selected-SKU tissue-IHC caption). The caption uses heat retrieval in EDTA at pH 8.0 on a paraffin section; it does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU tissue-IHC caption). Endogenous peroxidase can produce chromogenic background, so assess it on the tonsil control slide (caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for P2RY6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Tonsil | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Appendix | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Goblet cells | Medium | Protein (IHC) | HPA → |
| Epididymis | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced P2RY6 IHC Tips
Troubleshoot P2RY6 staining in paraffin sections by checking retrieval, cellular context and membrane localisation before scoring chromogenic signal.
What retrieval should I start with for P2RY6 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M10087-1). The selected tissue image used this retrieval before incubation with the catalog antibody at 1:50 overnight at 4°C (datasheet M10087-1). If staining is weak, first check that sections were fully deparaffinised and compare retrieval duration across adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Record tissue preservation and background alongside signal, because excess heat can damage morphology and make staining harder to assign to individual cells (standard IHC practice).
How should I troubleshoot fixation-related loss of P2RY6 staining?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (datasheet M10087-1). Record each specimen’s fixative, fixation duration and processing history before comparing staining across batches (standard IHC practice). If signal varies, stain adjacent sections under the same EDTA pH 8.0 retrieval and 1:50 primary-antibody conditions used in the selected image, then examine morphology and staining together (datasheet M10087-1; standard IHC practice). Treat a fixation effect as a working hypothesis until a controlled comparison separates it from differences in retrieval, section quality or detection (standard IHC practice).
Should P2RY6 appear at the cell surface or throughout the cell?
P2RY6 is annotated at the cell membrane and has 7 transmembrane segments (UniProt Q15077 subcellular location and topology). In chromogenic IHC, inspect whether staining outlines cells or lies near their periphery, while recognising that DAB and section thickness may limit precise membrane assignment (UniProt Q15077 subcellular location; standard IHC practice). Compare the same cell population across a positive section, a negative control and adjacent morphology before accepting diffuse cytoplasmic colour (standard IHC practice). Strong nuclear-only staining conflicts with the annotated location and warrants checks of primary-antibody specificity, counterstain and detection background (UniProt Q15077 subcellular location; standard IHC practice).
Could the antibody epitope explain inconsistent P2RY6 staining?
The supplied record lists one 328-amino-acid chain, no annotated isoforms and an N-linked glycosylation site at residue 5 (UniProt Q15077 processing, isoforms and glycosylation). Its extracellular and cytoplasmic loops alternate around 7 transmembrane segments, so epitope location could affect accessibility after processing (UniProt Q15077 topology; standard IHC practice). The selected caption does not identify the antibody epitope, so neither loop accessibility nor glycosylation can be assigned as the cause of a staining difference (datasheet M10087-1). Compare retrieval and staining on serial sections, and seek documented epitope information before attributing discrepancies to a particular domain (standard IHC practice).
How can I use IF to investigate an ambiguous IHC pattern?
Use IF on a separate section to compare P2RY6 with a marker for the cell population implicated by the chromogenic pattern, such as an immune-cell marker in splenic red pulp (HPA: medium staining in cells of splenic red pulp; standard IF practice). Select spectrally separated fluorophores and inspect an unstained section for tissue autofluorescence before interpreting overlap (standard IF practice). Because the antibody’s epitope side is unspecified, test gentle permeabilisation alongside an unpermeabilised condition if membrane-side accessibility is in question (datasheet M10087-1; UniProt Q15077 topology; standard IF practice). Treat IF as a complementary localisation check, with its own staining controls, rather than transferring the paraffin-section IHC conditions directly (standard IF practice).
What should I check when DAB background obscures P2RY6 staining?
The selected image used a peroxidase-conjugated secondary antibody and DAB detection after 10% goat-serum blocking (datasheet M10087-1). Check a no-primary control, apply an appropriate endogenous-peroxidase block and inspect whether colour persists in the same structures without primary antibody (standard IHC practice). If background remains, compare shorter chromogen development, more thorough washes and a primary-antibody titration against the reported 1:50 condition while keeping retrieval fixed at EDTA pH 8.0 (datasheet M10087-1; standard IHC practice). Judge each adjustment against cell morphology and the expected membrane-associated distribution, since weaker overall colour alone does not establish specificity (UniProt Q15077 subcellular location; standard IHC practice).
How should I score P2RY6 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measuring P2RY6, because HPA reports staining in selected cell populations across tissues (HPA tissue IHC; standard IHC practice). For an identified population, report the percentage of positive cells and an intensity-based H-score from 0–300, using the same thresholds and exposure-independent viewing conditions across slides (standard IHC practice). If counting discrete positive cells, report density per mm² of viable tissue and the number of cells assessed, excluding folds, edges and necrotic regions (standard IHC practice). Normalise comparisons to the same annotated cell population or viable tissue area, with a batch control to track staining variation (standard IHC practice).
How can I distinguish true P2RY6 signal from artefact?
Prioritise staining associated with plausible cell membranes in an identified cell population, consistent with P2RY6’s annotated membrane location (UniProt Q15077 subcellular location; standard IHC practice). HPA reports high staining in tonsillar squamous epithelial cells and medium staining in appendiceal germinal-center cells, but its tissue reliability notes only medium consistency with RNA expression (HPA tissue IHC). Distrust isolated nuclear-only signal, staining confined to section edges or necrosis, and DAB deposits reproduced in a no-primary control (UniProt Q15077 subcellular location; standard IHC practice). Confirm a disputed pattern with morphology, a matched control and an independent specificity check before interpreting it as P2RY6 expression (standard IHC practice).
Boster reagents
Best P2RY6 / P2Y purinoceptor 6 IHC Antibodies
The catalog antibody has a human paraffin-section IHC image and lists IF/ICC among its applications; no IF image is supplied (M10087-1 catalog and IHC caption).
Real IHC data
Anti-P2Y6 Rabbit Monoclonal Antibody
Cat # M10087-1
M10087-1 is listed for human IHC and IF/ICC (M10087-1 catalog). Its IHC image shows staining in a paraffin-embedded human colon cancer section; no IF image is supplied (M10087-1 IHC caption and catalog).
Which to pick: Choose M10087-1 for paraffin-section IHC because its own caption documents that preparation; the fixative is unreported (M10087-1 IHC caption). For IF/ICC, M10087-1 is listed as applicable, but the payload supplies no IF image (M10087-1 catalog). Cross-species use is unsupported here because human is the only listed reactivity (M10087-1 catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15077 (P2RY6_HUMAN, P2Y purinoceptor 6).
- Human Protein Atlas. P2RY6 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. P2RY6 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. P2RY6 antibody validation summary (1 antibodies).
- Immunological role and clinical prognostic significance of P2RY6 in lung adenocarcinoma: a multi-omics studies and single-cell sequencing analysis. World journal of surgical oncology 2023 — PMC10601148.
- The microglial P2Y(6) receptor mediates neuronal loss and memory deficits in neurodegeneration. Cell reports 2021 — PMC8733854.
- lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window. International journal of molecular medicine 2019 — PMC6844642.
- P2RY6(+) tumor-associated macrophages as a therapeutic target: suppressing tumor growth and enhancing chemosensitivity in triple-negative breast cancer. Cellular oncology (Dordrecht, Netherlands) 2026 — PMC13473056.
- PubMed PMID:8670200 — UniProt-cited evidence.
- PubMed PMID:9286708 — UniProt-cited evidence.
- PubMed PMID:9268704 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for P2RY6 (P2Y purinoceptor 6). Expected localisation Cell membrane expected (UniProt); tissue compartment unassigned (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M10087-1). Validated IHC antibody: M10087-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Tonsil (Squamous epithelial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M10087-1); 2. Fixation: Image fixative and duration unreported (datasheet M10087-1); 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 retrieval: EDTA pH 8.0 (datasheet M10087-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M10087-1); 8. Primary antibody: Rabbit monoclonal (clone 19P56) anti-P2RY6, 1:50 (datasheet M10087-1); 9. Primary incubation: Overnight at 4 °C (datasheet M10087-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining in unannotated cells complicates assignment (HPA tissue IHC); Isoform / epitope: No isoforms or processing annotated; epitope side matters (UniProt); Regulation: Lymphoid tissue–enhanced RNA (HPA tissue RNA). Evidence: UniProt Q15077, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet M10087-1, PMC10601148, PMC6844642. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/P2RY6/immunohistochemistry-ihc-guide.