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CCR3 / C-C chemokine receptor type 3 · IHC design guide
Design Immunohistochemistry for CCR3
Plan CCR3 paraffin-section IHC around the mainly membranous tissue pattern reported by HPA (HPA tissue IHC). Compare controls and interpret staining cautiously because HPA rates its tissue staining reliability as uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCR3 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 membranous in tissue (HPA tissue IHC) | |
| Staining pattern | Mainly membranous staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining; reliability uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended CCR3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by three published CCR3 immunohistochemistry workflows: frozen mouse brain, paraffin-embedded bovine corpus luteum, and human prostate cancer tissue microarrays.
| Sample | Paraffin-embedded human epidemal cells of skin tissue; fixative not specified (datasheet M01748) |
| Fixation | Image fixative and duration unreported (datasheet M01748); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit monoclonal (clone ECE-3) anti-CCR3, 1:50 (datasheet M01748) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCR3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Mainly membranous expression in several tissues. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule).
Section 2
What Is the Expected CCR3 Staining Pattern?
CCR3 is a seven-transmembrane receptor assigned to the cell membrane, so a credible positive cell should show a membranous signal (UniProt P51677 topology; UniProt P51677 subcellular location). Eosinophils are the clearest cell-type expectation; UniProt reports only trace amounts in neutrophils and monocytes (UniProt P51677 tissue specificity). HPA describes mainly membranous tissue staining, but rates its tissue IHC profile Uncertain because staining and RNA data have low consistency and presumed off-target binding was observed (HPA tissue IHC).
| A crisp membrane-associated signal appears in cells identified as eosinophils by morphology. | This fits the receptor's location and strongest stated cell-type expectation (UniProt P51677 subcellular location and tissue specificity). Judge the signal alongside the tissue context and controls; HPA's overall tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Signal is predominantly nuclear or fills the cytoplasm without a discernible membrane component. | That distribution conflicts with the assigned cell-membrane location and seven-transmembrane topology (UniProt P51677 subcellular location and topology). Treat it as suspect and check detection background before calling the cell positive (general IHC practice). |
| Strong staining appears in an unexpected cell population, especially across many cell types. | Investigate nonspecific binding or endogenous detection activity (general IHC practice). Cell identity alone is not decisive: HPA reports medium staining in several epithelial or glandular populations, while rating its tissue IHC profile Uncertain (HPA tissue IHC). |
| A diffuse chromogenic haze obscures cell borders throughout the section. | This cannot establish the membrane pattern expected for CCR3 (UniProt P51677 subcellular location; HPA tissue IHC profile). Compare a no-primary control, then review blocking, washes and detection conditions (general IHC practice). |
| There is no signal in a specimen with independently established CCR3-positive cells. | First confirm that those cells are present and that the detection controls worked (general IHC practice). HPA tissue examples alone are a weak positive-control standard because its tissue IHC reliability is Uncertain (HPA tissue IHC). |
💡Expected CCR3 appearanceCall a result positive when identifiable eosinophils show distinct membrane-associated chromogen above local background; intensity may vary, while isolated diffuse or nuclear color is suspect (UniProt P51677 tissue specificity and subcellular location; HPA tissue IHC profile; general IHC practice).
| Topology and compartment | CCR3 has seven transmembrane segments and is assigned to the cell membrane (UniProt P51677 topology and subcellular location). Score a membrane-associated pattern rather than using total cellular color as the sole criterion (general IHC practice). |
| Cell population | UniProt places CCR3 in eosinophils and reports trace amounts in neutrophils and monocytes (UniProt P51677 tissue specificity). Cell morphology therefore matters when interpreting a stained inflammatory infiltrate (general IHC practice). |
| Tissue-level evidence | HPA reports mainly membranous staining and examples of medium epithelial or glandular staining, but flags low agreement with RNA expression and presumed off-target binding (HPA tissue IHC). Its tissue positives should be interpreted cautiously. |
| Antibody validation | The listed tissue IHC antibody CAB003795 is rated Uncertain; the separate antibody HPA069514 has Supported ICC evidence but no listed IHC rating (HPA antibodies). ICC support does not establish IHC-P performance for another antibody (general assay interpretation). |
| IF/ICC applicability | For the separate IF/ICC guide: HPA assigns an approved plasma-membrane location and lists images from RT-4 and U2OS (HPA subcellular ICC-IF). This location supports an IF interpretation, without changing the Uncertain tissue IHC assessment (HPA tissue IHC). |
| Isoforms and epitope coverage | UniProt lists two CCR3 isoforms (UniProt P51677 isoforms). The supplied record does not establish which isoform an IHC antibody recognizes; check the antibody's stated epitope before interpreting a negative result as absence of all CCR3 isoforms (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No chromogen appears in an independently confirmed positive control. | A failed detection step or unsuitable assay conditions are possibilities (general IHC practice); HPA tissue examples alone do not resolve this because their IHC reliability is Uncertain (HPA tissue IHC). | Verify the control contains the expected cells, check the detection reagents, and review the IHC-P antibody instructions, including any specified retrieval and dilution (general IHC practice). |
| Staining is mostly nuclear or uniformly cytoplasmic. | The pattern disagrees with CCR3's assigned membrane location (UniProt P51677 subcellular location). Background or nonspecific staining may be contributing (general IHC practice). | Compare no-primary and appropriate tissue controls; assess whether any clear membrane-associated signal remains after background is accounted for (general IHC practice). |
| Many unrelated cell populations stain at similar strength. | Nonspecific binding or endogenous detection activity may contribute (general IHC practice). HPA also reports presumed off-target binding in its uncertain tissue IHC assessment (HPA tissue IHC). | Inspect no-primary controls, review blocking and detection chemistry, and require a plausible cellular distribution before scoring the section (general IHC practice; UniProt P51677 tissue specificity). |
| The whole section has diffuse color and cell borders are hard to resolve. | Background can obscure a membrane-associated result (general IHC practice; UniProt P51677 subcellular location). | Check no-primary staining, wash performance and detection conditions; score only cells whose membrane pattern can be distinguished from local background (general IHC practice). |
| An epithelial population stains, but eosinophils are not identified. | HPA records some medium epithelial staining, yet rates the overall tissue IHC profile Uncertain (HPA tissue IHC). The observation alone cannot confirm cell-specific CCR3 expression. | Identify cells by morphology, compare suitable controls, and report the epithelial pattern as an observation rather than assigning CCR3 specificity from color alone (general IHC practice; HPA tissue IHC). |
| IF/ICC shows a membrane signal while paraffin IHC is inconclusive. | HPA's approved plasma-membrane ICC-IF location and its Uncertain tissue IHC rating are distinct assessments (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret each application with its own controls and antibody validation; do not transfer an ICC result to the IHC-P interpretation (HPA antibodies; general assay interpretation). |
Experiment evidence
IHC & IF images showing CCR3 staining
M01748 · IHCImmunohistochemical analysis of paraffin-embedded human epidemal cells of skin, using CCR3 Antibody .
M01748 · IHCImmunohistochemical analysis of paraffin-embedded human epidemal cells of skin, using CCR3 Antibody .
PA2176 · IHCAnti-CCR3 antibody, PA2176, IHC(P) IHC(P): Human Tonsil Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000183625-CCR3/tissue/cerebral+cortex#imid_1252740
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000183625-CCR3/tissue/esophagus#imid_1252728
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000183625-CCR3/tissue/colon#imid_1252674
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000183625-CCR3/tissue/liver#imid_1252667
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA069514; green = CCR3, blue = nucleus, red = microtubules.
HPA · ICC-IF RT-4Human Protein Atlas immunofluorescence, RT-4, antibody HPA069514; green = CCR3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCR3 IHC & IF
🧪Run esophagus first: squamous epithelial cells should stain (HPA: Medium in esophageal squamous epithelial cells). Use adipose tissue as the negative comparator, where adipocytes are not detected (HPA: Not detected in adipocytes); on the positive slide, score cells with no specific membrane staining as internal negatives without assuming every nonepithelial cell is CCR3-negative (UniProt P51677: cell membrane; eosinophil expression).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCR3 in RT-4, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include no-primary or secondary-only and host-species- and clonality-matched isotype controls, plus CCR3 knockout tissue as a biological negative (standard IHC practice). Quench endogenous peroxidase for chromogenic detection and check the esophagus slide for background staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; test retrieval conditions on matched sections (standard IHC practice). The selected M01748 paraffin-section caption does not report a fixative, so it does not establish fixation (datasheet: M01748 tissue-IHC caption). IF/ICC has plasma-membrane imaging evidence in RT-4 and U2OS (HPA subcellular), but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC; assess background staining in esophageal epithelium during scoring (standard IHC practice). The selected M01748 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image M01748).
HPA tissue IHC evidence for CCR3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCR3 IHC Tips
Troubleshoot CCR3 chromogenic IHC in paraffin sections by checking retrieval, compartment, cell identity and controls before interpreting staining.
How should I retrieve CCR3 in paraffin sections when membrane staining is weak?
Use citrate pH 6.0 HIER at 95–98 °C for 20 min for CCR3 paraffin IHC (page retrieval rule). Allow sections to cool in retrieval buffer, then compare staining with a matched no-primary control processed through the same steps (standard IHC practice). CCR3 has 7 transmembrane segments, and the expected signal is membranous, so assess cell borders rather than overall brown intensity (UniProt P51677 topology; HPA subcellular). If staining remains weak, pilot a different retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Record any increase in diffuse background alongside the change in membrane signal (standard IHC practice).
Could fixation explain absent CCR3 staining in my paraffin sections?
Target-specific CCR3 sensitivity to fixation is unknown from the supplied evidence, so fixation cannot be diagnosed as the cause of a negative section (supplied fixation evidence). The selected M01748 image describes paraffin-embedded human epidermal cells of skin but does not state a fixative (M01748 IHC caption). Record the fixative, fixation duration and tissue processing history for each specimen, and compare sections processed together before changing antibody conditions (standard IHC practice). If fixation varies between specimens, run retrieval and staining on adjacent sections in the same batch, with a no-primary control for each condition (standard IHC practice). Interpret any recovery as a workflow observation, not proof of CCR3-specific fixation sensitivity (standard IHC practice).
What staining pattern should count as CCR3 localisation in tissue IHC?
Prioritise staining along cell borders: CCR3 is annotated at the cell membrane, and HPA reports mainly membranous tissue expression (UniProt P51677 subcellular; HPA tissue IHC profile). Its 7 transmembrane segments provide a structural reason to inspect membrane contours in intact cells (UniProt P51677 topology). Score cytoplasmic haze separately, and compare both patterns with the no-primary control and neighboring tissue regions (standard IHC practice). HPA approves a plasma-membrane location from subcellular imaging, but rates its tissue IHC reliability Uncertain because staining and RNA data show low consistency and presumed off-target binding (HPA subcellular; HPA tissue IHC reliability). Therefore, require reproducible cell-border staining before assigning a tissue-level positive result (standard IHC practice).
How does epitope location affect interpretation of CCR3 staining?
Check the catalog antibody's stated immunogen or epitope before choosing controls; the supplied record does not locate its binding site (supplied antibody evidence). CCR3 has extracellular residues 1–34 and 265–281, plus a cytoplasmic tail at 306–355, so accessibility depends on which region the antibody recognizes (UniProt P51677 topology). The record lists 2 isoforms but supplies no isoform-specific epitope map, so a positive IHC result cannot identify one isoform by itself (UniProt P51677 isoforms; supplied epitope evidence). Compare staining across adjacent sections under identical retrieval and detection conditions, and document any epitope information obtained before interpreting discordant signals (standard IHC practice).
How can IF help assess a questionable CCR3 IHC signal?
On a separately optimised IF preparation, multiplex CCR3 with an eosinophil marker to check the expected cell population; UniProt reports CCR3 in eosinophils and trace amounts in neutrophils and monocytes (UniProt P51677 tissue specificity). Choose a fluorophore channel after measuring tissue autofluorescence with an unstained control, and include single-colour controls when separating overlapping signals (standard IF practice). Match permeabilisation to the antibody epitope: an extracellular epitope can be assessed without permeabilisation, whereas a cytoplasmic epitope requires access across the membrane (UniProt P51677 topology; standard IF practice). HPA reports an approved plasma-membrane location in subcellular imaging, but its tissue IHC reliability is Uncertain; agreement between methods supports localisation without proving tissue specificity (HPA subcellular; HPA tissue IHC reliability).
What should I check when CCR3 IHC has diffuse brown background?
Compare the section with a no-primary control and inspect whether staining follows tissue edges, damaged areas or intact cell borders (standard IHC practice). For chromogenic detection, block endogenous peroxidase according to the detection workflow and check whether the DAB reaction was stopped consistently across slides (standard IHC practice). Titrate the catalog antibody and detection reagent on serial sections while keeping citrate pH 6.0 retrieval at 95–98 °C for 20 min constant (page retrieval rule; standard IHC practice). CCR3 is a membrane receptor, so diffuse staining without a convincing border pattern needs particular scrutiny (UniProt P51677 subcellular). HPA flags presumed off-target tissue staining, making matched controls essential before calling weak signal positive (HPA tissue IHC reliability).
How should I quantify CCR3-positive cells across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis compartment as intact cell-border staining before scoring, consistent with CCR3 membrane localisation (UniProt P51677 subcellular; standard IHC practice). For a comparable cell population, report the percentage of positive cells and an H-score using intensity grades 0–3, giving a possible range of 0–300 (standard IHC practice). If the question concerns infiltrating cells, count positive cells per mm² of viable tissue and normalise to tissue area or the total number of eligible cells (standard IHC practice). Apply the same threshold, section thickness and imaging settings across specimens, and record excluded edge or necrotic regions (standard IHC practice). Report membrane and cytoplasmic scores separately because HPA tissue IHC reliability is Uncertain (HPA tissue IHC reliability).
When is a CCR3-positive IHC result biologically credible?
A credible signal traces intact cell membranes and is reproducible in comparable sections, matching the annotated CCR3 location (UniProt P51677 subcellular; standard IHC practice). Check cell identity with morphology or a suitable companion marker: CCR3 is reported in eosinophils, with trace expression in neutrophils and monocytes (UniProt P51677 tissue specificity). Reject isolated edge staining, necrotic deposits or colour reproduced in the no-primary control, and check endogenous peroxidase when brown precipitate is widespread (standard IHC practice). Do not treat epithelial staining alone as confirmation: HPA lists some epithelial positives but assigns tissue IHC Uncertain reliability because of low RNA concordance and presumed off-target binding (HPA tissue IHC positive entries; HPA tissue IHC reliability).
Boster reagents
Best CCR3 / C-C chemokine receptor type 3 IHC Antibodies
Both catalog antibodies have human tissue IHC images: paraffin-embedded skin cells for M01748 and tonsil for PA2176 (respective IHC captions). M01748 also lists mouse/rat reactivity and IF/ICC applications (M01748 catalog).
Real IHC data
Anti-CCR3 Rabbit Monoclonal Antibody
Cat # M01748
Real IHC data
Anti-CCR3 Antibody ®
Cat # PA2176
M01748 renders with its paraffin-embedded human skin-cell IHC image and lists IHC, ICC and IF applications with human, mouse and rat reactivity (M01748 catalog; IHC caption). PA2176 renders with its human tonsil IHC(P) image and lists IHC with human reactivity (PA2176 catalog; IHC caption).
Which to pick: For tissue IHC, choose PA2176 if its human tonsil IHC(P) example matches your sample, or M01748 if its paraffin-embedded human skin example is more relevant (respective IHC captions). For IF/ICC or mouse/rat samples, M01748 is the listed option: it is rabbit monoclonal clone ECE-3 and lists those applications and species (M01748 catalog). The M01748 caption reports paraffin embedding, while the PA2176 caption reports IHC(P); neither reports the fixative (respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51677 (CCR3_HUMAN, C-C chemokine receptor type 3).
- Human Protein Atlas. CCR3 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. CCR3 subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. CCR3 antibody validation summary (2 antibodies).
- Exacerbation of facial motoneuron loss after facial nerve axotomy in CCR3-deficient mice. ASN neuro 2009 — PMC2826103.
- Chemokine Receptor Profiles as Predictors of Survival and Early Progression in Follicular Lymphoma. EJHaem 2025 — PMC12393056.
- Gene expression profiles in the bovine corpus luteum (CL) during the estrous cycle and pregnancy: possible roles of chemokines in regulating CL function during pregnancy. The Journal of reproduction and development 2015 — PMC4354230.
- Periprostatic adipocytes act as a driving force for prostate cancer progression in obesity. Nature communications 2016 — PMC4729927.
- PubMed PMID:7622448 — UniProt-cited evidence.
- PubMed PMID:8642344 — UniProt-cited evidence.
- PubMed PMID:8676064 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCR3 (C-C chemokine receptor type 3). Expected localisation Mainly membranous in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M01748 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human epidemal cells of skin tissue; fixative not specified (datasheet M01748); 2. Fixation: Image fixative and duration unreported (datasheet M01748); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit monoclonal (clone ECE-3) anti-CCR3, 1:50 (datasheet M01748); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining; reliability uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unknown (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P51677, HPA tissue IHC · Uncertain, HPA ICC-IF · plasma membrane., Datasheet M01748, PMC2826103, PMC4354230. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCR3/immunohistochemistry-ihc-guide.