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CCL17 / C-C motif chemokine 17 · IHC design guide
Design Immunohistochemistry for CCL17
This guide uses the reported cytoplasmic tissue profile to plan chromogenic CCL17 IHC and select controls (HPA tissue IHC). The catalog antibody is validated for human IHC-P (datasheet: A02036); interpret staining with the uncertainty associated with a secreted protein in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCL17 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues and immune cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02036) | |
| Caveat | Secretion can separate tissue staining from RNA location (HPA tissue IHC) | |
| Regulation | Induced in stimulated PBMCs (UniProt) | |
| Isoform / epitope | No isoforms; 24–94 is secreted, with no cytoplasmic tail (UniProt) |
Section 1
Recommended CCL17 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published CCL17 IHC protocols for renal and lung tissue (PMC5299767; PMC3665443; PMC2741459).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02036); 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 anti-CCL17, 1:100–1:500 starting range (standard) |
| 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 | CCL17-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues and in immune cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page retrieval); the cited excerpts do not specify retrieval conditions.
Section 2
What Is the Expected CCL17 Staining Pattern?
CCL17 is a secreted chemokine with no transmembrane segment (UniProt Q92583). In tissue IHC, HPA reports predominantly cytoplasmic staining across many tissues and immune cells, including medium staining in appendix glandular cells and bone marrow hematopoietic cells (HPA tissue IHC). Treat this as a provisional pattern: HPA rates the tissue result and the listed antibody CAB002583 as Uncertain (HPA tissue IHC; HPA antibodies).
| Cytoplasmic staining in appendix glandular cells or bone marrow hematopoietic cells, around the reported medium level (HPA tissue IHC). | This resembles the observed CCL17 tissue pattern (HPA tissue IHC). Compare the stained cells with tissue architecture and a matched control section (general IHC practice). The match supports interpretation, but does not establish antibody specificity: the tissue result and CAB002583 IHC validation are Uncertain (HPA tissue IHC; HPA antibodies). |
| Strong, sharply nuclear or continuous cell-surface staining dominates, with little cytoplasmic signal. | A dominant nuclear or membrane pattern differs from HPA's cytoplasmic tissue profile (HPA tissue IHC). CCL17 is secreted and lacks a transmembrane segment, so continuous membrane staining is not the expected intrinsic localization (UniProt Q92583). Review controls and detection conditions before assigning this signal to CCL17 (general IHC practice). |
| Prominent staining appears in adipocytes or soft-tissue fibroblasts while expected cell types are weak (HPA tissue IHC). | HPA reports CCL17 as not detected in those cell types, making the result discordant with its tissue observations (HPA tissue IHC). Cross-reactivity or endogenous detection activity are possibilities to check with appropriate controls (general IHC practice). An HPA negative observation is not proof that expression is impossible (HPA tissue IHC: Uncertain). |
| Color spreads broadly across the section without a clear cell-associated pattern. | Diffuse background cannot be scored as the reported cytoplasmic cell staining (HPA tissue IHC). Because CCL17 is secreted, extracellular material may complicate localization, but diffuse color alone cannot identify its source (UniProt Q92583; general IHC practice). Check background controls, blocking, washes and detection conditions (general IHC practice). |
| No signal appears in appendix glandular cells or bone marrow hematopoietic cells (HPA tissue IHC). | Both are reported at medium intensity, so absence warrants a control and workflow review (HPA tissue IHC; general IHC practice). Check whether the section contains the relevant cells and whether detection worked (general IHC practice). These tissues are reference examples, not guaranteed positives, given the Uncertain HPA assessment (HPA tissue IHC). |
💡Expected CCL17 appearanceA provisional positive is predominantly medium cytoplasmic staining in appendix glandular or bone marrow hematopoietic cells (HPA tissue IHC); dominant nuclear staining or strong signal in HPA-undetected adipocytes is discordant and needs control review (HPA tissue IHC; general IHC practice).
| Secretion and compartment (UniProt Q92583; HPA tissue IHC) | CCL17 is secreted and has no transmembrane segment (UniProt Q92583), while HPA describes cytoplasmic tissue staining (HPA tissue IHC). Read cytoplasmic signal as an observed staining pattern, not proof that all CCL17 remains inside the producing cell (UniProt Q92583; HPA tissue IHC). |
| Processing and epitope interpretation (UniProt Q92583) | UniProt annotates a signal peptide at residues 1–23 and a mature chain at 24–94 (UniProt Q92583). The payload gives no antibody epitope, so peptide processing cannot predict whether CAB002583 detects precursor, mature secreted protein, or both (UniProt Q92583; HPA antibodies). |
| Tissue distribution and confidence (UniProt Q92583; HPA tissue IHC) | UniProt reports constitutive thymus expression and stimulated, but not resting, peripheral blood mononuclear cell expression (UniProt Q92583). HPA reports broad cytoplasmic tissue staining yet flags secreted-protein RNA/protein location differences and a transcript discrepancy (HPA tissue IHC). Interpret tissue comparisons with those limitations visible (HPA tissue IHC). |
| IHC antibody evidence (HPA antibodies; HPA tissue IHC) | The only listed antibody, CAB002583, has Uncertain IHC validation; the tissue profile is also Uncertain (HPA antibodies; HPA tissue IHC). A plausible compartment or tissue match alone therefore cannot settle specificity (HPA antibodies; HPA tissue IHC). |
| IF/ICC: what pattern is established? (HPA subcellular) | HPA calls CCL17 secreted but provides no main ICC/IF location or cell-line images (HPA subcellular). There is no image-backed intracellular IF pattern to prescribe here; assess any IF result against controls and the secreted nature of the target (HPA subcellular; UniProt Q92583; general IF practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in an HPA medium-positive tissue example (HPA tissue IHC). | The sampled section may lack the reported cells, or the IHC detection workflow may have failed (general IHC practice). | Confirm cell identity and tissue integrity, then inspect matched positive and detection controls before changing conditions (general IHC practice); HPA's examples remain provisional (HPA tissue IHC: Uncertain). |
| Signal is strong in adipocytes or soft-tissue fibroblasts (HPA tissue IHC). | These cells were not detected in the HPA profile; cross-reactivity or endogenous detection activity may contribute (HPA tissue IHC; general IHC practice). | Compare with a no-primary control and the expected cells in the same run; assess any residual detection signal before scoring CCL17 (general IHC practice). |
| A broad wash of chromogen obscures individual cells. | Background from blocking, washes or detection can obscure the reported cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Inspect the no-primary control and optimize general blocking, wash and detection conditions; score only interpretable cellular staining (general IHC practice). |
| The dominant signal is nuclear or forms a continuous membrane outline. | This differs from HPA's cytoplasmic tissue profile and from CCL17's secreted, non-transmembrane annotation (HPA tissue IHC; UniProt Q92583). | Check control sections and review antibody and detection specificity before assigning that compartment to CCL17 (general IHC practice). |
| Two tissue samples show different intensities. | Expression can vary by tissue and cell state: HPA lists medium and undetected examples, and UniProt distinguishes stimulated from resting blood mononuclear cells (HPA tissue IHC; UniProt Q92583). | Compare the same cell type and scoring criteria across sections; record the tissue context and retain HPA's Uncertain reliability caveat (general IHC practice; HPA tissue IHC). |
| An IF/ICC image appears to show a precise organelle location. | HPA provides no ICC/IF images or main intracellular location for CCL17; it describes the protein as secreted (HPA subcellular). | Treat the organelle assignment as unverified, check IF controls and report only the observed distribution until independent evidence supports a location (HPA subcellular; general IF practice). |
Experiment evidence
IHC & IF images showing CCL17 staining
A02036 · IHCImmunohistochemistry of CCL17 in human spleen tissue with CCL17 antibody at 5 μg/ml.
A02036 · IFImmunofluorescence of CCL17 in human spleen tissue with CCL17 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102970-CCL17/tissue/cerebral+cortex#imid_853760
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000102970-CCL17/tissue/stomach+2#imid_853644
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102970-CCL17/tissue/colon#imid_853678
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102970-CCL17/tissue/liver#imid_853679
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCL17 IHC & IF
🧪Run appendix first and look for staining in glandular cells (HPA: Medium in appendix glandular cells). Use adipose tissue as the negative, with adipocytes expected to lack detectable staining (HPA: Not detected in adipocytes); on the appendix slide, assess adjacent cells that lack signal as candidate internal negatives, without assuming their CCL17 status.
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CCL17; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, an immunoglobulin control matched to the primary antibody’s host species and class or monoclonal subclass, and CCL17 knockout tissue if available (standard IHC control practice). In appendix, check for endogenous peroxidase background, especially in inflammatory cells, and block it before chromogenic detection (standard IHC practice).
⚠️Feasibility: A CCL17-specific fixation window and retrieval dependence are unreported in the supplied evidence; the A02036 spleen IHC caption gives 5 µg/ml but leaves the fixative unreported (selected A02036 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier; secreted CCL17 can produce diffuse extracellular signal that complicates cell-level assignment (UniProt Q92583: Secreted). Interpret spleen staining cautiously because the caption shows spleen tissue while HPA reports no detectable signal in red-pulp cells (selected A02036 tissue-IHC caption; HPA: Not detected in spleen red-pulp cells).
HPA tissue IHC evidence for CCL17
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Secreted protein, tissue location of RNA and protein is expected to differ. Caution, Splice and/or transcript discrepancy exists.). 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 | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCL17 IHC Tips
Troubleshoot CCL17 chromogenic IHC by separating retrieval and staining problems from the interpretation of a secreted chemokine (UniProt Q92583).
What retrieval should I try first when CCL17 staining is weak?
Use citrate pH 6.0 heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval setting). Cool sections consistently before applying the IHC-validated antibody, and compare a retrieved section with a section processed without retrieval to assess whether heating improves signal (standard IHC practice). CCL17 is a secreted protein without a transmembrane segment, so a stained cell does not by itself establish where the protein was produced (UniProt Q92583). If signal remains weak, adjust retrieval time on matched sections while keeping detection and exposure to chromogen constant; excessive heating can damage morphology and complicate interpretation (standard IHC practice).
How can I assess whether fixation is suppressing CCL17 signal?
Target-specific fixation sensitivity is unknown: the selected spleen IHC caption reports staining at 5 µg/ml but does not state the fixative (caption A02036). Record the fixative and fixation interval for each specimen, then compare sections with similar processing histories before attributing a weak result to antibody performance (standard IHC practice). If material permits, compare fixation conditions using the same citrate pH 6.0 retrieval and chromogenic detection settings throughout (page retrieval setting; standard IHC practice). Evaluate tissue morphology alongside staining, because processing differences can alter both antigen accessibility and the apparent distribution of a secreted protein (standard IHC practice; UniProt Q92583).
Should CCL17 appear inside cells or in extracellular tissue?
Interpret cytoplasmic staining as plausible, because tissue IHC reports cytoplasmic expression in many tissues and immune cells (HPA tissue IHC). CCL17 is secreted and has no transmembrane segment, so extracellular or nearby cell-associated staining may reflect released protein rather than synthesis by the stained cell (UniProt Q92583). Compare cellular signal with adjacent extracellular spaces and tissue architecture, using a matched negative control to judge whether diffuse color exceeds nonspecific deposition (standard IHC practice). Avoid assigning a producer cell from chromogenic localisation alone; correlate the stained region with cell identity and, when available, an independent expression readout (UniProt Q92583; standard IHC practice).
Could a processed CCL17 epitope explain inconsistent tissue staining?
The 94-aa CCL17 precursor contains a signal peptide at residues 1–23, with the annotated mature chain spanning residues 24–94 (UniProt Q92583). An antibody recognising the signal peptide could behave differently from one recognising the mature chain, but the supplied caption does not identify the antibody epitope (UniProt Q92583; caption A02036). Check the catalog antibody’s immunogen or epitope documentation before assigning a processing-based explanation, and compare matched positive and negative staining controls (standard IHC practice). No alternative isoforms, glycosylation sites or modified residues are annotated here, so avoid invoking any of them to explain an unexpected pattern (UniProt Q92583).
How should I adapt the interpretation for multiplex CCL17 immunofluorescence?
For the separate IF/ICC workflow, pair CCL17 with a marker for the expected cell population, such as CD45 when testing immune-cell association, and assess overlap cell by cell (HPA tissue IHC: immune-cell expression; standard IF practice). Choose a far-red fluorophore when the specimen has strong shorter-wavelength autofluorescence, and include single-color controls to check bleed-through (standard IF practice). For an intracellular epitope, use a mild permeabilisation condition; for accessible extracellular CCL17, compare with an unpermeabilised preparation because detergent may change retained signal (UniProt Q92583: secreted; standard IF practice). No ICC/IF images are supplied here, so verify the antibody’s IF performance independently of its IHC caption (HPA subcellular; caption A02036).
How do I distinguish CCL17 signal from chromogenic background?
Inspect a no-primary control processed through the same secondary reagent, peroxidase block and DAB development; color in that control indicates a detection-related contribution (standard IHC practice). Keep the blocking reagent, wash schedule and chromogen development time consistent across comparison sections, then change one variable at a time if diffuse staining persists (standard IHC practice). The spleen image caption gives an antibody concentration of 5 µg/ml but does not establish an optimal concentration for every specimen (caption A02036). Because CCL17 is secreted, extracellular color requires particular scrutiny against the no-primary control and surrounding morphology before it is called specific (UniProt Q92583; standard IHC practice).
What is a defensible way to quantify CCL17 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and scoring threshold before reviewing groups, then record the percentage of positive cells and their staining intensity on matched sections (standard IHC practice). An H-score combines intensity grades 0–3 with the percentage at each grade, giving a range of 0–300; report the scoring rules and observer agreement (standard IHC practice). For immune-cell-rich regions, positive-cell density per mm² can complement H-score, provided counts are normalised to viable analysed area or the relevant cell population (standard IHC practice). Score extracellular deposition separately from cellular staining, because secreted CCL17 may occur away from its producer (UniProt Q92583).
Which CCL17 staining patterns warrant extra validation before calling a result positive?
Give greatest weight to reproducible staining with preserved morphology and a clean no-primary control, and scrutinise edge staining, necrotic regions and residual endogenous peroxidase activity (standard IHC practice). Strong nuclear-only staining warrants extra validation because CCL17 is annotated as secreted and tissue IHC describes a cytoplasmic pattern (UniProt Q92583; HPA tissue IHC). Do not treat a stained spleen section as universal positive-control evidence: the selected caption shows spleen staining, whereas HPA reports red-pulp cells as not detected and rates its tissue evidence uncertain (caption A02036; HPA tissue IHC). Confirm disputed patterns with an independent antibody or expression assay and document the exact cell population assessed (standard IHC practice).
Boster reagents
Best CCL17 / C-C motif chemokine 17 IHC Antibodies
A02036 has human spleen IHC and IF images (catalog image captions); its listed reactivity includes human, mouse, and rat (catalog reactivity).
Real IHC data
Anti-C-C motif chemokine 17 CCL17 Antibody
Cat # A02036
A02036 has an IHC image of human spleen at 5 μg/ml (catalog IHC caption). A02036 also has an IF image of human spleen at 20 μg/ml (catalog IF caption).
Which to pick: For tissue IHC, choose A02036: IHC-P is listed (catalog applications), and its IHC image shows human spleen at 5 μg/ml (catalog IHC caption); the caption does not report the fixative (catalog IHC caption). For IF, choose A02036 based on its human spleen image (catalog IF caption); ICC validation is unreported (catalog applications). For mouse or rat work, A02036 lists both species (catalog reactivity), but the supplied IHC and IF images show human tissue only (catalog image captions); it is rabbit hosted, with clonality unreported (catalog payload).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92583 (CCL17_HUMAN, C-C motif chemokine 17).
- Human Protein Atlas. CCL17 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. CCL17 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. CCL17 antibody validation summary (1 antibodies).
- Low CCL17 expression associates with unfavorable postoperative prognosis of patients with clear cell renal cell carcinoma. BMC cancer 2017 — PMC5299767.
- Higher serum CCL17 may be a promising predictor of acute exacerbations in chronic hypersensitivity pneumonitis. Respiratory research 2013 — PMC3665443.
- Macrophage derived chemokine (CCL22), thymus and activation-regulated chemokine (CCL17), and CCR4 in idiopathic pulmonary fibrosis. Respiratory research 2009 — PMC2741459.
- NOTCH1 fusions in pediatric T-cell lymphoblastic lymphoma: A high-risk subgroup with CCL17 (TARC) levels as diagnostic biomarker. HemaSphere 2024 — PMC11208779.
- PubMed PMID:8702936 — UniProt-cited evidence.
- PubMed PMID:10493829 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCL17 (C-C motif chemokine 17). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02036 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A02036); 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 anti-CCL17, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02036); Caveat: Secretion can separate tissue staining from RNA location (HPA tissue IHC); Isoform / epitope: No isoforms; 24–94 is secreted, with no cytoplasmic tail (UniProt); Regulation: Induced in stimulated PBMCs (UniProt). Evidence: UniProt Q92583, HPA tissue IHC · Uncertain, HPA ICC-IF · Secreted, Datasheet A02036, PMC5299767, PMC3665443. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCL17/immunohistochemistry-ihc-guide.