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CCL17 / C-C motif chemokine 17 · IHC design guide
Design Immunohistochemistry for CCL17
Plan chromogenic CCL17 IHC in paraffin sections using the antibody validated in human IHC (datasheet). Assess cytoplasmic staining with matched controls, bearing in mind that CCL17 is secreted and tissue RNA and protein locations may differ (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 staining in most tissues and immune cells (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in glandular 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 | Secreted protein: tissue RNA and protein sites may differ (HPA tissue IHC) | |
| Regulation | Stimulated PBMCs only; not resting (UniProt) | |
| Isoform / epitope | No isoforms; mature chain 24–94 follows signal cleavage (UniProt) |
Section 1
Recommended CCL17 IHC & IF Protocols
The catalog antibody protocol is followed by published CCL17 IHC methods for renal tumor microarrays and lung sections (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 retrieval at 95–98 °C for 20 min (page retrieval rule); optimize staining against tissue controls.
Section 2
What Is the Expected CCL17 Staining Pattern?
CCL17 is secreted with no transmembrane segment (UniProt Q92583); cytoplasmic immune and glandular staining is reported (HPA: Uncertain).
| Medium cytoplasmic stain in appendix glands | Matches HPA: Medium glandular staining; IHC reliability is Uncertain. |
| Nuclear-only staining | Conflicts with the HPA cytoplasmic profile; investigate artefact. |
| Staining in adipocytes | HPA: Not detected; consider cross-reactivity or endogenous detection activity. |
| Diffuse staining across the section | May reflect background; check blocking and detection controls (general IHC practice). |
| No stain in appendix glands | Unexpected versus HPA: Medium; review the positive control and detection conditions. |
💡Expected CCL17 appearanceExpect medium cytoplasmic appendix gland staining (HPA: Medium); nuclear-only stain is suspect (HPA: cytoplasmic).
| Secretion and topology | Secreted with no transmembrane segment; extracellular signal is possible (UniProt Q92583). |
| Protein processing | Signal peptide is residues 1–23; mature chain is 24–94 (UniProt Q92583). |
| Tissue selection | Appendix glands stain Medium; adipocytes are Not detected (HPA). |
| Antibody validation | CAB002583 has Uncertain IHC reliability; interpret staining with controls (HPA). |
| IF/ICC: where should signal appear? | No main location or ICC-IF images are available (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No appendix gland signal | Unexpected (HPA: Medium). | Check the positive control, retrieval and detection (general IHC practice). |
| Adipocyte staining | Discordant (HPA: Not detected). | Check a no-primary control for detection background (general IHC practice). |
| Nuclear-only signal | Discordant (HPA: cytoplasmic). | Review counterstain and antibody specificity (general IHC practice). |
| Diffuse background | Nonspecific binding or endogenous enzyme activity (general IHC practice). | Optimize blocking and check a no-primary control (general IHC practice). |
| Extracellular signal dominates | CCL17 is secreted (UniProt Q92583). | Score cell-associated staining separately (general IHC 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 (Medium; HPA: appendix glandular cells). Use esophagus as the negative tissue, focusing on squamous epithelial cells (HPA: esophagus squamous epithelial cells Not detected); nonglandular cells on the appendix slide should provide a background comparison, but should not be assumed target-negative.
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 and a control immunoglobulin matched to the primary antibody’s host species and, for a monoclonal antibody, its isotype (standard IHC practice). Use CCL17 knockout tissue or a validated peptide block as a biological specificity control; check for endogenous peroxidase signal from inflammatory cells in appendix before interpreting chromogen staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected A02036 spleen IHC caption also leaves its fixative unreported (catalog caption: A02036). The evidence does not establish that frozen sections or IF are easier than paraffin IHC, and no ICC-IF image cell lines are listed (HPA: CCL17 subcellular record). Because CCL17 is secreted (UniProt Q92583: Secreted), assess extracellular or luminal appendix signal cautiously when assigning staining to glandular 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 staining by checking retrieval, controls, cell identity, and the distinction between intracellular production and secreted protein (UniProt Q92583).
How should I optimize antigen retrieval when CCL17 staining is weak?
Start with citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min (this page’s retrieval rule). Allow sections to cool in buffer, then compare matched positive and negative controls processed in the same run (standard IHC practice). If staining remains weak, vary heating time in small increments while watching for tissue damage and rising background (standard IHC practice). Record the actual temperature reached and compare staining at a fixed antibody concentration, since simultaneous changes obscure the cause of improvement (standard IHC practice). Interpret weak cellular staining cautiously because CCL17 is secreted (UniProt Q92583 subcellular location).
Could fixation explain weak or uneven CCL17 staining in paraffin sections?
The selected tissue-IHC caption does not state a fixative, so CCL17-specific sensitivity to fixation is unknown (A02036 tissue-IHC caption). Document fixation conditions and compare sections with similar processing histories before changing antibody concentration or retrieval (standard IHC practice). For uneven staining, inspect tissue edges, folds, and poorly preserved areas against an intact region on the same section (standard IHC practice). Run a matched control section through the same citrate pH 6.0, 95–98 °C, 20 min retrieval to isolate processing differences (this page’s retrieval rule). Do not assign a fixation effect to CCL17 solely from staining distribution (standard IHC interpretation).
Should CCL17 appear in the cytoplasm, extracellular space, or both?
Prioritize interpretable cytoplasmic staining in intact cells, while recognizing that released CCL17 can appear beyond its producing cell (HPA tissue IHC profile; UniProt Q92583 subcellular location). CCL17 has a signal peptide at residues 1–23 and a mature chain at 24–94, consistent with secretion (UniProt Q92583 processing). It has no annotated transmembrane segment, so a crisp, exclusively membrane-bound pattern needs independent validation (UniProt Q92583 topology). Compare cellular signal with extracellular deposits on serial sections and note whether deposits track damaged or necrotic areas (standard IHC practice). HPA describes cytoplasmic expression in many tissues but rates its tissue-IHC reliability uncertain (HPA tissue IHC).
How do processing and epitope position affect CCL17 IHC interpretation?
Check whether the antibody’s documented immunogen or mapped epitope lies in the signal peptide or mature chain before interpreting staining (standard IHC practice; UniProt Q92583 processing). The signal peptide spans residues 1–23, whereas the annotated CCL17 chain spans 24–94 (UniProt Q92583 processing). An epitope confined to the cleaved signal peptide would not establish detection of secreted mature CCL17 (UniProt Q92583 processing). No isoforms, glycosylation sites, or modified residues are annotated in the supplied record, so these do not explain a pattern by themselves (UniProt Q92583 record). Confirm specificity with an appropriate independent control, especially where extracellular staining dominates (standard IHC practice).
How should I adapt CCL17 staining for multiplex immunofluorescence?
Treat IF/ICC as a separate optimization because the supplied A02036 image documents tissue IHC, with fixative unreported (A02036 tissue-IHC caption). Pair CCL17 with a validated marker for the suspected producing cell type and assess co-localization at the single-cell level (standard IF practice; HPA tissue IHC profile). Choose fluorophores after examining unstained tissue autofluorescence, and reserve a spectrally distinct channel for weak CCL17 signal (standard IF practice). If the antibody recognizes intracellular CCL17, titrate mild permeabilisation after fixation; a purely extracellular epitope can be tested without it (standard IF practice; UniProt Q92583 subcellular location). Include single-stain and secondary-only controls when evaluating multiplex signal (standard IF practice).
How can I distinguish CCL17 signal from diffuse chromogenic background?
Compare the test section with a matched no-primary control and inspect whether color follows intact cells or spreads across the section (standard IHC practice). Block endogenous peroxidase before chromogenic detection and check whether residual color persists in the no-primary control (standard IHC practice). Titrate the primary antibody and shorten chromogen development if both expected staining and background increase together (standard IHC practice). The selected caption reports 5 µg/ml in human spleen, but supplies no fixative or broader concentration range (A02036 tissue-IHC caption). Because CCL17 is secreted, diffuse extracellular color alone requires stronger specificity evidence than well-resolved cellular staining (UniProt Q92583 subcellular location; standard IHC interpretation).
What should I score when comparing CCL17 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring, and report cellular and extracellular signal separately (standard IHC practice; UniProt Q92583 subcellular location). For discrete positive cells, record % positive cells or positive-cell density per mm² of viable tissue (standard IHC quantification). If intensity varies reproducibly, calculate an H-score from the percentages of cells at each prespecified intensity grade (standard IHC quantification). Normalize counts to the relevant viable tissue area or eligible cell count, excluding folds, necrosis, and section edges by a consistent rule (standard IHC practice). Keep retrieval, detection, imaging, and scoring thresholds consistent across comparison groups (standard IHC practice).
When is an apparent CCL17-positive IHC result convincing?
Look for reproducible signal in intact cells with a plausible cytoplasmic pattern, supported by a low-background control (HPA tissue IHC profile; standard IHC practice). Treat isolated nuclear or sharply membrane-restricted staining cautiously because CCL17 is secreted and lacks an annotated transmembrane segment (UniProt Q92583 subcellular location and topology). Exclude edge effects, necrotic deposits, and endogenous enzyme signal by reviewing morphology and a matched no-primary control (standard IHC practice). The A02036 caption shows human spleen at 5 µg/ml, while HPA reports no detection in spleen red-pulp cells; those observations concern different sampling and assessment contexts (A02036 tissue-IHC caption; HPA tissue IHC). HPA rates its tissue-IHC reliability uncertain, so corroborate disputed cell assignments independently (HPA tissue IHC).
Boster reagents
Best CCL17 / C-C motif chemokine 17 IHC Antibodies
A02036 has IHC and IF images of human spleen; its application list includes IHC-P and IF, and its reactivity list includes human, mouse and rat (catalog applications, reactivity and image captions).
Real IHC data
Anti-C-C motif chemokine 17 CCL17 Antibody
Cat # A02036
A02036 will render with an IHC image of human spleen stained at 5 μg/ml (IHC image caption). Its IF image shows human spleen stained at 20 μg/ml; the catalog lists IHC-P and IF applications and human, mouse and rat reactivity (IF image caption; catalog applications and reactivity).
Which to pick: Choose A02036 for paraffin-section IHC because IHC-P is listed, with a human spleen IHC image at 5 μg/ml (catalog applications; IHC image caption). For IF, A02036 has a human spleen image at 20 μg/ml; ICC validation and clonality are unreported (IF image caption; catalog payload). For other species, the catalog lists mouse and rat reactivity, but its IHC and IF images show human spleen; the IHC image caption does not report the fixative or tissue processing (catalog reactivity; IHC and IF image captions).
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 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). 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: Secreted protein: tissue RNA and protein sites may differ (HPA tissue IHC); Isoform / epitope: No isoforms; mature chain 24–94 follows signal cleavage (UniProt); Regulation: Stimulated PBMCs only; not resting (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.