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CCL19 / C-C motif chemokine 19 · IHC design guide
Design Immunohistochemistry for CCL19
Plan chromogenic CCL19 IHC in paraffin sections using the IHC-validated antibody (datasheet A01605-2). Compare staining with HPA tissue controls, and interpret cytoplasmic immune-cell signal in light of CCL19 secretion (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCL19 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 | Immune-cell cytoplasm (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01605-2) | |
| Positive control | Appendix+2 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 | Secreted CCL19 may stain away from its RNA source (HPA tissue IHC) | |
| Regulation | High in lymph nodes, thymus, appendix (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; map epitopes to mature chain 22–98 (UniProt) |
Section 1
Recommended CCL19 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet: A01605-2). The options summarize published CCL19 IHC protocols (PMC3923395; PMC11102069; PMC10683612; PMC4572056).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet A01605-2) |
| Fixation | Image fixative and duration unreported (datasheet A01605-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A01605-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01605-2) |
| Primary antibody | Rabbit anti-CCL19, 2-5 μg/ml (datasheet A01605-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01605-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01605-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCL19-positive staining in germinal center cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in immune cells. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A01605-2). To reproduce a published protocol, use that article’s stated retrieval conditions.
Section 2
What Is the Expected CCL19 Staining Pattern?
CCL19 is a secreted protein with no transmembrane segment (UniProt Q99731). In tissue IHC, expect chiefly cytoplasmic staining in immune cells, including medium staining in appendix and tonsil germinal center cells; small intestine endocrine cells also show medium staining (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while noting medium agreement between antibody staining and RNA expression because secretion can separate protein from its site of synthesis (HPA tissue IHC).
| Cytoplasmic chromogen in appendix or tonsil germinal center cells. | This fits the reported medium staining and immune cell profile (HPA tissue IHC). Judge the cells and compartment together; staining elsewhere does not establish its source in a secreted protein (UniProt Q99731; HPA tissue IHC). |
| Predominantly nuclear staining or a crisp, continuous cell membrane outline. | Neither is the reported tissue pattern (HPA tissue IHC). CCL19 is secreted and lacks a transmembrane segment (UniProt Q99731); review morphology and controls before calling such a pattern specific. |
| Strong staining in cells reported as undetected, such as adipose adipocytes or bronchial respiratory epithelium. | This conflicts with those HPA tissue observations (HPA tissue IHC). Consider cross-reactivity or endogenous chromogenic detection activity, and compare a no-primary control (general IHC practice). |
| Broad, poorly localized color across cells and extracellular spaces. | CCL19 secretion makes extracellular location plausible (UniProt Q99731), but diffuse color alone cannot establish specific staining. Check whether the signal follows tissue structures and whether a no-primary control also colors them (general IHC practice). |
| No staining in appendix or tonsil germinal center cells. | Those cells show medium staining in HPA tissue IHC (HPA tissue IHC). A blank result warrants a check of tissue preservation, retrieval, antibody and detection conditions, and the positive control; it does not alone prove CCL19 absence (general IHC practice). |
💡Expected CCL19 appearanceCall a positive result when identifiable appendix or tonsil germinal center cells show chiefly cytoplasmic staining of roughly medium intensity (HPA tissue IHC); widespread nuclear or uniform membrane staining does not match the reported pattern (HPA tissue IHC; UniProt Q99731).
| Secreted protein and precursor processing (UniProt Q99731). | CCL19 has a signal peptide at residues 1–21 and a mature chain at 22–98, with no transmembrane segment (UniProt Q99731). Assess cytoplasmic tissue staining without assuming all extracellular signal marks the producing cell (HPA tissue IHC; UniProt Q99731). |
| Tissue and cell context (HPA tissue IHC). | Appendix and tonsil germinal center cells and small intestine endocrine cells have medium staining; spleen red pulp and lymph node non-germinal center cells have low staining (HPA tissue IHC). Select and compare controls at the cell level. |
| RNA and protein agreement (HPA tissue IHC; UniProt Q99731). | HPA calls the IHC pattern Enhanced but reports medium consistency with RNA because secreted protein and its RNA can occupy different tissue locations (HPA tissue IHC). UniProt reports high expression in lymph nodes, thymus, and appendix (UniProt Q99731); that does not require equally strong staining in every cell. |
| Antibody validation scope (HPA antibodies). | HPA067758 is listed as rabbit polyclonal with Enhanced IHC validation (HPA antibodies). That status supports the reported tissue pattern; it does not establish specificity for every new specimen, staining condition, or unexpected compartment (HPA tissue IHC; general IHC practice). |
| IF/ICC Q&A: is a cellular fluorescence pattern established? | HPA labels CCL19 secreted but gives no main subcellular location or cell line ICC-IF images, and lists no ICC status for HPA067758 (HPA subcellular; HPA antibodies). Use the tissue IHC pattern as context, not as an IF/ICC validation claim. |
| Situation | Likely cause | Next action |
|---|---|---|
| Appendix or tonsil germinal center cells are blank (HPA tissue IHC). | A failed staining run or unsuitable section can mimic absence (general IHC practice); these cells have medium HPA staining (HPA tissue IHC). | Confirm the positive control and tissue morphology, then review the chosen retrieval, primary antibody, and detection steps against their validated IHC instructions (general IHC practice). |
| Signal is faint but confined to lymph node non-germinal center cells or spleen red pulp (HPA tissue IHC). | HPA reports low staining in those cells (HPA tissue IHC); weak color there alone need not indicate a failed run. | Compare an appendix or tonsil germinal center positive control in the same run before changing detection conditions (HPA tissue IHC; general IHC practice). |
| Diffuse color obscures cellular boundaries. | Background staining may obscure interpretation (general IHC practice); secretion also prevents extracellular location alone from identifying producing cells (UniProt Q99731). | Inspect the no-primary control and tissue morphology; review blocking, washing, and detection conditions if control color is present (general IHC practice). |
| Adipose adipocytes or bronchial respiratory epithelial cells stain strongly. | Both are reported as undetected in HPA tissue IHC (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Check a no-primary control, assess whether the color follows the reported immune cell pattern, and verify the unexpected result independently before scoring it as CCL19 (HPA tissue IHC; general IHC practice). |
| Nuclear or continuous membrane staining dominates. | It conflicts with the reported cytoplasmic immune cell profile (HPA tissue IHC) and with the absence of a transmembrane segment (UniProt Q99731). | Recheck cell identification and no-primary controls; treat the compartment as unconfirmed until it is independently supported (general IHC practice). |
| Lymph node staining appears lower than expected from its expression description. | UniProt reports high lymph node expression, while HPA reports low staining in lymph node non-germinal center cells and cautions that secreted protein may differ in location from RNA (UniProt Q99731; HPA tissue IHC). | Compare the same cell population across sections and a positive tissue control; report the observed cells and intensity instead of inferring protein distribution from tissue-wide expression (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CCL19 staining
A01605-2 · IHCIHC analysis of MIP-3 Beta/CCL19 using anti-MIP-3 Beta/CCL19 antibody (A01605-2). MIP-3 Beta/CCL19 was detected in a paraffin-embedded secti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172724-CCL19/tissue/cerebral+cortex#imid_20025681
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172724-CCL19/tissue/colon#imid_20025599
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172724-CCL19/tissue/liver#imid_20025600
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172724-CCL19/tissue/kidney#imid_20025609
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCL19 IHC & IF
🧪Run appendix first and assess germinal center cells for staining (HPA: Medium in appendix germinal center cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the appendix slide, compare nearby cells outside germinal centers for background staining, without treating them as confirmed CCL19-negative cells (HPA: germinal center cells are the annotated positive population).
Positive control tissue: Appendix (Germinal center 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 CCL19; derive a cell-line control from the positive tissue's cell type (Germinal center cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-matched rabbit IgG isotype controls, plus a CCL19-knockout specimen if available (caption: rabbit primary antibody; standard IHC practice). Check endogenous peroxidase activity before interpreting DAB staining in appendix tissue (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected IHC caption does not state the fixative (selected-SKU tissue-IHC caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 for a paraffin spleen section; whether that retrieval is required in appendix or whether frozen-section IF is easier is unreported (selected-SKU tissue-IHC caption). Interpret diffuse or extracellular staining cautiously because CCL19 is secreted, and check for tissue background with the controls (UniProt Q99731: Secreted; standard IHC practice).
HPA tissue IHC evidence for CCL19
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Secreted protein, tissue location of RNA and protein is expected to differ.). 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 | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Small intestine | Endocrine cells | Medium | Protein (IHC) | HPA → |
| Tonsil | Germinal center 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCL19 IHC Tips
Troubleshoot CCL19 staining in paraffin sections by checking retrieval, controls, cellular context and detection before interpreting signal intensity.
Which retrieval conditions should I use when CCL19 staining is weak?
Use heat-mediated retrieval in EDTA at pH 8.0 for paraffin-section CCL19 IHC (datasheet A01605-2). The selected image used this retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so preserve those documented conditions while troubleshooting (caption A01605-2). If staining remains weak, compare retrieval duration or a second buffer on adjacent sections, changing one variable at a time (standard IHC practice). Include a known positive section in each run and watch for tissue damage or diffuse background as retrieval increases (standard IHC practice).
How should I troubleshoot CCL19 staining across differently fixed specimens?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the selected image describes a paraffin-embedded spleen section but does not state its fixative (caption A01605-2). Record each specimen’s fixative and fixation interval, then compare sections processed and stained in the same run (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary incubation at 2 μg/ml overnight at 4°C for the initial comparison (caption A01605-2). If staining varies, assess tissue preservation and staining controls before attributing the difference to CCL19 abundance or a target-specific fixation effect (standard IHC practice).
Where should I expect CCL19 signal in a paraffin section?
Interpret cellular staining alongside CCL19’s secreted status and absence of a transmembrane segment (UniProt Q99731 topology and subcellular annotation). The tissue atlas describes cytoplasmic expression in immune cells, while warning that protein and RNA locations can differ for a secreted protein (HPA tissue IHC). Its reported staining includes medium signal in appendix and tonsil germinal center cells, and low signal in spleen red-pulp cells (HPA tissue IHC). Examine cell morphology and neighboring structures at higher magnification; diffuse extracellular color alone cannot identify the producing cell, so compare cellular and surrounding signal with matched controls (standard IHC practice; UniProt Q99731 secreted annotation).
Could processing or isoforms explain discordant CCL19 staining?
CCL19 is annotated as a 98-amino-acid precursor with a signal peptide at residues 1–21 and a mature chain at 22–98 (UniProt Q99731 processing). The supplied record lists no isoforms, glycosylation sites or modified residues, but it does not specify the catalog antibody’s epitope (UniProt Q99731; supplied antibody evidence). Ask whether the immunogen or mapped epitope recognizes the mature chain before treating a negative section as evidence of absent protein (standard IHC interpretation). Compare another independently validated epitope or an appropriate control section if available, and keep retrieval and detection conditions matched across comparisons (standard IHC practice).
How can I check CCL19 localisation by multiplex immunofluorescence?
Pair CCL19 with a validated immune-cell marker to examine the cell populations implicated by the tissue atlas, while recognizing that secreted protein may appear beyond its source cell (HPA tissue IHC; UniProt Q99731 secreted annotation). Choose a fluorophore whose emission is distinguishable from the section’s autofluorescence, and image single-stain and unstained controls before assessing colocalisation (standard IF practice). For an intracellular epitope, test gentle permeabilisation; an extracellular accessible epitope may be assessed without it, but the catalog antibody’s epitope is unspecified (standard IF practice; supplied antibody evidence). This is an IF troubleshooting comparison, not an IF validation claim for the catalog antibody (supplied antibody evidence).
What should I change when CCL19 DAB staining is diffuse?
The selected paraffin-section image used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, a peroxidase-conjugated secondary for 30 minutes at 37°C, and DAB development (caption A01605-2). Add a no-primary control and an endogenous-peroxidase block to assess nonspecific secondary signal and tissue enzyme activity (standard chromogenic IHC practice). If background persists, titrate the primary and shorten chromogen development while keeping a positive control in the same run (standard IHC practice). Because CCL19 is secreted, diffuse staining cannot by itself establish which cells produced it; assess cellular detail and control staining before assigning localisation (UniProt Q99731 subcellular annotation; standard IHC interpretation).
How should I quantify CCL19 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and cell classes before scoring, then use percent positive cells, an H-score, or positive-cell density per mm² according to the question (standard IHC quantification practice). Normalize cell counts to the number of evaluable cells or measured tissue area, and compare like regions under matched staining and imaging conditions (standard IHC quantification practice). Report cellular signal separately from extracellular DAB because CCL19 is secreted and the two measurements answer different questions (UniProt Q99731 subcellular annotation; standard IHC interpretation). Record background thresholds from controls and exclude folds, damaged edges and necrotic areas consistently across sections (standard IHC quantification practice).
How can I distinguish convincing CCL19 staining from artefact?
Look for reproducible cellular staining in a plausible compartment and compare it with the atlas’s cytoplasmic immune-cell pattern; CCL19 is secreted, so extracellular signal requires cautious attribution (HPA tissue IHC; UniProt Q99731 subcellular annotation). Appendix or tonsil germinal center cells provide reported medium-staining examples, whereas spleen red-pulp cells are reported at low staining (HPA tissue IHC). Reject edge-restricted color, necrotic deposits and signal reproduced in a no-primary control as evidence of specific staining (standard IHC interpretation). Check endogenous peroxidase activity when DAB appears in unexpected cells, and use morphology plus matched controls before calling a biological difference (standard chromogenic IHC practice).
Boster reagents
Best CCL19 / C-C motif chemokine 19 IHC Antibodies
The catalog has one human-reactive anti-CCL19 antibody with real IHC data from a paraffin-embedded human spleen section (A01605-2 image caption; catalog reactivity).
Real IHC data
Anti-MIP-3 Beta/CCL19 Antibody
Cat # A01605-2
A01605-2 is listed for human IHC, flow cytometry and ELISA (catalog applications and reactivity). Its IHC figure shows a paraffin-embedded human spleen section (A01605-2 image caption).
Which to pick: For tissue IHC, choose A01605-2, a rabbit polyclonal antibody listed at 2–5 μg/ml for paraffin sections (catalog host, clonality and IHC dilution). Its spleen IHC caption reports EDTA retrieval at pH 8.0 and a 2 μg/ml primary antibody concentration; the fixative is unreported (A01605-2 image caption). There is no listed IF/ICC or cross-species option: A01605-2 lists human reactivity and IHC, flow cytometry and ELISA, with no IF dilution or figure (catalog applications, reactivity and IF fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q99731 (CCL19_HUMAN, C-C motif chemokine 19).
- Human Protein Atlas. CCL19 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCL19 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. CCL19 antibody validation summary (1 antibodies).
- Increased expression of the homeostatic chemokines CCL19 and CCL21 in clinical and experimental Rickettsia conorii infection. BMC infectious diseases 2014 — PMC3923395.
- Chemokine CCL19 and Its Receptors CCR7 and CCRL1 in Chronic Rhinosinusitis. Journal of inflammation research 2024 — PMC11102069.
- CCL19: a novel prognostic chemokine modulates the tumor immune microenvironment and outcomes of cancers. Aging 2023 — PMC10683612.
- Central Nervous System and Peripheral Expression of CCL19, CCL21 and Their Receptor CCR7 in Experimental Model of Multiple Sclerosis. Archivum immunologiae et therapiae experimentalis 2015 — PMC4572056.
- PubMed PMID:9013939 — UniProt-cited evidence.
- PubMed PMID:9153236 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCL19 (C-C motif chemokine 19). Expected localisation Immune-cell cytoplasm (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01605-2). Validated IHC antibody: A01605-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human spleen tissue; fixative not specified (datasheet A01605-2); 2. Fixation: Image fixative and duration unreported (datasheet A01605-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A01605-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01605-2); 8. Primary antibody: Rabbit anti-CCL19, 2-5 μg/ml (datasheet A01605-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01605-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secreted CCL19 may stain away from its RNA source (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; map epitopes to mature chain 22–98 (UniProt); Regulation: High in lymph nodes, thymus, appendix (UniProt). Evidence: UniProt Q99731, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A01605-2, PMC3923395, PMC11102069. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCL19/immunohistochemistry-ihc-guide.