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CCL19 / C-C motif chemokine 19 · IHC design guide
Design Immunohistochemistry for CCL19
Plan chromogenic CCL19 IHC with the catalog antibody at 2–5 μg/ml (datasheet A01605-2). Assess cytoplasmic staining in immune cells, using appendix or tonsil germinal center cells as positive tissue references (HPA tissue IHC).
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 | Cytoplasmic staining in immune cells (HPA tissue IHC) | |
| Staining pattern | Immune cells show cytoplasmic staining (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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Secreted protein may appear away from its RNA source (HPA tissue IHC) | |
| Regulation | No expression regulator reported (UniProt) | |
| Isoform / epitope | No isoforms; signal 1–21 precedes mature chain 22–98 (UniProt) |
Section 1
Recommended CCL19 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet: A01605-2). The published options below report CCL19 staining in breast cancer tissues and tissue sections (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 heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A01605-2). Citrate pH 6 is a published alternative (PMC4572056).
Section 2
What Is the Expected CCL19 Staining Pattern?
CCL19 is secreted and has no transmembrane segment (UniProt Q99731: subcellular location and topology). In tissue IHC, expect cytoplasmic staining in immune cells, including medium staining of germinal center cells in appendix and tonsil (HPA: tissue IHC). HPA rates tissue staining reliability Enhanced, while noting medium consistency with RNA data because a secreted protein’s RNA and protein locations can differ (HPA: tissue IHC).
| Cytoplasmic staining in appendix or tonsil germinal center cells. | This matches the reported Medium staining in those cells (HPA: tissue IHC). Assess cell identity and compartment together; staining elsewhere in a section does not, by itself, identify the cells that produced a secreted protein (UniProt Q99731: secreted; HPA: tissue IHC reliability description). |
| Predominantly nuclear or sharply membrane-restricted staining. | Treat this as a compartment mismatch: HPA reports cytoplasmic expression in immune cells, and UniProt reports secretion without a transmembrane segment (HPA: tissue IHC; UniProt Q99731: topology). Review morphology and detection controls before interpreting the signal as CCL19. |
| Strong staining in adipocytes or bronchial respiratory epithelium. | These cell populations are reported as Not detected (HPA: tissue IHC). Consider nonspecific binding or endogenous chromogen-producing activity, and check a detection control. Do not treat every nonimmune cell as unexpected: small-intestinal endocrine cells show Medium staining (HPA: tissue IHC). |
| Diffuse color across cells, stroma, and the section edge. | Broad color that obscures cellular boundaries limits scoring. Because CCL19 is secreted, extracellular signal alone cannot establish the producing cell type (UniProt Q99731: secreted). Compare a control lacking primary antibody and inspect whether precipitate, nonspecific binding, or endogenous detection activity explains the distribution. |
| No staining in appendix or tonsil germinal center cells. | This conflicts with their reported Medium staining and makes a negative study section difficult to interpret (HPA: tissue IHC). Check section preservation, retrieval and detection with a concurrently processed control; a low-staining lymph node population is a less decisive reference (HPA: lymph node non-germinal center cells, Low). |
💡Expected CCL19 appearanceCall the result positive when identifiable immune-cell cytoplasm stains in the expected distribution, especially Medium germinal center staining in appendix or tonsil; isolated nuclear, membrane-restricted, or uniformly diffuse color warrants control review (HPA: tissue IHC; UniProt Q99731: topology).
| Secretion and processing | CCL19 has a signal sequence at residues 1–21 and a mature chain at 22–98 (UniProt Q99731: processing). A secreted signal may extend beyond its source cells; position alone cannot identify a producer (UniProt Q99731: secreted). |
| Reference tissue and cell population | Appendix and tonsil germinal center cells are Medium; lymph node non-germinal center cells and splenic red-pulp cells are Low (HPA: tissue IHC). Choose the cell population, rather than the organ name alone, when judging a control. |
| Strength of tissue evidence | HPA assigns Enhanced IHC reliability but reports only medium consistency between staining and RNA expression (HPA: tissue IHC). Its stated reason is that protein and RNA locations can differ for secreted CCL19 (HPA: tissue IHC reliability description). |
| IF/ICC Q: Is a cellular pattern established? | No main subcellular location or ICC-IF image is available (HPA: subcellular). UniProt lists CCL19 as secreted (UniProt Q99731: subcellular location); the tissue IHC pattern should not be presented as an independently observed ICC-IF pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank. | A failed stain run or section-level problem is possible; appendix and tonsil germinal center cells are reported as Medium (HPA: tissue IHC). | Run a known-positive section alongside the sample. If both are blank, check retrieval, primary-antibody dilution and detection as general IHC workflow checks; no CCL19-specific retrieval condition is established here. |
| Positive tissue stains only faintly. | The chosen reference population may naturally be weak: lymph node non-germinal center cells and splenic red-pulp cells are Low (HPA: tissue IHC). | Confirm which cells were scored, then compare with appendix or tonsil germinal center cells, reported as Medium (HPA: tissue IHC), in the same staining run. |
| Color persists when primary antibody is omitted. | The signal is independent of primary-antibody binding and may reflect endogenous detection activity or background from the detection reagents. | Review the detection control and apply blocking appropriate to the chromogenic system. Reassess the tissue only after the control is acceptably clear; this is a general IHC control check. |
| Nuclear or crisp membrane staining dominates. | That pattern conflicts with reported cytoplasmic immune-cell staining and a protein with no transmembrane segment (HPA: tissue IHC; UniProt Q99731: topology). | Check morphology, counterstain and the primary-omission control. Re-evaluate antibody dilution or detection conditions before assigning a CCL19-positive score. |
| Adipocytes or bronchial epithelium stain strongly. | Both are reported as Not detected in the specified cells (HPA: tissue IHC); cross-reactivity or detection background is possible. | Compare with the primary-omission control and an expected positive population. Score unexpected staining separately until its specificity is resolved. |
| Diffuse extracellular color makes cell scoring uncertain. | CCL19 is secreted, so location outside a cell does not identify its producer (UniProt Q99731: secreted); excess chromogen or nonspecific background can also obscure boundaries. | Inspect the control lacking primary antibody, shorten chromogen development if the whole section is dark, and base cellular scoring on identifiable cells with a clear cytoplasmic pattern (HPA: tissue IHC). |
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 look for staining in germinal center cells (HPA: Medium); use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected). On the appendix slide, cells outside the germinal centers should provide a low-background comparison, but do not treat them as confirmed CCL19-negative cells without validation (HPA: only germinal center cells are listed as positive).
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 a no-primary, secondary-only control, host- and class-matched nonimmune rabbit IgG, and CCL19 knockout tissue if available or a validated peptide competition control (selected caption: rabbit primary antibody). Quench endogenous peroxidase before DAB development in appendix sections; block endogenous biotin if using avidin–biotin detection (selected caption: peroxidase detection with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01605-2 paraffin-section caption does not state the fixative. Heat retrieval with EDTA at pH 8.0 is documented for the selected IHC example, but its necessity has not been established (selected caption: heat-mediated EDTA retrieval). Frozen sections and IF/ICC have no matched protocol in the supplied evidence, so neither can be called easier; in appendix, check for endogenous peroxidase background, and interpret diffuse staining cautiously because CCL19 is secreted (UniProt: 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
Use the selected paraffin IHC conditions as a starting point, then assess staining against CCL19 secretion and the documented tissue pattern (caption A01605-2; UniProt Q99731; HPA tissue IHC).
How should I troubleshoot weak CCL19 staining after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections stained with A01605-2 (datasheet A01605-2). The selected spleen image used this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (caption A01605-2). Compare matched sections with and without retrieval to separate insufficient epitope exposure from nonspecific DAB deposition (standard IHC practice). If staining remains weak, adjust heating duration in a small matched series while keeping EDTA at pH 8.0 and the detection workflow constant (standard IHC practice). Evaluate immune-cell cytoplasm and surrounding tissue separately because CCL19 is secreted and lacks a transmembrane segment (UniProt Q99731; HPA tissue IHC).
Could fixation explain inconsistent CCL19 staining between paraffin-section batches?
No CCL19-specific fixation sensitivity is established; the selected caption calls its specimen paraffin embedded but does not report the fixative (caption A01605-2). For new specimens, record fixative and fixation duration, then keep these identical across comparison groups (standard IHC practice). If intensity changes across batches, process adjacent sections together with the same EDTA pH 8.0 retrieval and 2 μg/ml primary concentration (datasheet A01605-2; caption A01605-2). Include a same-run positive tissue control and a no-primary control to distinguish processing loss from detection background (standard IHC practice). Do not interpret a batch difference as altered CCL19 expression until fixation and processing differences are excluded (standard IHC practice).
Which CCL19 staining compartments should I assess in tissue sections?
CCL19 is secreted, has no transmembrane segment, and has a mature 22–98 chain (UniProt Q99731). In paraffin IHC, assess cytoplasmic immune-cell staining as the documented pattern, while evaluating extracellular DAB separately (HPA tissue IHC; UniProt Q99731). Appendix and tonsil germinal-center cells show medium staining, whereas spleen red-pulp cells and lymph-node nongerminal-center cells are low (HPA tissue IHC). Check morphology before calling diffuse extracellular signal positive, since secreted antigen may occur away from its producing cells (UniProt Q99731; standard IHC practice). Nuclear-only staining conflicts with the supplied localization evidence and warrants a no-primary control and retrieval review (UniProt Q99731; HPA tissue IHC; standard IHC practice).
Could CCL19 processing or isoforms explain different IHC patterns?
The annotated precursor is 98 residues, with signal peptide 1–21 and mature chain 22–98 (UniProt Q99731). No alternative isoforms, glycosylation sites or modified residues are annotated, so these records do not explain discrepant staining (UniProt Q99731). The antibody epitope is not given, and the paraffin-tissue image cannot establish whether it recognizes precursor or mature CCL19 (caption A01605-2; UniProt Q99731). Keep EDTA at pH 8.0 and primary antibody at 2 μg/ml when comparing tissues so processing changes do not masquerade as epitope differences (datasheet A01605-2; caption A01605-2; standard IHC practice). Interpret staining changes against matched tissue controls and the documented immune-cell pattern (HPA tissue IHC; standard IHC practice).
How can I assess CCL19 by multiplex IF alongside paraffin IHC?
For IF follow-up, pair CCL19 with CD45 to locate immune cells and inspect morphology (HPA tissue IHC; standard IF practice). Place the weaker signal in a red or far-red channel after checking autofluorescence in an unstained section (standard IF practice). For intracellular CCL19, use mild permeabilisation; for an extracellular epitope, omit permeabilisation on a matched section (UniProt Q99731 secretion and topology; standard IF practice). The selected caption documents paraffin IHC only; titrate the IF primary and include single-stain and no-primary controls (caption A01605-2; standard IF practice). Compare cytoplasmic and extracellular fluorescence separately, since a secreted chemokine need not remain in its producing cell (UniProt Q99731; HPA tissue IHC).
How do I identify and reduce diffuse brown background in CCL19 IHC?
Start with the documented 10% goat-serum block and 2 μg/ml primary antibody, then compare matched no-primary sections (caption A01605-2; standard IHC practice). The selected image used a peroxidase-conjugated secondary for 30 minutes at 37°C and DAB development (caption A01605-2). Add a peroxidase block before HRP detection, and check whether endogenous enzyme activity persists in the no-primary section (standard IHC practice). If diffuse brown signal remains, titrate the primary and shorten DAB development while maintaining the same retrieval and counterstain across sections (standard IHC practice). Judge background against viable tissue away from folds, edges and necrosis; CCL19 secretion can complicate extracellular staining calls (UniProt Q99731; standard IHC practice).
How should I quantify CCL19 IHC across tissue compartments? ⚠ ANSWER MARKED FOR VERIFICATION
Score viable immune-cell cytoplasm using an H-score from 0–300, recording the percentage at each intensity and multiplying by its 0–3 grade (HPA tissue IHC; standard IHC practice). Also report positive cells per mm² of viable tissue and the fraction of immune cells positive (standard IHC practice). Segment germinal centers and red pulp where applicable, because the observations refer to specific compartments (HPA tissue IHC). Keep section thickness, EDTA pH 8.0 retrieval, imaging thresholds and DAB development consistent (datasheet A01605-2; standard IHC practice). Normalize to viable tissue area or counted immune cells, and score extracellular deposition separately from cellular cytoplasm (UniProt Q99731; HPA tissue IHC; standard IHC practice).
When is CCL19 DAB staining convincing rather than artefactual?
Call a positive result when reproducible cytoplasmic staining aligns with immune-cell morphology and the documented tissue pattern (HPA tissue IHC; standard IHC practice). Medium germinal-center staining in appendix or tonsil and low red-pulp staining in spleen provide context, not universal thresholds (HPA tissue IHC). CCL19 is secreted and lacks a transmembrane segment, so crisp membrane or nuclear-only DAB requires control review (UniProt Q99731; standard IHC practice). Exclude edge staining, folds and necrotic debris by comparing intact interior tissue and a no-primary section (standard IHC practice). Investigate widespread brown signal with a peroxidase-block check, and do not equate extracellular deposition with the producing cell (UniProt Q99731; standard IHC practice).
Boster reagents
Best CCL19 / C-C motif chemokine 19 IHC Antibodies
The catalog contains one human-reactive anti-CCL19 antibody with a real IHC image from a paraffin-embedded human spleen section (catalog applications; image caption).
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 image shows a paraffin-embedded human spleen section stained with 2 μg/ml primary antibody (image caption).
Which to pick: Choose A01605-2 for human paraffin-section IHC; it is a rabbit polyclonal antibody with a listed IHC range of 2–5 μg/ml (catalog; image caption). The caption reports heat retrieval in EDTA at pH 8.0, but does not report the fixative (image caption). No SKU in the supplied catalog is validated for IF/ICC or for another species (catalog applications and reactivity).
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 Cytoplasmic staining in immune cells (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 paraffin sections. (standard IHC practice; not target-specific); Caveat: Secreted protein may appear away from its RNA source (HPA tissue IHC); Isoform / epitope: No isoforms; signal 1–21 precedes mature chain 22–98 (UniProt); Regulation: No expression regulator reported (UniProt). Evidence: UniProt Q99731, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A01605-2, PMC10683612, PMC4572056. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCL19/immunohistochemistry-ihc-guide.