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CCR7 / C-C chemokine receptor type 7 · IHC design guide
Design Immunohistochemistry for CCR7
Plan CCR7 paraffin-section IHC around cytoplasmic staining in inflammatory-cell subsets (HPA tissue IHC). Use lymph node non-germinal center cells as a positive reference and adipocytes as a negative reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCR7 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 signal (HPA tissue IHC); membrane receptor (UniProt) | |
| Staining pattern | Cytoplasmic staining in inflammatory-cell subsets (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00390-2) | |
| Positive control | Lung+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Changing inflammatory-cell numbers can alter apparent signal (HPA tissue IHC) | |
| Regulation | EBV infection up-regulates B-cell CCR7 (UniProt) | |
| Isoform / epitope | No isoforms annotated; extracellular vs cytoplasmic epitopes matter (UniProt) |
Section 1
Recommended CCR7 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A00390-2). The four published CCR7 IHC protocols below provide tissue-specific examples (PMC6141431; PMC4008409; PMC8149363; PMC3923395).
| Sample | Paraffin-embedded human lymphoma tissue; fixative not specified (datasheet A00390-2) |
| Fixation | Image fixative and duration unreported (datasheet A00390-2); 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 A00390-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00390-2) |
| Primary antibody | Rabbit anti-CCR7, 2-5 μg/ml (datasheet A00390-2) |
| Primary incubation | Overnight at 4 °C (datasheet A00390-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A00390-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCR7-positive staining in alveolar cells of lung (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in subsets of inflammatory cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A00390-2). Citrate pH 6 is a published alternative for mouse tissue (PMC3923395).
Section 2
What Is the Expected CCR7 Staining Pattern?
CCR7 is a cell membrane receptor with 7 transmembrane segments (UniProt P32248 topology). In tissue IHC, HPA describes cytoplasmic staining in subsets of inflammatory cells, including strong staining in lymphoid tissue cell populations (HPA tissue IHC). HPA rates its tissue IHC evidence Enhanced, with medium consistency between antibody staining and RNA expression; interpret compartment and cell identity together (HPA tissue IHC).
| Strong staining in subsets of non-germinal center cells in lymph node or tonsil, or cells in splenic white pulp (HPA tissue IHC). | These are reported high-staining populations (HPA tissue IHC). A cytoplasmic pattern fits HPA's tissue description; a membrane outline is biologically plausible from CCR7's topology, but the supplied tissue IHC summary does not require a visible outline (HPA tissue IHC; UniProt P32248 topology). |
| Predominantly nuclear staining, or a uniform organellar pattern across the IHC section. | Neither is the reported tissue IHC pattern (HPA tissue IHC). Check the matched positive tissue and negative detection control before assigning such staining to CCR7 (general IHC practice). The mitochondrial ICC-IF report concerns a different application and should be assessed separately (HPA subcellular ICC-IF). |
| Strong signal in respiratory epithelial cells or adipocytes, especially when expected lymphoid populations are unstained. | HPA reports no detection in bronchial respiratory epithelial cells and adipocytes in its listed tissues (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; compare with a no-primary control and the expected positive tissue (general IHC practice). |
| Diffuse color over cells and surrounding tissue without identifiable stained cell subsets. | This cannot establish the selective inflammatory-cell pattern reported by HPA (HPA tissue IHC). Review blocking, primary-antibody concentration, washes, and chromogen development against a no-primary control (general IHC practice). |
| No staining in lymph-node non-germinal center cells or splenic white-pulp cells. | Both are reported high-staining populations, so a blank result warrants a run-level check (HPA tissue IHC). Examine tissue preservation, antigen-retrieval conditions, antibody compatibility, and detection controls as general IHC workflow variables; no CCR7-specific fixation effect is established here (general IHC practice). |
💡Expected CCR7 appearanceCall positive tissue IHC when identifiable cell subsets show cytoplasmic staining, potentially with a membrane outline, strongest in reported lymphoid populations; diffuse tissue-wide or predominantly nuclear color is suspect (HPA tissue IHC; UniProt P32248 topology).
| Compartment and epitope location | CCR7 has 7 membrane-spanning segments and extracellular and cytoplasmic regions (UniProt P32248 topology). An antibody's epitope and section preparation can affect which staining pattern is visible (general IHC practice); no epitope for the intended antibody is supplied. |
| Tissue and cell selection | Non-germinal center cells in lymph node and tonsil, splenic white-pulp cells, and lung alveolar cells are reported High (HPA tissue IHC). Bone-marrow hematopoietic cells are reported Low; a faint result there is a weaker check of the assay (HPA tissue IHC). |
| Evidence strength | The tissue profile has Enhanced reliability but only medium antibody-staining/RNA consistency (HPA tissue IHC). HPA lists CAB003796 as IHC Enhanced (HPA antibodies); that status does not validate a different antibody or resolve every unexpected cell-level result. |
| Processing and modification | UniProt records a 1–24 signal peptide, a 25–378 mature chain, and one glycosylation site at residue 36 (UniProt P32248). These annotations identify structural context; they do not establish a retrieval requirement, shedding pattern, or fixation sensitivity. |
| IF/ICC Q&A: Should mitochondrial fluorescence be called the expected tissue IHC pattern? | No. HPA reports enhanced mitochondrial localization in ICC-IF images from A-549, U2OS, and Rh30, while its tissue IHC profile describes cytoplasmic staining in inflammatory-cell subsets (HPA subcellular ICC-IF; HPA tissue IHC). Assess IF/ICC on its own guide page; the supplied sources do not explain the discrepancy. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive lymph-node or tonsil control is blank. | The run may have lost antigen detection; those non-germinal center cells are reported High (HPA tissue IHC). | Check section quality, retrieval record, primary-antibody identity, and detection reagents with established IHC controls (general IHC practice). Do not infer CCR7-specific fixation failure. |
| Only weak color appears in bone-marrow hematopoietic cells. | That population is reported Low, so it offers limited evidence that the assay can detect a strong positive (HPA tissue IHC). | Compare the same run with lymph-node or tonsil non-germinal center cells reported High (HPA tissue IHC); assess staining strength before changing conditions (general IHC practice). |
| Color appears across the section, including the no-primary control. | Primary-antibody binding cannot explain color in that control; endogenous detection activity or detection-reagent background is possible (general IHC practice). | Check the detection system and its appropriate endogenous-activity block, then repeat the control alongside a reported positive tissue (general IHC practice; HPA tissue IHC). |
| Diffuse haze obscures identifiable cell boundaries, while the no-primary control is clear. | Excess primary reagent, insufficient washing, or prolonged chromogen development can raise nonspecific background (general IHC practice). | Review the antibody's IHC instructions and titrate concentration or development time with paired positive and negative controls (general IHC practice). No dilution is supplied here. |
| Strong nuclear color dominates otherwise plausible lymphoid staining. | A nuclear-dominant pattern does not match HPA's cytoplasmic tissue profile or UniProt's membrane annotation (HPA tissue IHC; UniProt P32248). | Inspect counterstain and chromogen distribution, then compare a no-primary control and a reported positive tissue before scoring CCR7 (general IHC practice; HPA tissue IHC). |
| Many bronchial respiratory epithelial cells stain strongly. | HPA reports those cells as Not detected; cross-reactivity or background needs assessment (HPA tissue IHC; general IHC practice). | Compare cell morphology and no-primary staining, then confirm that expected lymphoid cell populations stain in the same run (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing CCR7 staining
A00390-2 · IHCIHC analysis of CCR7 using anti-CCR7 antibody (A00390-2). CCR7 was detected in a paraffin-embedded section of human lymphoma tissue. Heat me
M00390 · IHCImmunohistochemical analysis of paraffin-embedded human spleen, using CCR7 Antibody.
M00390 · IFImmunofluorescent analysis of Hela cells, using CCR7 Antibody .
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000126353-CCR7/tissue/cerebral+cortex#imid_1297972
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000126353-CCR7/tissue/colon#imid_1297898
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000126353-CCR7/tissue/liver#imid_1297891
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000126353-CCR7/tissue/kidney#imid_1297892
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA031383; green = CCR7, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA031383; green = CCR7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCR7 IHC & IF
🧪Run lymph node first: non-germinal center cells should stain strongly (HPA: High in lymph node non-germinal center cells). Use adipose tissue as the negative comparator, scoring adipocytes rather than any interspersed immune cells (HPA: Not detected in adipocytes; UniProt P32248: lymphocyte expression); on the lymph node slide, use unstained neighboring cells to assess background without assuming a particular cell type is CCR7-negative (standard IHC practice).
Positive control tissue: Lung (Alveolar cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCR7 in A-549, U2OS, Rh30, with annotated localisation: Mitochondria (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and format-matched rabbit isotype controls, plus a matched CCR7-knockout biological control if available (hero caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection and compare any staining in lymph node with the no-primary control (hero caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A00390-2 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU tissue-IHC caption). Lymph node contains mixed cell populations, so score the HPA-designated non-germinal center cells rather than treating all unstained cells as assay failure (HPA: High in lymph node non-germinal center cells).
HPA tissue IHC evidence for CCR7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Lung | Alveolar cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Skin | Cells in corneal layer | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in white pulp | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | 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 → |
| Appendix | Endocrine 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 CCR7 IHC Tips
Troubleshoot CCR7 staining in paraffin sections by checking retrieval, cell identity, staining compartment and controls before comparing signal across samples.
How should I retrieve CCR7 in paraffin sections when staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A00390-2). The selected paraffin section of human lymphoma was stained after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A00390-2). If staining remains weak, compare a carefully controlled alternative retrieval condition on adjacent sections while holding antibody concentration and detection constant; assess tissue damage alongside signal. Include a known positive lymphoid section and a primary antibody omission control, so stronger staining is not mistaken for better specificity (UniProt P32248 tissue specificity; HPA: high in lymph node non-germinal center cells).
Can I infer the best fixative for CCR7 from the lymphoma image?
No target specific fixation sensitivity is established here: the selected lymphoma caption describes a paraffin section but does not state its fixative (datasheet A00390-2). Record the fixative, fixation duration and processing schedule for every specimen, then compare similarly processed sections under the same EDTA pH 8.0 retrieval condition (datasheet A00390-2). When fixation histories differ, run a small matched comparison with a positive lymphoid control before attributing weaker staining to CCR7 abundance (UniProt P32248 tissue specificity). Judge preservation of cell morphology and background alongside staining intensity; neither a positive IHC pattern nor receptor topology establishes fixation tolerance (HPA: inflammatory cell staining; UniProt P32248 topology).
Where should CCR7 staining appear, and how should cytoplasmic signal be judged?
Expect a membrane associated component because CCR7 is annotated as a cell membrane receptor with 7 transmembrane segments (UniProt P32248 localisation and topology). Cytoplasmic staining can also occur in tissue IHC: the HPA profile describes expression in subsets of inflammatory cells (HPA tissue IHC profile). Score membrane and cytoplasmic patterns separately, and compare each with the distribution of the relevant cells on the counterstained section; diffuse colour across unrelated cells warrants a background check. HPA reports mitochondrial localisation in its cell based IF images, so compartment alone cannot settle an apparent conflict between assays (HPA subcellular: mitochondria, enhanced).
How does CCR7 topology affect interpretation of an antibody epitope?
Check the antibody's stated immunogen or epitope before deciding whether surface staining is required; the supplied caption does not identify an epitope (datasheet A00390-2). CCR7 has extracellular and cytoplasmic regions separated by 7 transmembrane segments, a signal peptide at residues 1–24, and an annotated glycosylation site at residue 36 (UniProt P32248 topology and processing). Retrieval may expose these regions differently in paraffin sections, so compare staining patterns under consistent conditions without assigning a particular epitope to this antibody. The record annotates 0 isoforms; that is an annotation count, not proof that every staining difference reflects processing or specificity (UniProt P32248 isoforms).
How can I check CCR7 localisation with a multiplex IF experiment?
Use IF as a complementary check and multiplex CCR7 with a validated marker for the expected lymphoid cell population, since the record reports expression in activated B and T lymphocytes (UniProt P32248 tissue specificity). Choose fluorophores in channels with low measured tissue autofluorescence, and inspect single stained controls before interpreting overlap; retain an unstained section to map intrinsic fluorescence. If the antibody epitope is cytoplasmic, test gentle permeabilisation; for an extracellular epitope, compare with minimal permeabilisation, guided by the antibody's documented epitope rather than an assumed location (UniProt P32248 topology). Interpret membrane and intracellular signals separately because tissue IHC and cell based IF report different predominant compartments (HPA tissue IHC profile; HPA subcellular: mitochondria).
What should I adjust when brown signal obscures CCR7 positive cells?
First inspect a primary antibody omission control for detection related colour, then check endogenous peroxidase blocking and the duration of DAB development; these are general chromogenic IHC controls. The selected lymphoma procedure used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase conjugated secondary antibody for 30 minutes at 37°C (datasheet A00390-2). If background persists, titrate primary antibody and shorten chromogen development while preserving a positive control on the same run. Assess whether apparent widespread colour follows tissue edges, folds or damaged areas rather than the reported subset of inflammatory cells (HPA tissue IHC profile).
How should I quantify CCR7 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since the tissue IHC profile reports CCR7 in subsets of inflammatory cells (HPA tissue IHC profile). Within a prespecified viable area, report the percentage of positive target cells and an intensity based H-score from 0–300, or count positive cells per mm² when cell density is the main outcome. Normalise positive counts to all eligible cells of the same type, and record sampled area, tissue region and exclusion of folds or necrosis. Keep retrieval, antibody concentration, DAB development and score thresholds constant across cases; the selected procedure specifies EDTA pH 8.0 and 2 μg/ml primary antibody (datasheet A00390-2).
How can I distinguish plausible CCR7 staining from artefact?
A plausible result should track an appropriate cell population and show a reproducible pattern across well preserved areas; CCR7 is expressed in lymphoid tissues and activated B and T lymphocytes (UniProt P32248 tissue specificity). Use high staining non-germinal center cells in lymph node or tonsil as contextual positives, while remembering that HPA reports medium consistency between staining and RNA data (HPA tissue IHC positives and reliability). Review unexpected uniform staining of unrelated cells, isolated edge staining, necrotic areas and signal in the primary antibody omission control before calling a case positive. Treat a mitochondrial only pattern cautiously: UniProt annotates cell membrane localisation, whereas HPA cell based IF reports mitochondria (UniProt P32248 localisation; HPA subcellular: mitochondria, enhanced).
Boster reagents
Best CCR7 / C-C chemokine receptor type 7 IHC Antibodies
Anti-CCR7 antibodies have IHC images from human lymphoma and spleen paraffin sections (A00390-2 and M00390 image captions); IF images cover human spleen tissue and HeLa cells (A00390 and M00390 image captions).
Real IHC data
Anti-CCR7 Antibody
Cat # A00390-2
Real IHC data
Anti-CCR7 Rabbit Monoclonal Antibody
Cat # M00390
A00390-2 will render with an IHC image from a paraffin-embedded human lymphoma section (A00390-2 image caption). M00390 will render with an IHC image from paraffin-embedded human spleen (M00390 image caption).
Which to pick: For human tissue IHC, choose A00390-2 when its documented EDTA retrieval and DAB workflow fits your study (A00390-2 image caption); the caption reports a paraffin section but does not report the fixative (A00390-2 image caption). For IF/ICC, choose M00390 because both applications are listed and it has a HeLa IF image (M00390 application list and IF image caption). M00390 is also the cross-species option: it is a monoclonal listed as reactive with human, mouse and rat, although its IHC caption documents only a paraffin-embedded human spleen section and does not report the fixative (M00390 catalog entry and IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P32248 (CCR7_HUMAN, C-C chemokine receptor type 7).
- Human Protein Atlas. CCR7 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCR7 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. CCR7 antibody validation summary (3 antibodies).
- Investigation of CCR7 Marker Expression Using Immunohistochemical Method and Its Association with Clinicopathologic Properties in Patients with Breast Cancer. International journal of hematology-oncology and stem cell research 2018 — PMC6141431.
- CC-Chemokine receptor CCR7: a key molecule for lymph node metastasis in esophageal squamous cell carcinoma. BMC cancer 2014 — PMC4008409.
- Adipose-specific C-C motif chemokine ligand (CCL) 19 overexpression drives the mice to both insulin resistance and weight gain. BMJ open diabetes research & care 2021 — PMC8149363.
- Increased expression of the homeostatic chemokines CCL19 and CCL21 in clinical and experimental Rickettsia conorii infection. BMC infectious diseases 2014 — PMC3923395.
- PubMed PMID:8383238 — UniProt-cited evidence.
- PubMed PMID:7851893 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCR7 (C-C chemokine receptor type 7). Expected localisation Cytoplasmic signal (HPA tissue IHC); membrane receptor (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00390-2). Validated IHC antibody: A00390-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Alveolar cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lymphoma tissue; fixative not specified (datasheet A00390-2); 2. Fixation: Image fixative and duration unreported (datasheet A00390-2); 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 A00390-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00390-2); 8. Primary antibody: Rabbit anti-CCR7, 2-5 μg/ml (datasheet A00390-2); 9. Primary incubation: Overnight at 4 °C (datasheet A00390-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Changing inflammatory-cell numbers can alter apparent signal (HPA tissue IHC); Isoform / epitope: No isoforms annotated; extracellular vs cytoplasmic epitopes matter (UniProt); Regulation: EBV infection up-regulates B-cell CCR7 (UniProt). Evidence: UniProt P32248, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A00390-2, PMC6141431, PMC4008409. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCR7/immunohistochemistry-ihc-guide.