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CD209 / CD209 antigen · IHC design guide
Design Immunohistochemistry for CD209
Plan CD209 IHC-P around the selective tissue staining reported by HPA and the cell-membrane localization annotated by UniProt (HPA tissue IHC; UniProt). Start the IHC-validated catalog antibody M01025-1 at 5–10 μg/mL (datasheet) and assess staining by annotated cell type (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CD209 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 | Cell membrane expected; tissue compartment unverified (UniProt; HPA tissue IHC) | |
| Staining pattern | Selective monocyte-derived cell subsets; compartment unreported (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Placenta+3 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 | Placental cytotrophoblast staining complicates DC attribution (HPA tissue IHC) | |
| Regulation | Cell-type-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | 12 isoforms; check cytoplasmic vs extracellular epitope (UniProt) |
Section 1
Recommended CD209 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with two published CD209 protocols in human cardiac tissue (PMC3614732; PMC6159861).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet M01025-1); 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 | Mouse monoclonal (clone 5D7) anti-CD209, 5 - 10 μg/mL (datasheet M01025-1) |
| 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 | CD209-positive staining in cytotrophoblasts of placenta (HPA tissue IHC: High). HPA tissue profile: Selective expression in subsets of monocyte-derived cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval rule); neither article specifies retrieval conditions.
Section 2
What Is the Expected CD209 Staining Pattern?
CD209 is a cell-surface receptor with an extracellular region spanning residues 59–404; UniProt also lists membrane-raft and secreted localisation (UniProt Q9NNX6 topology; subcellular location). In paraffin-section IHC, expect selective staining rather than uniform tissue-wide signal: HPA reports High staining in placental cytotrophoblasts and Medium staining in lung macrophages and bone-marrow hematopoietic cells (HPA tissue IHC; reliability: Supported).
| Distinct staining outlines a subset of cells, with little signal in adjacent cells. | A membrane-associated pattern fits CD209 topology; selective distribution fits the HPA tissue profile (UniProt Q9NNX6 topology; HPA tissue IHC). Compare the stained cell population with tissue morphology before calling the result positive. |
| Strong nuclear staining dominates cells expected to express CD209. | Nuclear localisation is unsupported by the supplied subcellular records (UniProt Q9NNX6 subcellular location; HPA subcellular summary: Membrane). Treat nuclear-only signal as suspect and check controls, detection reagents and slide interpretation (general IHC practice). |
| Adipocytes or bronchial respiratory epithelial cells stain as strongly as expected positive cells. | HPA reports these cells as Not detected (HPA tissue IHC: adipocytes; bronchial respiratory epithelial cells). Consider nonspecific antibody binding or endogenous detection activity; confirm with a no-primary control and a positive tissue control (general IHC practice). |
| Colour spreads evenly across cells, stroma and empty areas. | A field-wide deposit does not match HPA's selective cellular profile (HPA tissue IHC). Review blocking, washes, detection chemistry and counterstain, then compare with a no-primary control (general IHC practice). |
| A properly identified positive tissue shows no convincing cellular signal. | First check that the expected cell population is present: HPA reports High placental cytotrophoblast staining and Medium lung-macrophage staining (HPA tissue IHC). Then review antibody dilution, retrieval and detection controls; no CD209-specific retrieval requirement is supplied (general IHC practice). |
💡Expected CD209 appearanceCall a result positive when staining is selective and predominantly membrane-associated in the expected cells, with HPA reporting High placental cytotrophoblast and Medium lung-macrophage staining; uniform background or nuclear-only staining is suspect (HPA tissue IHC; UniProt Q9NNX6 topology; HPA subcellular summary).
| Epitope location and accessibility | CD209 has a 37-residue cytoplasmic region, a transmembrane segment at 38–58, and an extracellular region at 59–404 (UniProt Q9NNX6 topology). Antibody epitope information is absent, so the detected domain and any retrieval preference cannot be assigned. |
| Cell selection and tissue choice | HPA lists High placental cytotrophoblast staining and Medium bone-marrow hematopoietic, lung-macrophage and testicular late-spermatid staining (HPA tissue IHC). Its profile describes selective expression in subsets of monocyte-derived cells; apply the listed cell-level observations when choosing controls (HPA tissue IHC). |
| IHC antibody evidence | Both listed antibodies, CAB032436 and CAB033831, have IHC status Supported (HPA antibodies). The supplied record does not label either IHC result Enhanced; Supported should not be presented as independent-antibody reproduction (HPA antibodies; HPA validation definitions). |
| Isoforms and glycosylation | UniProt lists 12 isoforms and one glycosylation site at residue 80 (UniProt Q9NNX6). Without an antibody epitope map or isoform-specific staining data, these features cannot explain a particular weak, absent or unexpected IHC signal. |
| IF/ICC interpretation? | HPA summarises CD209 subcellular localisation as Membrane but supplies no ICC-IF image cell lines or main-location assignment (HPA subcellular record). Use membrane localisation as context for IF/ICC interpretation; the supplied record supports no IF/ICC-specific protocol choice. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a placenta section expected to be positive. | The sampled area may lack identifiable cytotrophoblasts, or the staining run may have failed; HPA reports High staining specifically in placental cytotrophoblasts (HPA tissue IHC). | Confirm the cell population by morphology, then check positive-run controls, antibody dilution, retrieval and detection steps (general IHC practice). Do not infer CD209-specific fixation sensitivity from this result. |
| Weak or patchy signal in lung. | HPA reports Medium staining in macrophages, so the expected signal concerns a cell subset rather than every lung cell (HPA tissue IHC). | Locate macrophages morphologically and assess signal within them. Compare a positive tissue and no-primary control before changing conditions (general IHC practice). |
| Prominent colour in adipocytes or bronchial epithelium. | Those cell types are listed as Not detected (HPA tissue IHC); nonspecific binding or endogenous detection activity may contribute (general IHC practice). | Run a no-primary control, check blocking and inspect detection chemistry before interpreting the deposit as CD209 (general IHC practice). |
| Apparent staining is mainly nuclear. | The supplied localisation records support membrane-associated CD209, while its annotated transmembrane segment spans residues 38–58 (HPA subcellular summary; UniProt Q9NNX6 topology). | Treat nuclear-only staining cautiously; compare the no-primary control and inspect whether membrane-associated signal remains in expected cells (general IHC practice). |
| Diffuse brown haze obscures cellular outlines. | Broad haze conflicts with HPA's selective tissue profile and may reflect background from blocking, washing or detection (HPA tissue IHC; general IHC practice). | Compare a no-primary control, review reagent exposure and washes, and score only discernible cellular staining (general IHC practice). |
| Two antibodies give different cell patterns. | Both listed antibodies are IHC Supported, but the supplied evidence does not establish their epitope equivalence or an Enhanced match (HPA antibodies; HPA validation definitions). | Compare each pattern against HPA's named positive and Not detected cell types; review controls and antibody documentation before assigning either pattern to CD209 (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CD209 staining
M01025-1 · IHCImmunohistochemistry of DC-SIGN in human lymph node tissue with DC-SIGN antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000090659-CD209/tissue/cerebral+cortex#imid_8098964
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000090659-CD209/tissue/colon#imid_8085866
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000090659-CD209/tissue/liver#imid_8085867
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000090659-CD209/tissue/kidney#imid_8085852
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CD209 IHC & IF
🧪Run placenta first; cytotrophoblasts should stain (HPA: High in placental cytotrophoblasts). Use adipose tissue as the negative tissue because adipocytes are not detected (HPA: Not detected in adipocytes); other cells on the placenta slide should remain at background unless their CD209 expression is independently established.
Positive control tissue: Placenta (Cytotrophoblasts, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CD209; derive a cell-line control from the positive tissue's cell type (Cytotrophoblasts) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control, an isotype control matched to the primary antibody’s host and immunoglobulin class, and CD209-knockout material as a biological negative where available (standard IHC practice). For placental chromogenic IHC, quench endogenous peroxidase and check for blood-associated background (standard IHC practice).
⚠️Feasibility: The supplied evidence reports no target-specific fixation window or fixation effect, and the selected M01025-1 tissue-IHC caption does not state a fixative (caption: fixative unreported). Retrieval dependency is unreported, so the evidence does not establish whether frozen sections or IF would be easier. Placental blood can cause chromogenic background; evaluate it with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for CD209
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — External characterization data supports antibody staining. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | 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 → |
| Appendix | Glandular 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 CD209 IHC Tips
Troubleshoot CD209 staining in paraffin sections by checking retrieval, cell identity, membrane localisation and controls before interpreting signal (UniProt Q9NNX6; HPA tissue IHC).
How should I retrieve CD209 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page antigen retrieval setting). Let sections cool in the retrieval buffer, then wash consistently so temperature differences do not complicate comparisons (standard IHC practice). CD209 has an extracellular region spanning residues 59–404 and a membrane segment at 38–58, but topology alone does not establish which retrieval condition preserves this antibody’s epitope (UniProt Q9NNX6 topology). If staining remains weak, compare a carefully controlled alternative retrieval condition on adjacent sections while keeping detection and exposure to chromogen constant (standard IHC practice). Score signal in expected cells alongside background, since stronger overall staining can reflect nonspecific retrieval effects (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or uneven CD209 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence, so do not assign weak CD209 staining to a particular fixative or fixation duration (supplied evidence). The lymph node image for M01025-1 reports staining at 5 µg/mL but does not state its fixative (M01025-1 tissue-IHC caption). For paraffin IHC, compare sections with documented, consistent fixation and processing histories before changing antibody concentration or detection chemistry (standard IHC practice). Include an expected positive tissue and a reagent control in the same run, then inspect whether uneven staining follows section thickness, folds or edges (HPA tissue IHC; standard IHC practice). Record processing differences as possible explanations rather than evidence of a CD209-specific fixation effect (supplied evidence; standard IHC practice).
What staining pattern should count as plausible CD209 localisation?
Prioritise cell-associated membrane staining because CD209 is annotated at the cell membrane and in membrane rafts (UniProt Q9NNX6 subcellular; HPA subcellular). Its cytoplasmic tail spans residues 1–37, its transmembrane segment 38–58, and its extracellular region 59–404 (UniProt Q9NNX6 topology). Some intracellular puncta can be biologically plausible because CD209 mediates endocytosis and returns to the cell surface, but puncta alone need cellular and control context (UniProt Q9NNX6 function; standard IHC practice). Compare staining with cell morphology and adjacent negative areas; broad nuclear staining is inconsistent with the recorded membrane localisation (UniProt Q9NNX6 subcellular; standard IHC practice). Record surface and intracellular patterns separately when scoring, rather than merging them into one intensity value (standard IHC practice).
How can isoforms and epitope position affect CD209 IHC?
CD209 has 12 annotated isoforms, so confirm which sequence region the catalog antibody recognises before assuming it detects every form (UniProt Q9NNX6 isoforms; standard IHC practice). The extracellular region spans residues 59–404, including a C-type lectin domain at 263–378, while the cytoplasmic tail spans 1–37 (UniProt Q9NNX6 topology and domains). A glycosylation site is annotated at residue 80, but the supplied evidence does not establish whether it alters this antibody’s staining (UniProt Q9NNX6 glycosylation; supplied evidence). If epitope information is available, map it to the isoform sequences and record whether retrieval changes the expected cellular pattern (standard IHC practice). Do not interpret discordant staining as isoform-specific without epitope and control evidence (standard IHC practice).
How should I check CD209 by IF alongside the IHC result?
Use IF as a separate validation experiment and multiplex CD209 with a marker for the expected dendritic or monocyte-derived cell population (UniProt Q9NNX6 tissue specificity; HPA tissue IHC). Choose a fluorophore in a channel with low measured tissue autofluorescence, and inspect single-stain controls for bleed-through before interpreting overlap (standard IF practice). If the antibody recognises the extracellular region at residues 59–404, test staining without permeabilisation; if it recognises the cytoplasmic tail at 1–37, include an appropriately permeabilised condition (UniProt Q9NNX6 topology; standard IF practice). Keep fixation conditions documented because no directly relevant IF fixation result is supplied here (supplied evidence). Compare membrane signal and cell identity with chromogenic sections without treating the IF result as an IHC protocol (UniProt Q9NNX6 subcellular; standard IHC/IF practice).
What should I check when CD209 chromogenic staining looks diffuse?
Inspect a no-primary control to distinguish antibody-dependent signal from chromogen deposition or endogenous enzyme activity (standard chromogenic IHC practice). Apply a peroxidase block when using horseradish peroxidase detection, and assess nonspecific binding with an appropriate protein block (standard chromogenic IHC practice). Keep development time consistent across sections, because prolonged DAB development can make weak background appear cellular (standard chromogenic IHC practice). Compare questionable staining with expected selective monocyte-derived cell staining and with adipocytes, where CD209 protein was not detected in the cited tissue profile (HPA tissue IHC). Diffuse staining across unrelated compartments should prompt review of retrieval, blocking and detection controls before changing the biological interpretation (UniProt Q9NNX6 subcellular; standard IHC practice).
How should I quantify CD209 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical area before scoring, since CD209 staining is selective among monocyte-derived cells in the tissue profile (HPA tissue IHC; standard IHC practice). Report the percentage of positive eligible cells and an intensity score, or calculate an H-score from the percentage in each intensity category (standard IHC practice). For spatial comparisons, count positive cells per mm² of viable analysed tissue and state the area excluded for folds or necrosis (standard IHC practice). Normalise cell-based scores to the eligible cell population, and area-based scores to viable tissue area, rather than comparing raw positive counts between sections (standard IHC practice). Apply one positivity threshold and one chromogen development rule throughout the batch (standard IHC practice).
How do I distinguish true CD209 positivity from staining artefact?
A credible result places CD209 signal in a plausible cell population with membrane-associated staining, supported by appropriate controls (UniProt Q9NNX6 tissue specificity and subcellular; standard IHC practice). The tissue profile reports medium staining in lung macrophages and high staining in placental cytotrophoblasts, so interpret each compartment against its own morphology (HPA tissue IHC). Treat broad nuclear staining, strong section-edge signal and staining confined to necrotic material as potential artefacts requiring review (UniProt Q9NNX6 subcellular; standard IHC practice). A no-primary control helps identify endogenous enzyme or detection-system signal, particularly when DAB appears outside intact cells (standard chromogenic IHC practice). Recheck questionable fields on adjacent sections and keep unexpected cell staining separate from confirmed positives in the score (standard IHC practice).
Boster reagents
Best CD209 / CD209 antigen IHC Antibodies
The catalog antibody M01025-1 has real IHC data from human lymph node tissue (image caption); no IF image is supplied (catalog payload).
Real IHC data
Anti-DC-SIGN CD209 Monoclonal Antibody [5D7]
Cat # M01025-1
M01025-1 is listed for human IHC-P (catalog: applications and reactivity). Its IHC image caption shows human lymph node tissue stained at 5 μg/mL (image caption).
Which to pick: Choose M01025-1 for human tissue IHC: it is a mouse monoclonal, clone 5D7, listed for IHC-P, with a human lymph node IHC image (catalog: host, clone and applications; image caption). Its caption does not report the fixative (image caption). Neither M01025-1 nor A01025-2 lists IF/ICC or reactivity beyond human, so the payload supports neither an IF/ICC pick nor a cross-species pick (catalog: applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NNX6 (CD209_HUMAN, CD209 antigen).
- Human Protein Atlas. CD209 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CD209 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. CD209 antibody validation summary (2 antibodies).
- Emergence of dendritic cells in the myocardium after acute myocardial infarction - implications for inflammatory myocardial damage. International journal of biomedical science : IJBS 2010 — PMC3614732.
- Identification of Putative Biomarkers for the Early Stage of Porcine Spermatogonial Stem Cells Using Next-Generation Sequencing. PloS one 2016 — PMC4723225.
- Presence of increased inflammatory infiltrates accompanied by activated dendritic cells in the left atrium in rheumatic heart disease. PloS one 2018 — PMC6159861.
- Brain Immune Cell Composition in Multiple Sclerosis and Progressive Multifocal Leukoencephalopathy After Natalizumab: An Immunohistochemical Cohort Study. Neurology(R) neuroimmunology & neuroinflammation 2026 — PMC13262666.
- PubMed PMID:1518869 — UniProt-cited evidence.
- PubMed PMID:10975799 — UniProt-cited evidence.
- PubMed PMID:11257134 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CD209 (CD209 antigen). Expected localisation Cell membrane expected; tissue compartment unverified (UniProt; HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M01025-1 at 5 - 10 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Placenta (Cytotrophoblasts); 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 M01025-1); 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: Mouse monoclonal (clone 5D7) anti-CD209, 5 - 10 μg/mL (datasheet M01025-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Placental cytotrophoblast staining complicates DC attribution (HPA tissue IHC); Isoform / epitope: 12 isoforms; check cytoplasmic vs extracellular epitope (UniProt); Regulation: Cell-type-dependent expression (HPA tissue IHC). Evidence: UniProt Q9NNX6, HPA tissue IHC · Supported, HPA ICC-IF · Membrane, Datasheet M01025-1, PMC3614732, PMC6159861. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CD209/immunohistochemistry-ihc-guide.