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CLEC4C / C-type lectin domain family 4 member C · IHC design guide
Design Immunohistochemistry for CLEC4C
Plan chromogenic IHC on paraffin sections with the catalog antibody at 1:50 (datasheet: IHC 1:50). Interpret the observed cytoplasmic staining, most abundant in immune cells (HPA tissue IHC), alongside CLEC4C’s membrane location (UniProt) and the reported low agreement between staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CLEC4C 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 in tissue (HPA tissue IHC); cell membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining in most tissues, strongest in immune cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 | Low agreement between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Downregulated after LPS-induced maturation (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope location requires checking (UniProt) |
Section 1
Recommended CLEC4C IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with the published CLEC4C staining protocol (PMC9045650: methods).
| Sample | Paraffin-embedded human liver-cancer tissue; fixative not specified (datasheet A07313) |
| Fixation | Image fixative and duration unreported (datasheet A07313); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-CLEC4C, 1:50 (datasheet A07313) |
| 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 | CLEC4C-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, most abundant in immune cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); consider high-pH retrieval (PMC9045650: methods).
Section 2
What Is the Expected CLEC4C Staining Pattern?
CLEC4C is a cell membrane receptor with an extracellular domain spanning residues 45–213 (UniProt Q8WTT0 topology). Plasmacytoid dendritic cells are the main biological expectation (UniProt Q8WTT0 tissue specificity). HPA reports predominantly cytoplasmic staining across tissues, most abundant in immune cells; its IHC rating is Approved, with low consistency between staining and RNA expression (HPA tissue IHC).
| Discrete staining in immune cells, with membrane accentuation. | This fits CLEC4C's cell surface location and plasmacytoid dendritic cell expression (UniProt Q8WTT0). Morphology alone cannot establish plasmacytoid dendritic cell identity (general IHC practice); HPA also reports abundant immune cell staining (HPA tissue IHC). |
| Predominantly cytoplasmic staining, including glandular cells. | This resembles HPA's reported tissue pattern, including High staining in several glandular compartments (HPA tissue IHC). It conflicts with a purely surface pattern expected from UniProt topology; interpret it cautiously because HPA reports low staining–RNA consistency (HPA tissue IHC; UniProt Q8WTT0 topology). |
| Dominant nuclear staining or another compartment inconsistent with membrane or reported cytoplasmic staining. | Treat this as a possible artefact: UniProt places CLEC4C at the cell membrane, while HPA reports cytoplasmic tissue staining (UniProt Q8WTT0 subcellular location; HPA tissue IHC). Check controls and staining distribution before assigning target localisation (general IHC practice). |
| Broad staining in cells outside the expected immune population. | Do not automatically call it cross-reactivity: HPA reports High glandular staining in colon, duodenum and gallbladder (HPA tissue IHC). Compare tissue controls and cell morphology; cross-reactivity or endogenous detection activity remains possible if the pattern is inconsistent or persists in a no-primary control (general IHC practice). |
| No signal in a tissue compartment reported as strongly stained. | First check that the expected cells are present: HPA reports High staining in lung macrophages and lymph node non-germinal center cells (HPA tissue IHC). A blank result may reflect a failed staining run or antibody performance; HPA's Approved rating does not guarantee staining in every preparation (HPA tissue IHC; general IHC practice). |
💡Expected CLEC4C appearanceA credible positive is discrete immune cell staining, preferably with membrane accentuation consistent with CLEC4C topology, while diffuse slide-wide color or dominant nuclear staining warrants investigation; HPA also reports cytoplasmic immune and glandular staining, so compartment alone is not definitive (UniProt Q8WTT0; HPA tissue IHC; general IHC practice).
| Epitope position | CLEC4C has a cytoplasmic region at residues 1–21, a transmembrane segment at 22–44 and an extracellular region at 45–213 (UniProt Q8WTT0 topology). The supplied record does not locate the IHC antibody's epitope, so it cannot predict its section staining pattern. |
| Glycosylation and variants | UniProt lists glycosylation sites at 110, 137 and 164 and 2 isoforms (UniProt Q8WTT0). These facts do not establish whether this antibody distinguishes isoforms or whether glycosylation changes IHC staining; do not use them to explain a weak result without antibody-specific evidence. |
| Cell population and maturation | UniProt reports expression in plasmacytoid dendritic cells and immature monocyte-derived dendritic cells, with down-regulation after LPS-driven maturation but not TNF treatment (UniProt Q8WTT0 tissue specificity). Cell composition can therefore affect the biological expectation; the supplied HPA IHC data do not identify every stained immune cell subtype (HPA tissue IHC). |
| Tissue IHC evidence | HPA rates the listed antibody Approved and describes low consistency between antibody staining and RNA expression; it does not list an IHC Enhanced rating (HPA tissue IHC; HPA antibody validation). Use its High and Low calls as observed staining levels, not proof of cell-specific expression or validated negative controls. |
| Antigen retrieval | Retrieval is a general paraffin IHC variable to optimize with appropriate controls (general IHC practice). No supplied source reports a CLEC4C-specific retrieval condition or fixation sensitivity; neither UniProt topology nor HPA staining levels justify predicting a target-specific retrieval effect. |
| IF/ICC Q&A | Q: Should IF/ICC show membrane signal? A: Membrane localisation is the biological expectation (UniProt Q8WTT0; HPA subcellular summary), but HPA supplies no ICC-IF images or main-location assignment for CLEC4C (HPA subcellular). Interpret any IF pattern on its own controls; no IF protocol follows from these records. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in lung macrophages or lymph node non-germinal center cells. | The expected cells may be absent from the section, or the IHC run may have failed (HPA tissue IHC; general IHC practice). | Confirm the cell compartment on the counterstained section and inspect a known-performing run control; then review retrieval, primary application and detection steps as general IHC checks (general IHC practice). |
| Diffuse color across tissue and blank areas. | Background from detection reagents, inadequate blocking or washing is possible (general IHC practice). | Compare a no-primary control and reagent-only background, then adjust blocking or washes according to the detection system (general IHC practice); diffuse color is not a documented CLEC4C localisation (UniProt Q8WTT0; HPA tissue IHC). |
| Strong staining of unexpected cell types. | It may reflect HPA's broad cytoplasmic pattern, cross-reactivity or endogenous detection activity; HPA reports low staining–RNA consistency (HPA tissue IHC; general IHC practice). | Check whether the cell type appears in HPA's tissue observations, compare tissue controls and examine a no-primary control before assigning CLEC4C expression (HPA tissue IHC; general IHC practice). |
| Nuclear staining dominates. | This differs from UniProt's membrane location and HPA's reported cytoplasmic IHC profile (UniProt Q8WTT0; HPA tissue IHC). | Treat nuclear signal as unconfirmed; inspect controls and review counterstain and detection background before scoring positive cells (general IHC practice). |
| Only weak staining in oral mucosa or splenic red pulp. | HPA lists oral squamous epithelial cells and splenic red pulp cells as Low, rather than negative (HPA tissue IHC). | Avoid using either compartment as an absolute negative control; compare with an HPA High compartment and score the observed cell type separately (HPA tissue IHC; general IHC practice). |
| Predominantly cytoplasmic signal despite an expected surface receptor. | HPA explicitly describes cytoplasmic tissue staining, while UniProt places CLEC4C at the membrane (HPA tissue IHC; UniProt Q8WTT0). | Record the compartment and cell type separately, compare controls and report the source disagreement; do not infer a CLEC4C-specific fixation or retrieval mechanism from these records. |
Experiment evidence
IHC & IF images showing CLEC4C staining
A07313 · IHCImmunohistochemistry validation of CLEC4C using Anti-CD303 CLEC4C Antibody (A07313). Immunohistochemical analysis of paraffin-embedded human
A07313 · IHCImmunohistochemistry validation of CLEC4C using Anti-CD303 CLEC4C Antibody (A07313). Immunohistochemical analysis of paraffin-embedded human
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198178-CLEC4C/tissue/cerebral+cortex#imid_7227776
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198178-CLEC4C/tissue/colon#imid_7227710
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198178-CLEC4C/tissue/liver#imid_7227695
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198178-CLEC4C/tissue/kidney#imid_7227696
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CLEC4C IHC & IF
🧪Run colon first and assess its glandular cells for staining (HPA: High in colon glandular cells). HPA lists no negative tissue and detects CLEC4C in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; non-glandular cells on the colon slide can be checked for background but are not established negative cells (HPA: no negative rows; 45 tissues detected).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: CLEC4C is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CLEC4C; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a secondary-only slide with no primary antibody, an isotype control matched to the primary antibody’s host species and antibody type, and, if available, a knockout specimen or immunizing-peptide competition control (standard IHC practice). For colon sections, quench endogenous peroxidase and inspect glandular cells for residual chromogenic background (HPA: High in colon glandular cells; standard chromogenic IHC practice).
⚠️Feasibility: No supplied target or application evidence reports a CLEC4C-specific fixation window or antigen-retrieval dependence; establish retrieval conditions empirically (supplied evidence: no fixation-window or retrieval data). The selected A07313 caption describes paraffin-embedded human liver cancer, but its fixative is unreported (A07313 caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier; in colon, assess peroxidase background when interpreting the expected cell-membrane localization (HPA: High in colon glandular cells; UniProt Q8WTT0: cell membrane).
HPA tissue IHC evidence for CLEC4C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CLEC4C is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CLEC4C IHC Tips
These questions focus on CLEC4C staining in paraffin sections, with one entry on IF multiplexing (selected IHC caption; UniProt Q8WTT0).
What retrieval should I start with for CLEC4C in paraffin sections?
For CLEC4C paraffin IHC, start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Allow sections to cool in retrieval buffer, then compare staining with a section processed without retrieval to assess whether heating improves the signal (standard IHC practice). If staining is weak, test a higher-pH retrieval buffer on adjacent sections while holding detection and exposure to chromogen constant (standard IHC practice). Score cell-associated staining and tissue damage together; the selected antibody has a paraffin-section image, but its caption gives no retrieval conditions (A07313 caption).
How should I handle fixation when CLEC4C staining is weak?
The selected antibody was shown on a paraffin-embedded human liver cancer section, but the caption does not state its fixative or fixation duration (A07313 caption). CLEC4C-specific sensitivity to fixation is therefore unknown; avoid attributing weak staining to a particular fixation effect without a controlled comparison (A07313 caption). For new specimens, record fixative, fixation duration and processing schedule, and compare adjacent sections under the same retrieval and detection conditions (standard IHC practice). Include a documented positive tissue section in each run, then assess whether signal loss tracks processing history or affects both test and control sections (standard IHC practice).
Should CLEC4C stain the membrane or cytoplasm in tissue IHC?
CLEC4C is a cell-membrane protein with a cytoplasmic segment at residues 1–21, a transmembrane segment at 22–44 and an extracellular region at 45–213 (UniProt Q8WTT0 topology). Examine the cell perimeter at high power and document whether any cytoplasmic signal accompanies it (UniProt Q8WTT0 topology; standard IHC practice). HPA reports predominantly cytoplasmic tissue staining, while its tissue assessment notes low consistency between staining and RNA expression (HPA tissue IHC). Treat diffuse cytoplasmic staining as requiring corroboration with cell identity and controls before assigning it to CLEC4C (HPA tissue IHC; standard IHC practice).
How do isoforms and epitope position affect my CLEC4C IHC readout?
UniProt lists 2 CLEC4C isoforms, but the supplied antibody caption does not identify its epitope or establish which isoforms it detects (UniProt Q8WTT0 isoforms; A07313 caption). The recorded topology places residues 1–21 inside the cell and 45–213 outside it (UniProt Q8WTT0 topology). The extracellular lectin domain spans 90–207 and has glycosylation sites at 110, 137 and 164; these features do not locate this antibody’s epitope (UniProt Q8WTT0). If staining changes with retrieval, compare conditions on adjacent sections and avoid assigning isoform specificity or glycan sensitivity without direct epitope evidence (standard IHC practice; A07313 caption).
How can I check a CLEC4C IHC finding with multiplex IF?
For an independent check, pair CLEC4C with a validated plasmacytoid dendritic-cell marker and score overlap in individual cells (UniProt Q8WTT0 tissue specificity; standard IF practice). Choose a far-red channel for a weak CLEC4C signal when tissue autofluorescence is strongest at shorter wavelengths, and inspect single-stain controls for bleed-through (standard IF practice). An extracellular epitope may be accessible without permeabilisation, whereas an epitope in residues 1–21 requires access to the cytoplasmic side; this antibody’s epitope is unspecified (UniProt Q8WTT0 topology; A07313 caption). Optimise permeabilisation against the actual epitope and specimen, then compare the IF cell pattern with the chromogenic IHC result (standard IF practice).
What should I check when CLEC4C DAB staining looks widespread?
First compare no-primary and isotype-appropriate control sections with the test section to identify staining from detection reagents or nonspecific binding (standard IHC practice). Apply a peroxidase block before DAB detection and inspect whether pigment, blood-rich areas or tissue edges retain color in controls (standard chromogenic IHC practice). HPA describes cytoplasmic staining across many tissues but also reports low consistency between antibody staining and RNA expression, so widespread color alone is insufficient evidence (HPA tissue IHC). Check whether the signal follows identifiable cells and the expected membrane distribution, then adjust blocking or antibody concentration only against a stable positive control (UniProt Q8WTT0 subcellular location; standard IHC practice).
How should I quantify CLEC4C-positive cells across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis compartment and tissue region before counting, and exclude folds, necrosis and areas with failed counterstaining (standard IHC practice). For cell prevalence, report CLEC4C-positive cells as a percentage of evaluable nucleated cells; for spatial abundance, report positive-cell density per mm² of viable tissue (standard IHC practice). If staining intensity is reproducible, calculate an H-score from the percentages at prespecified intensity grades and retain the underlying cell counts (standard IHC practice). Normalise comparisons to the same cell population or viable area, and report membranous and cytoplasmic scores separately because the recorded protein location and HPA tissue pattern differ (UniProt Q8WTT0 subcellular location; HPA tissue IHC).
When is a CLEC4C-positive IHC result convincing?
A convincing result is cell-associated, reproducible across comparable sections and consistent with CLEC4C’s recorded cell-membrane location (UniProt Q8WTT0 subcellular location; standard IHC practice). Consider the cell identity: UniProt reports expression in plasmacytoid dendritic cells, while HPA lists several strongly stained nonimmune cell types and flags low staining-to-RNA consistency (UniProt Q8WTT0 tissue specificity; HPA tissue IHC). Recheck staining confined to cut edges, necrotic regions or endogenous enzyme-rich areas against matched controls before calling it positive (standard chromogenic IHC practice). When compartment or cell type conflicts with expectations, seek corroboration with an independent marker or assay and report the observed pattern without assigning unverified specificity (standard IHC practice; UniProt Q8WTT0).
Boster reagents
Best CLEC4C / C-type lectin domain family 4 member C IHC Antibodies
A07313 is a human-reactive anti-CLEC4C antibody with an IHC image from a paraffin-embedded human liver cancer sample (catalog: reactivity; A07313 IHC caption). IF is listed, but no IF image is supplied (catalog: applications; image alts).
Real IHC data
Anti-CD303 CLEC4C Antibody
Cat # A07313
A07313 will render with an IHC figure from paraffin-embedded human liver cancer (A07313 IHC caption). Its listed applications include IHC and IF, with human reactivity; no IF figure is supplied (catalog: applications, reactivity, image alts).
Which to pick: Choose A07313 for human tissue IHC: its own image caption documents staining of a paraffin-embedded human liver cancer sample, but does not report the fixative (A07313 IHC caption). For IF/ICC, A07313 lists IF and human reactivity, although the payload provides no IF image or ICC validation; its host is rabbit and clonality is unspecified (catalog: applications, reactivity, host, clone, image alts). No cross-species option is documented because A07313 lists human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8WTT0 (CLC4C_HUMAN, C-type lectin domain family 4 member C).
- Human Protein Atlas. CLEC4C tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CLEC4C subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. CLEC4C antibody validation summary (1 antibodies).
- Stress-driven reprogramming of plasmacytoid dendritic cells in intrahepatic cholangiocarcinoma defines a reversible targetable immunosuppressive state. World journal of surgical oncology 2026 — PMC13589389.
- Human macrophages and monocyte-derived dendritic cells stimulate the proliferation of endothelial cells through midkine production. PloS one 2022 — PMC9045650.
- PubMed PMID:11536172 — UniProt-cited evidence.
- PubMed PMID:11748283 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CLEC4C (C-type lectin domain family 4 member C). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); cell membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07313 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver-cancer tissue; fixative not specified (datasheet A07313); 2. Fixation: Image fixative and duration unreported (datasheet A07313); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CLEC4C, 1:50 (datasheet A07313); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Low agreement between staining and RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope location requires checking (UniProt); Regulation: Downregulated after LPS-induced maturation (UniProt). Evidence: UniProt Q8WTT0, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A07313, PMC9045650. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CLEC4C/immunohistochemistry-ihc-guide.