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CD83 / CD83 antigen · IHC design guide
Design Immunohistochemistry for CD83
Plan CD83 IHC around cytoplasmic staining in subsets of immune cells across several tissues (HPA tissue IHC). Use consistent fixation and interpret staining with HPA’s reported off-target binding in mind (standard IHC practice; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CD83 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 | Cytoplasm in immune-cell subsets (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in subsets of immune cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+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 | Presumed off-target staining may complicate interpretation (HPA tissue IHC) | |
| Regulation | Expressed by activated immune cells (UniProt) | |
| Isoform / epitope | No isoforms reported; extracellular vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended CD83 IHC & IF Protocols
The catalog antibody protocol is paired with 3 published CD83 IHC protocols from paraffin sections (PMC6281009; PMC11554423; PMC4433839).
| Sample | Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A01777) |
| Fixation | Image fixative and duration unreported (datasheet A01777); 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-CD83, 1:50-1:200 (datasheet A01777) |
| 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 | CD83-positive staining in purkinje cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a subsets of immune cells in several tissues, including lymphoid organs. Additional expression in brain. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); CD83 is a membrane protein (UniProt Q01151).
Section 2
What Is the Expected CD83 Staining Pattern?
CD83 is a transmembrane protein with an extracellular region at residues 20–144 and a cytoplasmic region at 167–205 (UniProt Q01151 topology). In paraffin tissue sections, expect staining in subsets of immune cells, including cells in lymphoid organs (HPA tissue IHC: Enhanced reliability; medium staining consistency with RNA). UniProt also reports expression in activated lymphocytes, Langerhans cells and activated dendritic cells (UniProt Q01151 tissue specificity).
| Discrete staining in immune-cell subsets, with membrane and/or cytoplasmic signal. | This fits the HPA tissue IHC profile, which describes cytoplasmic staining in subsets of immune cells, and the membrane location reported by UniProt (HPA tissue IHC; UniProt Q01151 subcellular location). Score the stained cell population and compartment together; the HPA profile does not imply that every immune cell must stain. |
| Strong staining in cerebellar Purkinje cells. | This can be an expected tissue-specific result: HPA reports high cytoplasmic/membrane staining in Purkinje cells (HPA tissue IHC: cerebellum). It should not automatically be classified as an off-target result because CD83 is also associated with immune cells (HPA tissue IHC; UniProt Q01151 tissue specificity). |
| Predominantly nuclear staining in a tissue section, without the expected cell pattern. | Treat this as unconfirmed and investigate staining artefact. HPA ICC-IF lists nucleoplasm as an uncertain location, while plasma membrane is approved; its tissue IHC summary describes cytoplasmic immune-cell staining (HPA subcellular; HPA tissue IHC). An ICC-IF nuclear observation alone does not validate nuclear IHC staining. |
| Widespread staining of bronchial respiratory epithelium or another unexpected cell population. | Check for cross-reactivity or endogenous detection activity before assigning CD83 expression (general IHC practice). HPA reports bronchial respiratory epithelial cells as not detected and notes presumed off-target binding in its tissue IHC assessment (HPA tissue IHC: bronchus; HPA reliability description). |
| Diffuse staining across the section, or no signal in a known-positive area. | Diffuse background prevents a confident cell-level call; absent signal requires a control and workflow check (general IHC practice). For comparison, HPA reports medium staining in tonsillar germinal-center cells and lung macrophages (HPA tissue IHC: tonsil, lung). A negative run cannot overturn those observations without a working positive control. |
💡Expected CD83 appearanceCall an IHC result positive when staining is confined to an appropriate cell population and shows a discernible membrane and/or cytoplasmic pattern: often medium in reported immune-cell sites, or high in cerebellar Purkinje cells (HPA tissue IHC; UniProt Q01151 topology); widespread signal in cells reported as not detected is suspect (HPA tissue IHC: bronchus).
| Cell type and tissue context | HPA reports medium staining in appendix and tonsil germinal-center cells, bone-marrow hematopoietic cells, lung macrophages, lymph-node non-germinal-center cells and splenic red-pulp cells (HPA tissue IHC). Interpret a positive result by identifying the stained cells, since signal intensity alone does not establish specificity (general IHC practice). |
| Topology and unknown antibody epitope | CD83 has an extracellular region at 20–144, a transmembrane segment at 145–166 and a cytoplasmic region at 167–205 (UniProt Q01151 topology). The supplied record does not locate the antibody epitope; select any permeabilisation decision using the antibody's stated epitope and assay instructions (general IHC practice). |
| Antigen retrieval | Retrieval conditions can affect immunostaining in paraffin sections (general IHC practice). No CD83-specific retrieval condition or fixation sensitivity is established by the supplied UniProt and HPA records. Use the IHC-validated antibody's documented conditions as a starting point and judge changes against positive and negative controls (general IHC practice). |
| Can ICC-IF localisation guide this IHC call? | Use it as context, not a paraffin-section protocol: HPA ICC-IF approves plasma membrane and Golgi localisation, while nucleoplasm and nucleoli are uncertain (HPA subcellular). Compare those findings with the cytoplasmic immune-cell tissue IHC profile when interpreting a section (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in an expected positive area. | The run may have failed, or the sampled section may lack the relevant cells (general IHC practice). | Confirm that the expected population is present; use a positive tissue control with a reported HPA pattern, such as tonsillar germinal-center cells, and check the antibody's IHC conditions and detection steps (HPA tissue IHC: tonsil; general IHC practice). |
| Weak or patchy staining in lymphoid tissue. | CD83 staining can be limited to a subset of cells; HPA rates tonsillar germinal-center cells medium, not uniformly high (HPA tissue IHC: tonsil; HPA tissue profile). | Inspect the relevant cell compartment and compare it with a concurrent positive control before calling the section negative (general IHC practice). Record cell distribution separately from stain intensity so a small positive subset remains visible in the interpretation. |
| Broad epithelial staining, including bronchial respiratory epithelium. | Cross-reactivity or endogenous detection activity may mimic a positive stain (general IHC practice). Bronchial respiratory epithelial cells are reported as not detected (HPA tissue IHC: bronchus). | Review a no-primary control for detection background and compare staining with an independently validated IHC antibody if available (general IHC practice). HPA lists two IHC antibodies with Enhanced validation, HPA041454 and CAB002509 (HPA antibodies). |
| Section-wide haze obscures individual cells. | Excess nonspecific antibody or detection background can obscure cell boundaries (general IHC practice). | Check the no-primary control, blocking and washes; adjust antibody concentration only within the chosen antibody's documented IHC conditions (general IHC practice). Reassess whether distinct cells and compartments emerge before scoring CD83. |
| Nuclear-only staining is being scored as CD83 positive. | Nucleoplasm is an uncertain ICC-IF location, and nuclear-only staining does not match the HPA tissue IHC summary of cytoplasmic immune-cell subsets (HPA subcellular; HPA tissue IHC). | Recheck the stained cell type, counterstain and controls, then compare with a section showing the reported tissue pattern (general IHC practice; HPA tissue IHC). Keep the nuclear result unconfirmed unless independent evidence supports it. |
| The proposed negative control tissue contains some stained cells. | A tissue-level label can hide cell-specific expression; HPA reports both positive and not-detected cell populations by tissue (HPA tissue IHC). | Score the specified negative population rather than treating every cell in that tissue as negative. For example, adipocytes in adipose tissue are reported not detected (HPA tissue IHC: adipose tissue); assess unexpected staining against that cell-level reference. |
Experiment evidence
IHC & IF images showing CD83 staining
A01777 · IHCImmunohistochemistry (IHC) analyzes of CD83 (P130) pAb in paraffin-embedded human lung carcinoma tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000112149-CD83/tissue/cerebral+cortex#imid_815024
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000112149-CD83/tissue/cerebellum#imid_815019
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000112149-CD83/tissue/colon#imid_814942
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000112149-CD83/tissue/liver#imid_814943
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA064421; green = CD83, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA064421; green = CD83, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CD83 IHC & IF
🧪Run cerebellum first: Purkinje cells should show cytoplasmic or membrane staining (HPA: High in Purkinje cells). Use adipose tissue as the negative tissue because adipocytes are not detected (HPA: Not detected in adipocytes); on the cerebellar slide, compare other cells with Purkinje cells for background, without assuming every other cell is CD83-negative (HPA: High in Purkinje cells).
Positive control tissue: Cerebellum (Purkinje cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CD83 in HeLa, Rh30, U2OS, with annotated localisation: Nucleoplasm (uncertain), Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and, where available, a CD83 knockout or peptide-block control (standard IHC practice). Quench endogenous peroxidase for chromogenic IHC, and assess neuronal autofluorescence if using IF on cerebellum (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A01777 paraffin-section caption also leaves the fixative unreported (selected IHC caption: paraffin-embedded lung carcinoma; fixative not stated). Optimize antigen retrieval empirically for paraffin IHC; the supplied evidence does not establish that frozen sections or IF are easier for CD83 (standard IHC practice). In cerebellar IF, neuronal autofluorescence can complicate interpretation, so check an unstained section (standard IF practice).
HPA tissue IHC evidence for CD83
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded. 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Appendix | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Lymph node | Non-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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CD83 IHC Tips
Troubleshoot CD83 staining in paraffin sections by checking retrieval, cellular context, and compartment before interpreting chromogenic signal.
How should I retrieve CD83 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval specification). Allow sections to cool in retrieval buffer, then compare staining with an otherwise identical section processed without retrieval; this isolates the effect of heating (standard IHC practice). If staining remains weak, titrate retrieval duration on adjacent sections before testing another buffer or pH, while watching for tissue damage and increased background (standard IHC practice). Assess signal in immune cells alongside morphology, since CD83 is reported in activated dendritic cells and lymphocytes (UniProt Q01151 tissue specificity).
Could fixation explain weak or uneven CD83 staining?
Target-specific fixation sensitivity for CD83 is unknown from the supplied evidence, so record the actual fixative, fixation duration, and processing history before changing the staining workflow (supplied evidence; standard IHC practice). The catalog image shows paraffin-embedded human lung carcinoma stained with antibody A01777, but its caption does not state a fixative (catalog caption A01777). Compare sections with similar processing histories and keep retrieval at citrate pH 6.0, 95–98 °C for 20 min while testing other variables (page retrieval specification; standard IHC practice). If signal varies across a section, inspect preservation and staining controls before attributing the pattern to CD83 biology (standard IHC practice).
Should CD83 staining appear at the membrane or in the cytoplasm?
Prioritize a convincing cell-associated membrane pattern: CD83 has an extracellular region at residues 20–144, one transmembrane segment at 145–166, and a cytoplasmic tail at 167–205 (UniProt Q01151 topology). Cytoplasmic staining can occur in tissue IHC, and the tissue atlas reports it in subsets of immune cells across several tissues (HPA tissue IHC profile). For interpretation, compare the chromogenic deposit with cell borders, cytoplasm, and a hematoxylin counterstain rather than assigning faint diffuse color to a compartment (standard IHC practice). The subcellular atlas approves plasma membrane and Golgi localisation, while its nucleoplasmic and nucleolar assignments are uncertain (HPA subcellular).
How does epitope location affect CD83 staining in paraffin sections?
Map the antibody's stated immunogen or epitope before interpreting a negative section; the supplied record lists 0 isoforms and does not specify this antibody's epitope (UniProt Q01151 isoforms; supplied antibody evidence). An extracellular epitope would lie within residues 20–144, whereas a tail epitope would lie within 167–205 (UniProt Q01151 topology). The extracellular region includes an Ig-like domain at 20–114 and glycosylation sites at 79, 96, and 117, so epitope accessibility should be assessed experimentally rather than assumed (UniProt Q01151 domain and glycosylation; standard IHC practice). Compare retrieval conditions on matched sections and judge any gain against morphology and background (standard IHC practice).
How can IF help verify an ambiguous CD83 IHC pattern?
Use IF as a separate cross-check, multiplexing CD83 with a validated marker of the immune cell population being evaluated; CD83 is reported in activated dendritic cells and lymphocytes (UniProt Q01151 tissue specificity; standard IF practice). Choose fluorophores after examining unstained tissue in each channel, favoring a detection channel with low tissue autofluorescence and using single-stain controls to check spectral spillover (standard IF practice). For an antibody against the extracellular region at residues 20–144, compare staining without permeabilisation first; a cytoplasmic-tail epitope at 167–205 requires access across the membrane (UniProt Q01151 topology; standard IF practice). Interpret IF membrane signal alongside IHC morphology, since the subcellular atlas approves plasma membrane localisation (HPA subcellular).
What should I check when CD83 chromogenic staining is widespread?
Start with a no-primary control and inspect whether color persists in the same structures; persistent deposit points to the detection workflow or tissue background (standard IHC practice). For peroxidase-based detection, check the peroxidase block and compare sections before and after DAB development, keeping development time constant across the run (standard IHC practice). Titrate the primary antibody and strengthen appropriate blocking or washes if diffuse staining obscures cell boundaries (standard IHC practice). Be cautious with widespread signal because the tissue atlas reports presumed off-target binding, and its CD83 staining has medium consistency with RNA expression (HPA tissue IHC reliability).
How should I quantify CD83-positive cells in tissue IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the sampled compartment and cell population before scoring, using morphology and a consistent hematoxylin counterstain to distinguish cells from extracellular deposit (standard IHC practice). Report CD83-positive cells as a percentage of eligible cells or a density per mm²; if intensity matters, add an H-score from intensity categories 0–3 multiplied by their percentages (standard IHC practice). Normalize counts to the same tissue area or eligible-cell denominator across sections, and exclude necrosis, folds, and edges by a predefined rule (standard IHC practice). Keep membrane and cytoplasmic scores separate because CD83 is a membrane protein but tissue IHC also reports cytoplasmic immune-cell expression (UniProt Q01151 subcellular; HPA tissue IHC profile).
How can I distinguish genuine CD83 staining from artefact?
A credible result should fit identifiable cells and a plausible compartment: CD83 is a membrane protein expressed by activated dendritic cells and lymphocytes, while tissue IHC also reports cytoplasmic staining in immune-cell subsets (UniProt Q01151 subcellular and tissue specificity; HPA tissue IHC profile). Compare staining with morphology and a no-primary control, and scrutinize section edges, necrotic areas, and diffuse extracellular color before scoring (standard IHC practice). Color that remains without primary antibody warrants checking endogenous peroxidase blocking and detection reagents (standard IHC practice). Interpret unexpected cell types cautiously because the tissue atlas notes presumed off-target binding and only medium consistency between staining and RNA expression (HPA tissue IHC reliability).
Boster reagents
Best CD83 / CD83 antigen IHC Antibodies
The catalog has one anti-CD83 antibody with a real IHC image from paraffin-embedded human lung carcinoma tissue (image caption); Human, Mouse and Rat reactivity is listed (catalog). No IF data is shown (catalog).
Real IHC data
Anti-CD83 (P130) Antibody
Cat # A01777
A01777 is listed for IHC and WB, with Human, Mouse and Rat reactivity (catalog). Its IHC image shows paraffin-embedded human lung carcinoma tissue (image caption); no IF image is provided (catalog).
Which to pick: For paraffin-section IHC, choose A01777: its own image caption documents staining in paraffin-embedded human lung carcinoma tissue, and the listed IHC dilution is 1:50–1:200 (image caption; datasheet). The caption does not report the fixative (image caption). For Mouse or Rat tissue, A01777 lists reactivity, but its IHC image documents only human tissue (catalog; image caption); no IF/ICC application or IF dilution is listed, so there is no IF/ICC pick here (catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q01151 (CD83_HUMAN, CD83 antigen).
- Human Protein Atlas. CD83 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CD83 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and plasma membrane. In addition localized to the nucleoli and the Golgi apparatus..
- Human Protein Atlas. CD83 antibody validation summary (3 antibodies).
- Inflammatory cell expression of Toll-like receptor-2 (TLR2) within refractory periapical granuloma. F1000Research 2018 — PMC6281009.
- Analysis of CD83 antigen expression in human breast fibroadenoma and adjacent tissue. Sao Paulo medical journal = Revista paulista de medicina 2011 — PMC10868929.
- An Immunohistochemical Study on the Role of CD83+ Dendritic Cells (DCs) in Malignant and Benign Lesions of the Human Cervix. Cureus 2024 — PMC11554423.
- VEGF expression, microvessel density and dendritic cell decrease in thyroid cancer. Biotechnology, biotechnological equipment 2014 — PMC4433839.
- PubMed PMID:1378080 — UniProt-cited evidence.
- PubMed PMID:8422464 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CD83 (CD83 antigen). Expected localisation Cytoplasm in immune-cell subsets (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A01777 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells - cytoplasm/membrane); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung carcinoma tissue; fixative not specified (datasheet A01777); 2. Fixation: Image fixative and duration unreported (datasheet A01777); 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-CD83, 1:50-1:200 (datasheet A01777); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining may complicate interpretation (HPA tissue IHC); Isoform / epitope: No isoforms reported; extracellular vs cytoplasmic epitope matters (UniProt); Regulation: Expressed by activated immune cells (UniProt). Evidence: UniProt Q01151, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm and plasma membrane. In addition…, Datasheet A01777, PMC6281009, PMC11554423. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CD83/immunohistochemistry-ihc-guide.