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CD83 / CD83 antigen · IHC design guide
Design Immunohistochemistry for CD83
Plan CD83 IHC on paraffin sections using tissue staining examples and controls. Expect cytoplasmic staining in subsets of immune cells (HPA tissue IHC); CD83 is a membrane protein with an extracellular domain (UniProt).
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 | Cytoplasmic staining 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining was observed and disregarded (HPA tissue IHC) | |
| Regulation | Expressed by activated lymphocytes and dendritic cells (UniProt) | |
| Isoform / epitope | No isoforms; epitope side matters: residues 20–144 vs 167–205 (UniProt) |
Section 1
Recommended CD83 IHC & IF Protocols
The catalog antibody protocol is accompanied by published CD83 IHC methods for periapical, breast, cervical and thyroid specimens (PMC6281009; PMC10868929; 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 HIER at 95–98 °C for 20 min (page retrieval); compare the published tissue methods if staining needs optimization.
Section 2
What Is the Expected CD83 Staining Pattern?
CD83 is a surface glycoprotein with an extracellular region at residues 20–144 and a transmembrane segment at 145–166 (UniProt Q01151 topology). In paraffin-section IHC, expect staining in subsets of immune cells, often cytoplasmic by HPA observation (HPA tissue IHC). Interpret positives cautiously: HPA rates the tissue profile Enhanced but reports medium agreement with RNA data and disregarded presumed off-target staining (HPA tissue IHC).
| Discrete membrane-associated or cytoplasmic staining in scattered immune cells within lymphoid tissue. | This fits CD83 surface localization and HPA's observed cytoplasmic staining in immune-cell subsets (UniProt Q01151 topology; HPA tissue IHC). HPA reports medium staining in appendix and tonsil germinal-center cells, lymph-node non-germinal-center cells, and splenic red-pulp cells (HPA tissue IHC). Score the stained cells and compartment, rather than calling the entire tissue positive. |
| Strong, widespread nuclear staining dominates a paraffin section. | Reassess specificity and detection background before accepting this as CD83 IHC: UniProt places CD83 at the membrane, while HPA's nuclear ICC-IF locations are marked uncertain (UniProt Q01151 subcellular location; HPA subcellular ICC-IF). HPA does report cytoplasmic tissue staining, so cytoplasm alone is not a reason to reject a result (HPA tissue IHC). |
| Chromogen appears in cells expected to lack CD83, such as adipocytes. | HPA did not detect staining in adipocytes in adipose tissue or breast (HPA tissue IHC). Check whether the signal follows cell boundaries and whether negative-reagent controls also stain; cross-reactivity or endogenous detection activity are possible explanations (general IHC practice). Do not treat every cell in an otherwise positive organ as a CD83-positive cell. |
| A diffuse haze covers cells, stroma, or the whole section without a discernible cellular pattern. | This cannot be scored as the discrete immune-cell staining HPA describes (HPA tissue IHC). Review blocking, antibody concentration, washing, and chromogen development using matched controls (general IHC practice). HPA notes presumed off-target binding in its tissue assessment, so diffuse staining needs independent scrutiny (HPA tissue IHC). |
| A run shows no cellular staining in an expected-positive control section. | Check the section and IHC run before interpreting study samples as CD83-negative (general IHC practice). HPA reports medium staining in tonsil germinal-center cells and bone-marrow hematopoietic cells; neither designation guarantees staining in every specimen or cell (HPA tissue IHC). Verify that the chosen control contains the relevant cell population. |
💡Expected CD83 appearanceCall a result positive when discrete membrane-associated or cytoplasmic staining marks the expected immune-cell subset, typically at HPA's reported medium level in listed lymphoid tissues; widespread nuclear or structureless staining warrants review (UniProt Q01151 topology; HPA tissue IHC; HPA subcellular ICC-IF).
| Cell population and tissue context | CD83 is expressed by activated lymphocytes, Langerhans cells, and activated dendritic cells (UniProt Q01151 tissue specificity). HPA observes immune-cell subsets across several tissues, including lymphoid organs, and also reports staining in Purkinje and Leydig cells (HPA tissue IHC). Identify the stained population before comparing sections. |
| Epitope position and mature protein | The signal peptide spans residues 1–19; the annotated mature chain spans 20–205, with extracellular residues 20–144 (UniProt Q01151 processing and topology). An antibody's epitope would determine which region it recognizes (general antibody practice). No epitope or CD83-specific retrieval condition is supplied here, so do not infer one. |
| Glycosylated extracellular region | UniProt lists glycosylation sites at residues 79, 96, and 117 within the extracellular region (UniProt Q01151 glycosylation and topology). This describes the antigen, but does not establish how fixation or retrieval affects the IHC signal for a particular antibody. |
| Strength of tissue evidence | HPA labels the tissue IHC profile Enhanced and lists two antibodies with Enhanced IHC validation, HPA041454 and CAB002509 (HPA tissue IHC; HPA antibodies). HPA also reports medium consistency with RNA data and presumed off-target staining that it disregarded; assess unexpected cells independently (HPA tissue IHC). |
| IF/ICC interpretation | For the separate IF/ICC application, HPA approves plasma-membrane and Golgi localization but marks nucleoplasm and nucleoli uncertain (HPA subcellular ICC-IF). Those cell-image observations do not establish a paraffin-section IHC pattern or an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected immune-cell staining is absent in the positive-control section. | The relevant cells may be missing from the section, or an IHC run step may have failed (general IHC practice). HPA's medium tissue level is an observation, not a guaranteed signal in every section (HPA tissue IHC). | Confirm the control contains the annotated cell population, then review reagent omission controls, antibody dilution, detection, and chromogen development according to the validated IHC workflow (general IHC practice). |
| Nuclear staining is stronger than membrane or cytoplasmic staining. | Dominant nuclear staining has weak support for tissue IHC: CD83 is membrane-associated, and HPA marks its nuclear ICC-IF locations uncertain (UniProt Q01151 subcellular location; HPA subcellular ICC-IF). | Compare cellular morphology and negative-reagent controls, and repeat with an independently IHC-validated antibody if available (general IHC practice; HPA antibodies). Keep the uncertain IF nuclear observation separate from IHC scoring. |
| Adipocytes or another HPA-negative cell population stains. | The finding conflicts with HPA's absence of adipocyte staining; cross-reactivity or endogenous detection activity may contribute (HPA tissue IHC; general IHC practice). Nearby immune cells require separate assessment. | Inspect cell identity, run appropriate negative-reagent and endogenous-activity controls, and compare staining with the expected immune-cell distribution (general IHC practice; HPA tissue IHC). |
| A broad brown haze obscures individual positive cells. | Nonspecific reagent binding, inadequate washing, or excess chromogen development can produce diffuse background (general IHC practice). HPA reports disregarded presumed off-target staining in its tissue assessment (HPA tissue IHC). | Review blocking, dilution, washes, and development against control sections; score only a resolvable cellular pattern (general IHC practice). |
| A lymphoid section appears uniformly positive. | HPA describes staining in subsets, with particular compartments reported in appendix, tonsil, lymph node, and spleen (HPA tissue IHC). Uniform color may reflect background or an overbroad cell call (general IHC practice). | Score the named cell compartment and staining location at higher magnification, then compare with negative-reagent controls (HPA tissue IHC; general IHC practice). |
| Does nucleoplasmic IF/ICC staining confirm a positive CD83 IHC section? | HPA marks nucleoplasm uncertain in ICC-IF, while plasma membrane is approved; the applications provide different observations (HPA subcellular ICC-IF). | Use the IF/ICC finding only with its stated uncertainty. Judge paraffin-section IHC against its own cellular pattern and controls (HPA tissue IHC; general IHC practice). |
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 and expect Purkinje cells to stain (HPA: High in Purkinje cells). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); neighboring cells without specific staining on the cerebellum slide should remain at background level.
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, a nonimmune isotype control matched to the catalog antibody’s host species and polyclonal format, and CD83-knockout material as a biological negative (selected A01777 caption: pAb). For chromogenic cerebellum IHC, block endogenous peroxidase and check pigment against the control slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of fixation on CD83 staining are unreported in the supplied evidence; the fixative in the selected paraffin-section caption is also unreported (selected A01777 caption: paraffin-embedded tissue; fixative not stated). Retrieval dependence and whether frozen sections or IF are easier are unreported, so optimize antigen retrieval on paraffin sections before scoring (standard IHC practice). Purkinje-cell lipofuscin may complicate fluorescence interpretation; assess autofluorescence with an unstained section if using IF (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 chromogenic CD83 staining in paraffin sections by checking retrieval, compartment, cell identity and controls before comparing staining scores.
What retrieval conditions should I try first for CD83 staining in paraffin sections?
Use citrate pH 6.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval specification). Cool sections gradually, rinse consistently, and compare a known positive section in the same staining run; CD83 staining has been reported in lung macrophages and appendix germinal center cells (HPA tissue IHC). If signal is weak, test a different retrieval condition on serial sections while keeping antibody concentration and detection constant (standard IHC practice). Excess retrieval can damage section morphology or increase diffuse staining, so judge signal alongside tissue preservation (standard IHC practice). Record the condition used for every scored slide (standard IHC practice).
Can the CD83 record tell me how fixation will affect staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; the catalog image identifies paraffin-embedded human lung carcinoma tissue but does not state its fixative (A01777 caption). Do not infer fixation tolerance from the reported CD83 topology or glycosylation sites (UniProt Q01151 topology and glycosylation). When comparing specimens, document the fixative, fixation duration, processing history and section age, then stain comparable sections together (standard IHC practice). If staining differs across batches, compare morphology and a shared positive control before changing retrieval or antibody concentration (standard IHC practice). Treat any fixation effect observed in that comparison as specific to the tested material and workflow (standard IHC practice).
How should I assess membrane and cytoplasmic CD83 staining?
CD83 has an extracellular region at residues 20–144, a transmembrane segment at 145–166, and a cytoplasmic tail at 167–205 (UniProt Q01151 topology). Plasma membrane localisation is approved in the subcellular dataset, while Golgi localisation is also reported (HPA subcellular). Tissue IHC shows cytoplasmic staining in subsets of immune cells, including macrophages in lung (HPA tissue IHC). Score membrane-associated and cytoplasmic chromogen separately when morphology permits, and identify the stained cell population before assigning a result (standard IHC practice). Diffuse staining outside intact cells warrants comparison with a negative control and adjacent well-preserved tissue (standard IHC practice).
How can epitope position explain discordant CD83 staining?
CD83 has no isoforms listed in the supplied record, so an isoform-specific explanation is unsupported here (UniProt Q01151 isoforms). Its extracellular region spans residues 20–144, its cytoplasmic tail spans 167–205, and glycosylation sites are listed at 79, 96 and 117 (UniProt Q01151 topology and glycosylation). Check the antibody's documented immunogen or mapped epitope before interpreting discordant staining; no epitope position is supplied for A01777 (A01777 caption). Compare antibodies against distinct documented regions on serial sections with matched controls if available (standard IHC practice). Retrieval-dependent differences alone cannot establish which CD83 region an antibody recognizes (standard IHC practice).
How should I investigate CD83 by multiplex IF after chromogenic IHC?
For multiplex IF, pair CD83 with a validated marker that identifies the cell population being examined; activated dendritic cells and lymphocytes are among the reported CD83-expressing cells (UniProt Q01151 tissue specificity). Choose fluorophores after inspecting unstained tissue for autofluorescence, and place a weak CD83 channel away from the strongest background when possible (standard IF practice). Check whether the antibody recognizes the extracellular region at 20–144 or cytoplasmic tail at 167–205 before setting permeabilisation conditions (UniProt Q01151 topology). Intracellular epitopes require permeabilisation; excessive permeabilisation can compromise membrane patterns (standard IF practice). Validate each single-color stain and its negative controls before interpreting colocalisation (standard IF practice).
What should I check when CD83 chromogen appears widespread?
First inspect a no-primary control, reagent-only areas and tissue edges under the same development conditions (standard IHC practice). In enzyme-based chromogenic detection, check the peroxidase block and shorten chromogen development if background also appears without primary antibody (standard IHC practice). Increase blocking or washing only one variable at a time, and compare serial sections at the same exposure to detection reagents (standard IHC practice). CD83 tissue IHC has reported presumed off-target binding, so widespread staining deserves additional scrutiny (HPA tissue IHC reliability description). Keep intact-cell morphology and the expected immune-cell distribution in view when deciding whether faint staining is interpretable (HPA tissue IHC; standard IHC practice).
How should I quantify CD83-positive cells across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, because reported CD83 staining includes subsets of immune cells rather than uniform staining throughout tissue (HPA tissue IHC). Report the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories applied consistently across sections (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable tissue and state whether the denominator includes all nucleated cells or a defined cell population (standard IHC practice). Score membrane and cytoplasmic patterns separately when resolvable (UniProt Q01151 topology; HPA tissue IHC). Exclude folds, necrosis and poorly preserved edges using rules set before scoring (standard IHC practice).
How can I distinguish convincing CD83 staining from artefact?
Convincing staining should occur in intact cells with interpretable morphology and a plausible distribution; activated dendritic cells and lymphocytes express CD83, and lung macrophages show tissue staining (UniProt Q01151 tissue specificity; HPA tissue IHC). Membrane-associated staining fits CD83 topology, while cytoplasmic staining has also been reported in tissue IHC (UniProt Q01151 topology; HPA tissue IHC). Treat isolated edge staining, necrotic deposits and signal in the no-primary control as potential artefacts (standard IHC practice). Check endogenous enzyme activity if chromogen develops in controls without primary antibody (standard IHC practice). HPA notes presumed off-target binding, so compartment and cell identity should support any categorical positive call (HPA tissue IHC reliability description).
Boster reagents
Best CD83 / CD83 antigen IHC Antibodies
A01777 has an IHC image from paraffin-embedded human lung carcinoma tissue (catalog image caption) and lists human, mouse and rat reactivity (catalog: reactivity).
Real IHC data
Anti-CD83 (P130) Antibody
Cat # A01777
A01777 is listed for IHC in human, mouse and rat (catalog: applications; reactivity). Its IHC figure shows paraffin-embedded human lung carcinoma tissue (catalog image caption); no IF figure is supplied (catalog: IF image alts).
Which to pick: For tissue IHC, choose A01777; its own image shows paraffin-embedded human lung carcinoma tissue, but the fixative is unreported (catalog image caption). No listed SKU is validated for IF/ICC (catalog: applications; IF image alts). For cross-species IHC planning, A01777 lists human, mouse and rat reactivity, although its IHC image shows human tissue only (catalog: reactivity; image caption).
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 Cytoplasmic staining 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 conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining was observed and disregarded (HPA tissue IHC); Isoform / epitope: No isoforms; epitope side matters: residues 20–144 vs 167–205 (UniProt); Regulation: Expressed by activated lymphocytes and dendritic 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, PMC10868929. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CD83/immunohistochemistry-ihc-guide.