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- Table of Contents
CD83 / CD83 antigen · Western blot design guide
Design a Western Blot for CD83
Real validated CD83 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CD83 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
CD83 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~23 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated CD83 Western Blot Protocols
The M01777-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MOLT4 cell lysate (catalog M01777-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M01777-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected CD83 Western Blot Band Size?
CD83 has a predicted 23 kDa precursor; signal peptide cleavage, N-linked glycosylation, and oligomerization could affect migration, but no apparent band size is demonstrated.
| Band near 23 kDa | consistent with the predicted precursor mass, pending identity controls |
| Band below 23 kDa | may reflect removal of the 1–19 signal peptide |
| Band above 23 kDa | may reflect N-linked glycosylation at Asn79, Asn96, or Asn117 |
| Band near twice the monomer size under nonreducing conditions | may reflect retained CD83 homodimers; the annotated disulfide is intrachain |
💡Expected CD83 appearanceCD83's predicted precursor mass is 23 kDa; signal peptide cleavage, N-linked glycosylation, and oligomerization may affect migration, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted precursor mass | provides a 23 kDa sequence-based reference, not a validated apparent band size |
| N-linked glycosylation at Asn79 | may increase apparent mass if occupied |
| N-linked glycosylation at Asn96 | may increase apparent mass if occupied |
| N-linked glycosylation at Asn117 | may increase apparent mass if occupied |
| Signal peptide at residues 1–19 | cleavage makes mature CD83 smaller than its precursor |
| CD83 homodimer formation | could yield a band near twice the monomer size if the complex survives sample preparation |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | N-linked glycosylation or retained CD83 oligomers may increase apparent size | compare deglycosylated and reducing samples with a CD83 identity control |
| Band lower than expected | signal peptide cleavage may lower mass relative to the precursor | compare with a CD83-positive control and check antibody recognition of the detected form |
| Broad smear instead of sharp band | variable occupancy or composition at the three N-linked glycosylation sites is possible | compare treated and untreated samples after N-glycan removal |
| Multiple bands | glycosylation, signal peptide processing, or retained oligomers may contribute | compare reducing and deglycosylated samples with a CD83-positive control |
| Weak or no signal | CD83 is a single-pass membrane protein, so recovery may depend on membrane extraction | check membrane protein recovery and compare with a CD83-positive lysate |
Experiment evidence
Western blot images showing CD83 band
M01777-1 · CD83
A01777-1 · CD83
A01777 · CD83
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CD83 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CD83 in Western blot, you can use cerebellum lysate, the highest-scoring HPA positive tissue.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a membrane protein, CD83 may be underrepresented in soluble lysate fractions.
HPA tissue expression evidence for CD83
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for CD83, answered from its protein features.
How should CD83 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could an alternative CD83 isoform explain an extra band?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. An extra band cannot be assigned to a documented alternative isoform on this evidence; check its identity before labeling it CD83.
Which CD83 glycosylation sites matter when comparing bands?
PTM · UniProt lists N-linked sites at Asn79, Asn96, and Asn117, using full-length sequence numbering. Record that numbering when comparing antibody or paper descriptions, and do not assign a specific band to any one site from the annotation alone.
Does this guide establish induction of CD83?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for CD83?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01777-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How can CD83 bands be quantified consistently?
Quantitation · CD83 is annotated as a single-pass type I membrane protein with three N-linked glycosylation sites. Quantify the same defined band or band region across samples under consistent preparation conditions; establish band identity before combining signals at different apparent masses.
Why might CD83 migrate differently from its predicted 23 kDa mass?
Interpretation · The 23 kDa value is predicted. UniProt annotates a signal peptide at residues 1–19 and N-linked glycosylation sites at Asn79, Asn96, and Asn117. These features matter when interpreting apparent mass, but they do not establish a visible shift or explain a particular band without an observed mass.
UniProt describes CD83 as a monomer, homodimer, and homotrimer. Consider these when assessing higher bands, but oligomer annotation alone does not establish that those forms survive Western-blot preparation or identify an observed band.
UniProt lists one disulfide bond. Keep reduction conditions consistent when comparing samples, and do not attribute a changed band to that bond without supporting evidence.
UniProt annotates a signal peptide at residues 1–19 of the 205-residue sequence. Note whether a mass estimate refers to the full-length sequence or a processed protein. The annotation alone does not establish the apparent Western-blot mass.
UniProt describes CD83 as a monomer, homodimer, and homotrimer. Consider these when assessing higher bands, but oligomer annotation alone does not establish that those forms survive Western-blot preparation or identify an observed band.
UniProt lists one disulfide bond. Keep reduction conditions consistent when comparing samples, and do not attribute a changed band to that bond without supporting evidence.
UniProt annotates a signal peptide at residues 1–19 of the 205-residue sequence. Note whether a mass estimate refers to the full-length sequence or a processed protein. The annotation alone does not establish the apparent Western-blot mass.
Boster reagents
CD83 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-CD83 Rabbit Monoclonal Antibody
Cat # M01777-1
Real WB data
Anti-CD83 Antibody Picoband®
Cat # A01777-1
Real WB data
Anti-CD83 (P130) Antibody
Cat # A01777
Three anti-CD83 antibodies are listed. M01777-1 has a CD83 Western blot image in MOLT4 lysate; A01777 shows CD83 blots in HeLa, sp2/0, and PC12 lysates. Although A01777-1 has an image, its caption describes ADAM22, so it does not establish CD83 blot performance.
Which to pick: For MOLT4 lysate, consider M01777-1. For HeLa, sp2/0, or PC12 lysate, consider A01777, shown at 1:500. Both list human, mouse, and rat reactivity; the pictured samples are the documented contexts. A01777-1 lists human reactivity, but its image caption does not document CD83.
Source: BosterBio CD83 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q01151.
- Human Protein Atlas. CD83 tissue expression.
- PMC4300110 — target-verified WB comparison
- PMC4487981 — target-verified WB comparison
- PMC1940313 — target-verified WB comparison