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CD38 / ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 · IHC design guide
Design Immunohistochemistry for CD38
Plan CD38 chromogenic IHC in paraffin sections with lymph node or tonsil as high-staining controls (HPA tissue IHC). Assess membranous and cytoplasmic staining in lymphoid cells (HPA tissue IHC), accounting for CD38-positive immune cells when scoring (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CD38 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 | Membranous and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Distinct membrane and cytoplasm staining in lymphoid cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00193-3) | |
| Positive control | Lymph node+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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | CD38-positive immune cells can complicate cell-type scoring (HPA tissue IHC) | |
| Regulation | Lymphoid tissue enhanced (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; extracellular versus cytoplasmic epitope impact is unknown (UniProt) |
Section 1
Recommended CD38 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A00193-3) with two published CD38 chromogenic IHC protocols (PMC13357626; PMC6281009).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A00193-3) |
| Fixation | Image fixative and duration unreported (datasheet A00193-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A00193-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00193-3) |
| Primary antibody | Rabbit anti-CD38, 2μg/ml (datasheet A00193-3) |
| Primary incubation | Overnight at 4 °C (datasheet A00193-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00193-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CD38-positive staining in non-germinal center cells of lymph node (HPA tissue IHC: High). HPA tissue profile: Distinct cytoplasmic and membranous expression in hematopoietic and lymphoid tissue with immune cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A00193-3); neither published excerpt specifies retrieval (PMC13357626; PMC6281009).
Section 2
What Is the Expected CD38 Staining Pattern?
CD38 is a cell surface protein with one transmembrane segment and a predominantly extracellular domain (UniProt P28907 topology). In tissue IHC, expect membranous and cytoplasmic staining, especially in hematopoietic and lymphoid tissue (HPA tissue IHC). HPA rates the tissue staining profile Enhanced, citing high consistency between antibody staining and RNA expression (HPA tissue IHC). Interpret the signal by both compartment and cell type.
| Membranous staining with some cytoplasmic signal in lymph node or tonsil non-germinal center cells. | This fits the reported high staining in those cells and the broader membranous and cytoplasmic tissue pattern (HPA tissue IHC). Score the relevant cells rather than treating every unstained cell in the section as a failed result. |
| Predominantly nuclear staining, with little plausible membrane or cytoplasmic signal. | Treat this as an unexpected compartment pattern: CD38 is assigned to the cell surface and membrane (UniProt P28907 subcellular location), while HPA reports membranous and cytoplasmic tissue staining (HPA tissue IHC). Review controls and antibody specificity before calling it CD38. |
| Strong staining in respiratory epithelial cells of bronchus. | HPA reports CD38 as not detected in that cell population (HPA tissue IHC). Check whether the color follows the intended cells; cross-reactivity or endogenous chromogenic activity is possible (general IHC practice). A positive immune cell nearby would require separate scoring. |
| Color spreads across stroma, empty spaces, and many unrelated cell types. | A diffuse, poorly cell-associated deposit does not match the reported cell-specific pattern (HPA tissue IHC). Excess detection background or inadequate blocking can obscure true staining (general IHC practice); judge the section against its negative control. |
| No convincing staining in lymph node non-germinal center cells or tonsil non-germinal center cells. | Both populations are reported as high (HPA tissue IHC). Review the control section, retrieval, primary antibody dilution, detection reagents, and counterstain balance (general IHC practice) before interpreting an experimental negative. |
💡Expected CD38 appearanceCall a result positive when the relevant cells show clear membrane-associated staining, with possible cytoplasmic staining, especially in high-staining lymph node or tonsil non-germinal center cells (HPA tissue IHC; UniProt P28907 topology); isolated nuclear color or diffuse color across unrelated structures is suspect (HPA tissue IHC; general IHC practice).
| Cell population and tissue context | HPA reports high staining in lymph node and tonsil non-germinal center cells and in prostate and seminal vesicle glandular cells; bone marrow hematopoietic cells and spleen red-pulp cells are medium (HPA tissue IHC). Do not assume all glandular cells are negative. |
| Expected compartment | Residues 22–42 form the transmembrane segment; residues 43–300 are extracellular (UniProt P28907 topology). HPA nevertheless describes both membranous and cytoplasmic staining in tissue IHC (HPA tissue IHC), so cytoplasmic color alone needs cell-context and control review. |
| Negative cell controls | Adipose-tissue adipocytes and bronchial respiratory epithelial cells are reported as not detected (HPA tissue IHC). Use the named cell populations as comparison points; the HPA entry does not establish that every cell in either tissue is negative. |
| Evidence and antibody choice | The HPA tissue profile is Enhanced, and four listed antibodies have Enhanced IHC validation: HPA022132, HPA052381, CAB002493, and CAB025255 (HPA tissue IHC; HPA antibodies). This supports the reported pattern, without establishing identical staining in every specimen. |
| IF/ICC Q&A: Where should CD38 appear? | At the plasma membrane, the supported main location in HPA ICC-IF (HPA subcellular). HPA022132 has Supported ICC validation (HPA antibodies). Use the separate IF/ICC guide for that application; these observations do not define an IHC retrieval or dilution setting. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive lymphoid cells are blank. | An assay failure is possible because lymph node and tonsil non-germinal center cells are high in the reference profile (HPA tissue IHC). | Confirm that the positive control stained, then review retrieval, dilution, detection, and counterstain using the assay's validated conditions (general IHC practice). |
| Nuclear color dominates the section. | That distribution conflicts with the reported cell surface location and tissue membrane/cytoplasm pattern (UniProt P28907; HPA tissue IHC). | Compare the negative control and test a separately validated antibody or detection condition before assigning the signal to CD38 (general IHC practice). |
| Bronchial respiratory epithelium stains strongly. | That cell population is reported as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity could contribute (general IHC practice). | Inspect cell boundaries and the negative control; address endogenous chromogenic activity according to the detection system's standard controls (general IHC practice). |
| Prostate or seminal vesicle glands stain despite an expectation of immune-only signal. | Glandular cells in both tissues are reported high (HPA tissue IHC). | Score the glandular compartment separately from immune cells and compare the observed compartment with the HPA membranous and cytoplasmic pattern (HPA tissue IHC). |
| Background color obscures cell boundaries. | Nonspecific detection or insufficient blocking may produce diffuse color (general IHC practice). | Check the negative control, blocking, wash steps, and detection exposure before scoring cell-associated signal (general IHC practice). |
| Cytoplasmic staining appears without a clear membrane rim. | Cytoplasmic staining is included in the HPA tissue profile, although UniProt places CD38 at the cell surface (HPA tissue IHC; UniProt P28907). | Check whether staining occurs in an HPA-positive cell population and whether controls are clean; record the compartment explicitly rather than forcing a membrane-only score (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CD38 staining
A00193-3 · IHCIHC analysis of CD38 using anti-CD38 antibody (A00193-3). CD38 was detected in paraffin-embedded section of human tonsil tissue. Heat mediat
A00193-3 · IHCIHC analysis of CD38 using anti-CD38 antibody (A00193-3). CD38 was detected in paraffin-embedded section of human appendicitis tissue. Heat
A00193-3 · IFIF analysis of CD38 using anti-CD38 antibody (A00193-3). CD38 was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
M00193 · IHCIHC analysis of CD38 using anti-CD38 antibody (M00193). CD38 was detected in a paraffin-embedded section of human appendix tissue. Heat medi
M00193 · IHCIHC analysis of CD38 using anti-CD38 antibody (M00193). CD38 was detected in a paraffin-embedded section of human appendix tissue. Heat medi
M00193-7 · IHCHuman spleen was stained with anti-CD38 rabbit antibody
M00193-7 · IHCMouse spleen was stained with anti-CD38 rabbit antibody
M00193-8 · IHCRat pancreas was stained with anti-CD38 rabbit antibody
M00193-8 · IHCMouse pancreas was stained with anti-CD38 rabbit antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000004468-CD38/tissue/cerebral+cortex#imid_6084408
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000004468-CD38/tissue/colon#imid_6084350
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000004468-CD38/tissue/liver#imid_6084343
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000004468-CD38/tissue/kidney#imid_6084336
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA022132; green = CD38, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA022132; green = CD38, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CD38 IHC & IF
🧪Run tonsil first: non-germinal center cells should stain (HPA: High in tonsil non-germinal center cells; A00193-3 tissue-IHC caption: tonsil). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes). On the tonsil slide, assess morphologically distinct unstained cells as internal background; the HPA row does not establish which other cells must be negative (HPA: High in tonsil non-germinal center cells).
Positive control tissue: Lymph node (Non-germinal center cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CD38 in A-431, A-549, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a concentration-matched rabbit IgG isotype control for the rabbit primary, and CD38-knockout material as a biological negative (A00193-3 tissue-IHC caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and check endogenous biotin background in the biotin/SABC-DAB workflow, particularly on the tonsil slide (A00193-3 tissue-IHC caption: biotinylated secondary and SABC-DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A00193-3 paraffin-section caption leaves the fixative unreported (A00193-3 tissue-IHC caption: fixative not stated). That caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish that retrieval is required (A00193-3 tissue-IHC caption). Whether frozen sections or IF/ICC are easier is unreported; for this tonsil IHC workflow, endogenous peroxidase or biotin can confound DAB signal (A00193-3 tissue-IHC caption: biotin/SABC-DAB; standard IHC practice).
HPA tissue IHC evidence for CD38
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Prostate | Glandular cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | 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 → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CD38 IHC Tips
Troubleshoot CD38 chromogenic IHC in paraffin sections using the catalog antibody’s tonsil example and independent localisation evidence.
How should I adjust retrieval when CD38 staining is weak or diffuse?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections stained with A00193-3 (datasheet A00193-3). Use the reported tonsil workflow as a reference: it applied 2 μg/mL primary antibody overnight at 4°C, followed by biotinylated secondary antibody and DAB detection (datasheet A00193-3). If signal is weak, compare heating durations on adjacent sections while keeping antibody concentration, detection chemistry, and imaging conditions fixed (standard IHC practice). Favor the condition that improves staining in tonsil non-germinal center cells without increasing edge staining or negative-control signal (HPA: high in tonsil non-germinal center cells; standard IHC practice).
Can the available evidence tell me which fixation conditions preserve CD38 staining?
The selected paraffin tonsil caption does not state a fixative, so CD38 sensitivity to any particular fixation condition is unknown for A00193-3 (datasheet A00193-3). Record the actual fixative, fixation duration, processing history, and section age for each specimen before comparing staining intensity across cases (standard IHC practice). If fixation varies, compare similarly processed sections and include a consistently processed positive control in each staining run (standard IHC practice). Do not infer a preferred fixative from the observed tonsil staining, HPA tissue distribution, membrane topology, or glycosylation sites; none supplies a fixation comparison for this antibody (datasheet A00193-3; HPA tissue IHC; UniProt P28907).
Should CD38 appear only at the cell surface in paraffin sections?
Expect membrane-associated staining because CD38 is a cell-surface protein with a transmembrane segment at residues 22–42 and an extracellular region at 43–300 (UniProt P28907 topology). HPA also reports distinct membranous and cytoplasmic staining in hematopoietic and lymphoid tissue, so cytoplasmic signal alone is not sufficient grounds to discard a case (HPA tissue IHC). Inspect whether staining outlines cells or follows plausible intracellular profiles, and compare those patterns with adjacent negative-control sections before scoring (standard IHC practice). Diffuse staining across tissue edges, empty spaces, and unrelated cell populations deserves investigation before it is attributed to CD38 (standard IHC practice).
Can isoforms or glycosylation explain inconsistent CD38 staining?
CD38 has 2 annotated isoforms and 4 glycosylation sites, at residues 100, 164, 209, and 219 (UniProt P28907). Its short cytoplasmic region spans residues 1–21, whereas residues 43–300 are extracellular, making epitope location relevant to section staining and antibody comparisons (UniProt P28907 topology). The supplied evidence does not identify the A00193-3 epitope or establish isoform-specific recognition, so do not assign discordant staining to a particular isoform or glycan (datasheet A00193-3; UniProt P28907). When comparing antibodies, document each disclosed immunogen or epitope, apply the same retrieval and controls, and report unresolved differences as assay dependent (standard IHC practice).
How can IF help resolve ambiguous CD38 IHC cell identity?
Use a separate IF assay to compare CD38 with a validated marker for the suspected cell population; the IHC tissue profile includes high staining in tonsil non-germinal center cells (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence and spectral overlap, then include single-color and no-primary controls when interpreting colocalisation (standard IF practice). For an antibody against the extracellular region, test whether permeabilisation is necessary; an epitope in residues 1–21 would require intracellular access in intact cells (UniProt P28907 topology; standard IF practice). The supplied A00193-3 tonsil conditions describe chromogenic paraffin-section IHC and do not establish IF fixation or permeabilisation settings for that antibody (datasheet A00193-3).
How do I distinguish CD38 staining from chromogenic background?
First inspect a no-primary section processed through the same secondary antibody, detection complex, and DAB steps to locate reagent-dependent staining (standard IHC practice). The reported A00193-3 workflow used 10% goat serum, a biotinylated goat anti-rabbit secondary antibody, and a streptavidin-biotin detection complex (datasheet A00193-3). Include a peroxidase block in the chromogenic workflow and investigate endogenous biotin if the biotin-based detection system produces staining in no-primary controls (standard IHC practice). If background persists, optimize blocking, washes, primary concentration, and DAB development one variable at a time while monitoring tonsil non-germinal center cells as a positive reference (HPA tissue IHC; standard IHC practice).
What should I measure when comparing CD38 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue compartment before scoring, because CD38 staining may be membranous or cytoplasmic in hematopoietic and lymphoid tissue (HPA tissue IHC). For comparable cell populations, report the percentage of positive cells and an intensity-weighted H-score from 0–300, with staining thresholds set using controls (standard IHC practice). When cell abundance itself is the question, report CD38-positive cells per mm² of viable tissue and document excluded necrotic or damaged areas (standard IHC practice). Normalize comparisons to the number of eligible cells or viable tissue area, as appropriate, and keep retrieval, detection, imaging, and scoring rules consistent across sections (standard IHC practice).
What evidence supports a true CD38-positive IHC result?
Look for reproducible staining in an appropriate cell population with membrane-associated signal, allowing for the cytoplasmic pattern also reported in hematopoietic and lymphoid tissue (UniProt P28907 subcellular location; HPA tissue IHC). Tonsil non-germinal center cells provide a high-staining reference, while adipocytes are reported as undetected and can inform interpretation within an appropriate specimen (HPA tissue IHC). Treat staining confined to section edges, necrotic regions, or no-primary controls as suspect, especially with DAB and a biotin-based detection system (datasheet A00193-3; standard IHC practice). Confirm a disputed pattern using consistent processing, appropriate negative controls, and cell identity rather than intensity alone (standard IHC practice).
Boster reagents
Best CD38 / ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 IHC Antibodies
The catalog shows CD38 IHC in human tonsil and appendix and rat pancreas (IHC image captions), plus IF in A431 cells (A00193-3 IF caption).
Real IHC data
Anti-CD38 Antibody ®
Cat # A00193-3
Real IHC data
Anti-CD38 Rabbit Monoclonal Antibody
Cat # M00193
Real IHC data
Anti-CD38 Rabbit Monoclonal Antibody
Cat # M00193-7
Real IHC data
Anti-CD38 Rabbit Monoclonal Antibody
Cat # M00193-8
A00193-3 has human tonsil IHC and A431 cell IF images (A00193-3 image captions); M00193 has human appendix IHC images (M00193 IHC captions). M00193-7 has a human spleen IHC image and lists human reactivity (M00193-7 image caption and catalog); M00193-8 has a rat pancreas IHC image and lists human, mouse and rat reactivity (M00193-8 image caption and catalog).
Which to pick: For human tissue IHC, choose M00193: it is a rabbit monoclonal antibody with human IHC listed (M00193 catalog), and its appendix image uses paraffin sections with EDTA pH 8 retrieval; the fixative is unreported (M00193 IHC caption). For IF/ICC, choose A00193-3, which lists both applications and has an A431 cell IF image (A00193-3 catalog and IF caption). For cross-species IHC, choose M00193-8: it lists human, mouse and rat reactivity and shows rat and mouse pancreas IHC; retrieval and fixative are unreported in those images (M00193-8 catalog and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P28907 (CD38_HUMAN, ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1).
- Human Protein Atlas. CD38 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CD38 subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. CD38 antibody validation summary (4 antibodies).
- L-carnitine protects against methotrexate-induced hepatotoxicity via regulation of apoptosis and metabolic pathways. Naunyn-Schmiedeberg's archives of pharmacology 2026 — PMC13357626.
- The effect of chronic endometritis and treatment on patients with unexplained infertility. BMC women's health 2023 — PMC10311816.
- Prognostic Impact of CD38- and IgκC-Positive Tumor-Infiltrating Plasma Cells in Triple-Negative Breast Cancer. International journal of molecular sciences 2023 — PMC10607675.
- Inflammatory cell expression of Toll-like receptor-2 (TLR2) within refractory periapical granuloma. F1000Research 2018 — PMC6281009.
- PubMed PMID:2319135 — UniProt-cited evidence.
- PubMed PMID:9074508 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CD38 (ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1). Expected localisation Membranous and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00193-3). Validated IHC antibody: A00193-3 at 2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lymph node (Non-germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A00193-3); 2. Fixation: Image fixative and duration unreported (datasheet A00193-3); 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 retrieval: EDTA pH 8.0 (datasheet A00193-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00193-3); 8. Primary antibody: Rabbit anti-CD38, 2μg/ml (datasheet A00193-3); 9. Primary incubation: Overnight at 4 °C (datasheet A00193-3). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: CD38-positive immune cells can complicate cell-type scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms; extracellular versus cytoplasmic epitope impact is unknown (UniProt); Regulation: Lymphoid tissue enhanced (HPA tissue IHC). Evidence: UniProt P28907, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane., Datasheet A00193-3, PMC13357626, PMC6281009. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CD38/immunohistochemistry-ihc-guide.