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- Table of Contents
Plan chromogenic CD38 IHC in paraffin sections using the observed immune cell staining pattern and the protein’s membrane topology (HPA tissue IHC; UniProt). Use lymph node or tonsil as a high staining reference and assess membranous and cytoplasmic signal by cell population (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Membranous and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Membranous and cytoplasmic staining in immune 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 |
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00193-3) | |
| Caveat | Immune cells may complicate scoring in mixed tissues (HPA tissue IHC) | |
| Regulation | Induction not established in supplied evidence (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified; check membrane side (UniProt) |
The catalog antibody’s IHC-P protocol is followed by 4 published CD38 IHC protocols, covering liver, endometrium, breast tumors, and periapical lesions (PMC13357626; PMC10311816; PMC10607675; 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. |
CD38 is a cell-surface protein with residues 43–300 extracellular and residues 1–21 cytoplasmic (UniProt P28907 topology). In paraffin-section IHC, expect membranous and sometimes cytoplasmic staining in hematopoietic and lymphoid cells, with high staining in lymph-node and tonsil non-germinal-center cells and in prostate and seminal-vesicle glandular cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced, reflecting high agreement between staining and RNA expression (HPA tissue IHC).
| Distinct membrane staining in lymph-node or tonsil non-germinal-center cells, with some cytoplasmic signal. | This fits the cell-surface topology (UniProt P28907 topology) and the observed membranous and cytoplasmic lymphoid pattern (HPA tissue IHC). Judge signal by cell identity as well as compartment; adjacent cells need not stain equally. |
| High staining in prostate or seminal-vesicle glandular cells; medium staining in bone-marrow hematopoietic cells or splenic red-pulp cells. | These are documented cell-specific patterns, not evidence that every cell in each organ is positive (HPA tissue IHC). Compare the named cell population with its expected level before calling a section unusually weak or strong. |
| Predominantly nuclear staining, with no convincing membrane or cell-associated cytoplasmic pattern. | A nuclear-only pattern does not match the supported plasma-membrane location or the reported tissue IHC pattern (HPA subcellular ICC-IF; HPA tissue IHC). Treat it as suspect and check the matched negative control and detection background before scoring it as CD38. |
| Strong staining in adipocytes or bronchial respiratory epithelium, especially with little signal in expected positive cells. | Those specified cell populations were not detected by HPA tissue IHC (HPA tissue IHC). Unexpected signal can reflect cross-reactivity or detection background; assess its location and controls. A negative call for those cells does not make the whole tissue a negative control. |
| Widespread haze across cells and surrounding tissue, or no signal in lymph-node non-germinal-center cells. | Haze lacks the distinct cell-associated pattern reported for CD38, while absent signal in a documented high-staining population makes the run hard to interpret (HPA tissue IHC). Check controls and staining conditions before drawing a biological conclusion. |
| Topology and compartment | CD38 has one transmembrane segment at residues 22–42, an extracellular region at 43–300, and a cytoplasmic region at 1–21 (UniProt P28907 topology). The supported plasma-membrane location helps assess compartment; the record does not identify the catalog antibody's epitope (HPA subcellular ICC-IF). |
| Cell population and tissue | HPA reports high signal in specified lymphoid and glandular populations, medium signal in appendix lymphoid tissue, bone-marrow hematopoietic cells, and splenic red-pulp cells, and no detected signal in specified adipocytes and other listed cells (HPA tissue IHC). Use the named cells, not an organ-wide label, for interpretation. |
| Independent pattern support | HPA rates the tissue IHC pattern Enhanced and lists four antibodies with Enhanced IHC status (HPA tissue IHC; HPA antibodies). This supports the reported pattern but does not establish that an unlisted antibody, dilution, or retrieval condition will reproduce it. |
| Protein variants and modification | UniProt lists two isoforms and four glycosylation sites at residues 100, 164, 209, and 219 (UniProt P28907). These annotations alone do not predict isoform-specific staining, epitope accessibility, or a fixation effect; interpret the observed cell pattern against HPA tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in lymph-node or tonsil non-germinal-center cells | A documented high-staining population should provide an informative positive control (HPA tissue IHC); the failed run alone cannot identify the cause. | Check control tissue, antibody identity and working dilution, retrieval conditions, detection reagents, and counterstain visibility (general IHC practice). |
| Nuclear-only or sharply misplaced signal | The location conflicts with supported plasma-membrane localization and the reported tissue pattern (HPA subcellular ICC-IF; HPA tissue IHC). | Review cell boundaries and compare a matched negative control; repeat with an IHC-validated antibody if the pattern persists (general IHC practice; HPA antibodies). |
| Uniform chromogenic haze or granular staining across unrelated cells | Diffuse signal obscures the distinct cell-associated pattern reported by HPA (HPA tissue IHC); background can arise during chromogenic detection (general IHC practice). | Review blocking and washes; if using peroxidase detection, verify endogenous-peroxidase control and compare a no-primary control (general IHC practice). |
| Unexpected strong signal in adipocytes or bronchial respiratory epithelium | HPA did not detect CD38 in those specified cells (HPA tissue IHC); cross-reactivity or detection activity is possible, not proven. | Confirm cell identity and compartment, examine the negative control, and compare an independently validated IHC antibody where available (general IHC practice; HPA antibodies). |
| A negative field is interpreted as a negative organ | HPA reports staining for particular cell populations: for example, adipocytes were not detected, while immune cells occur in several tissues (HPA tissue IHC). | Score the named cell type and inspect a documented positive population on the same run before interpreting absence of signal (HPA tissue IHC; general IHC practice). |
| Can an ICC/IF image settle a disputed paraffin-section IHC result? | HPA supports plasma-membrane localization in ICC-IF and lists A-431 and A-549 images, while its tissue IHC record describes membranous and cytoplasmic staining (HPA subcellular ICC-IF; HPA tissue IHC). | Use ICC/IF as a compartment cross-check; judge the paraffin section against tissue IHC cell patterns and IHC controls (HPA tissue IHC; general IHC practice). |
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.
| 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 → |
| 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 → |
Use the paraffin-section tonsil image as the starting point for CD38 chromogenic IHC, then check cell identity, staining compartment, and controls (caption A00193-3; HPA tissue IHC).
The catalog includes anti-CD38 IHC images from human tonsil, appendix and spleen, plus mouse and rat pancreas (IHC captions); an IF image shows A431 cells (A00193-3 IF caption).
A00193-3 shows human tonsil IHC and A431 IF (A00193-3 image captions); M00193 shows human appendix IHC (M00193 IHC caption). M00193-7 shows human spleen and appendix IHC (M00193-7 IHC captions); M00193-8 shows rat and mouse pancreas IHC (M00193-8 IHC captions).
Which to pick: For human tissue IHC, M00193 is a rabbit monoclonal with an EDTA-retrieved paraffin appendix image; the fixative is unreported (catalog: M00193; M00193 IHC caption). For IF/ICC, A00193-3 lists both applications and has an A431 IF image; its tonsil IHC caption describes EDTA-retrieved paraffin sections but reports no fixative (catalog: A00193-3; A00193-3 image captions). For mouse or rat tissue IHC, M00193-8 is a rabbit monoclonal with both species listed and pancreas IHC images; those captions do not report processing or fixative (catalog: M00193-8; M00193-8 IHC captions).