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- Table of Contents
Plan paraffin-section CCND1 IHC around its general nuclear staining pattern (HPA tissue IHC). Use high-staining appendix lymphoid tissue and undetected prostate glandular cells as reference populations (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | General nuclear staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9370) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Prostate |
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9370) | |
| Caveat | Signal varies by cell type; prostate glands are undetected (HPA tissue IHC) | |
| Regulation | G1/S-linked expression (UniProt) | |
| Isoform / epitope | No isoforms or processing annotated (UniProt) |
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet: PB9370). Published IHC protocols below cover chondroid lipoma (PMC3087950), endometrial tissue (PMC11816803), and plasma cell myeloma (PMC6676137).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet PB9370) |
| Fixation | Image fixative and duration unreported (datasheet PB9370); 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 PB9370); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9370) |
| Primary antibody | Rabbit anti-CCND1, 2-5μg/ml (datasheet PB9370) |
| Primary incubation | Overnight at 4 °C (datasheet PB9370) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9370) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCND1-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
CCND1 is predominantly nuclear in tissue IHC (HPA: general nuclear expression; reliability Supported, with medium consistency between staining and RNA). UniProt also lists cytoplasm and nuclear membrane, but no transmembrane segment (UniProt P24385: subcellular location and topology). Expect staining in the cell populations HPA reports as positive, including appendix lymphoid tissue and breast myoepithelial cells (HPA: High in both).
| Distinct nuclear staining in appendix lymphoid tissue or breast myoepithelial cells. | This matches the reported cell populations and dominant compartment (HPA: High in both; general nuclear expression). Compare nuclei within the relevant population, since a positive tissue does not imply that every cell should stain (standard IHC interpretation). |
| Predominantly diffuse cytoplasmic or surface staining, with little nuclear signal. | Question whether this is the intended result: tissue IHC is generally nuclear (HPA: general nuclear expression). UniProt lists cytoplasm and nuclear membrane, so minor nonnuclear signal alone cannot establish an artefact; assess its distribution against nuclear staining and controls (UniProt P24385: subcellular location; standard IHC practice). |
| Strong staining mainly in a cell population other than the one expected for the selected tissue. | Check cell identity against the tissue-specific HPA entry; for example, the reported breast population is myoepithelial cells (HPA: High in breast myoepithelial cells). Unexpected signal can reflect antibody cross-reactivity or endogenous detection activity; omission and detection controls help distinguish them (standard IHC practice). |
| Uniform haze across cells and surrounding tissue, without clear nuclear boundaries. | This does not resemble HPA's general nuclear pattern (HPA: tissue IHC profile). Nonspecific antibody or detection background is possible; review blocking, reagent exposure and primary-omission controls before scoring nuclei (standard IHC practice). |
| No nuclear signal in a known-positive cell population. | First verify that the expected cells are present: HPA reports High staining in kidney glomerular cells and bone-marrow hematopoietic cells, among others (HPA: tissue IHC). If present, assess the antibody and detection controls and the established IHC-P workflow; an empty slide alone does not establish absent CCND1 (standard IHC practice). |
| Tissue and cell selection | HPA reports High staining in appendix lymphoid tissue, bone-marrow hematopoietic cells, breast myoepithelial cells, and several other specified populations; prostate glandular cells are Not detected (HPA: tissue IHC). Choose controls by cell population, and do not treat that prostate entry as a claim about every cell in the section (standard IHC interpretation). |
| Antibody validation | One listed antibody has Supported IHC status; the other has ICC Supported status without a listed IHC status (HPA: CAB080462 IHC Supported; HPA027802 ICC Supported). These application-specific labels do not make their results interchangeable (HPA: antibody validation; standard IHC interpretation). |
| Antigen retrieval | Use the retrieval conditions established for the IHC-validated antibody and compare control sections when optimizing (standard IHC practice). The supplied UniProt and HPA records do not report CCND1-specific retrieval requirements or fixation sensitivity. |
| Protein features | CCND1 is a 295-residue chain with no signal peptide, propeptide or transmembrane segment, and UniProt records phosphorylation at Thr286 (UniProt P24385: processing, topology and modified residue). None of these entries identifies this antibody's epitope or demonstrates that phosphorylation changes its IHC signal. |
| IF/ICC Q&A: Where should signal appear? | HPA places CCND1 in the nucleoplasm and lists BJ and U2OS ICC-IF images (HPA: supported nucleoplasmic location; ICC-IF image list). Use that as an IF localisation expectation; it is separate from the tissue IHC evidence (HPA: subcellular and tissue IHC records). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control section has no nuclear chromogen. | The expected cells may be absent from the viewed field, or the antibody, retrieval or detection workflow may have failed (HPA: cell-specific High entries; standard IHC practice). | Locate the HPA-listed population, then check the established IHC-P conditions and a working detection control; interpret the test section only after the positive control performs (standard IHC practice). |
| Prostate glandular cells stain strongly. | HPA reports this population as Not detected, making the result discordant with its tissue entry; nonspecific antibody or detection signal is possible (HPA: prostate glandular cells; standard IHC interpretation). | Confirm glandular cell identity and compare primary-omission and detection controls. Reassess antibody conditions if nuclear staining persists only with primary antibody (standard IHC practice). |
| Chromogen is widespread outside nuclei. | A broad nonnuclear pattern conflicts with HPA's general nuclear tissue profile; background or cross-reactivity may contribute (HPA: tissue IHC profile; standard IHC practice). | Compare omission controls, assess background in surrounding tissue, and optimize blocking or reagent exposure while retaining a nuclear positive control (standard IHC practice). |
| Staining appears only in unexpected cell types. | The cell population may have been misidentified, or signal may reflect cross-reactivity or endogenous detection activity (standard IHC interpretation). HPA levels apply to named cells within each tissue (HPA: tissue IHC entries). | Review tissue morphology and the corresponding HPA cell entry; use primary-omission and detection controls to test whether signal depends on the primary antibody (standard IHC practice). |
| Nuclear signal is weak but visible. | A low-staining population may have been chosen: HPA lists Low staining in thyroid and adrenal glandular cells, among others (HPA: tissue IHC). Weak signal by itself does not identify a technical failure. | Compare with a High HPA-listed population on a control section and score the selected cells against local background under the same detection conditions (HPA: tissue levels; standard IHC practice). |
| IF/ICC signal looks different from tissue IHC. | The evidence describes different preparations: nucleoplasmic localisation in ICC-IF and general nuclear expression in tissue IHC (HPA: subcellular and tissue IHC records). | Interpret each against its own HPA observation and application-specific antibody validation; do not use ICC Support as proof of IHC performance (HPA: subcellular location and antibody validation). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot CCND1 staining in paraffin sections by checking retrieval, nuclear localisation, controls and cell level scoring (datasheet PB9370; HPA: general nuclear expression).
Anti-CCND1 antibodies have IHC images from human tissues and mouse and rat samples (catalog IHC image captions); IF/ICC images are available from human cells (catalog IF image captions).
PB9370 shows IHC in paraffin sections of human colon and endometrial cancer and rat lung (PB9370 IHC captions); M00149-5 shows IHC staining in human esophagus and tonsil, rat spleen, and mouse colon (M00149-5 IHC captions). M00149-1 shows IHC in paraffin sections of human bladder, pituitary tumor, squamous cell carcinoma, and ovarian cancer, plus IF in MCF-7 cells (M00149-1 IHC and IF captions).
Which to pick: For paraffin-section IHC, start with PB9370: its human colon cancer image specifies EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (PB9370 IHC caption); the fixative is unreported (PB9370 IHC caption). For IF/ICC, choose rabbit monoclonal M00149-1, which lists both applications and has an IF image in MCF-7 cells (M00149-1 catalog applications and IF caption). For IHC across species, consider rabbit monoclonal M00149-5: its captions show human esophagus and tonsil, rat spleen, and mouse colon (M00149-5 catalog title and IHC captions); tissue processing and fixative are unreported in those captions (M00149-5 IHC captions).