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CEACAM1 / Cell adhesion molecule CEACAM1 · IHC design guide
Design Immunohistochemistry for CEACAM1
Plan CEACAM1 chromogenic IHC in paraffin sections using gut enterocyte microvilli as a positive staining pattern (HPA tissue IHC). Use the catalog antibody at 1:50 (datasheet M00923-2), and score membrane staining by cell type against negative controls (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CEACAM1 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 | Enterocyte microvilli and epithelial membranes (HPA tissue IHC) | |
| Staining pattern | Membranous staining, strongest at gut enterocyte microvilli (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00923-2) | |
| Positive control | Appendix+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 paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Staining may include products of other genes (HPA tissue IHC) | |
| Regulation | No staining-linked induction specified (UniProt) | |
| Isoform / epitope | 11 isoforms; check extracellular versus cytoplasmic epitope (UniProt) |
Section 1
Recommended CEACAM1 IHC & IF Protocols
The catalog antibody protocol is followed by published CEACAM1 IHC methods for lymphoma and myeloma tissue microarrays, lung cancer specimens, and liver graft sections (PMC13312280; PMC3728234; PMC6686173).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M00923-2) |
| Fixation | Image fixative and duration unreported (datasheet M00923-2); 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 M00923-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00923-2) |
| Primary antibody | Rabbit monoclonal (clone 19C63) anti-CEACAM1, 1:50 (datasheet M00923-2) |
| Primary incubation | Overnight at 4 °C (datasheet M00923-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00923-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CEACAM1-positive staining in enterocytes - Microvilli of appendix (HPA tissue IHC: High). HPA tissue profile: Membranous expression in a few tissues, most abundant in glandular epithelia of the gastrointestinal tract, bile canaliculi of liver and intercalated ducts of salivary gland. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: M00923-2); citrate pH 6.0 is a published alternative for lung cancer sections (PMC3728234).
Section 2
What Is the Expected CEACAM1 Staining Pattern?
CEACAM1 is predominantly at lateral cell membranes, with apical, basal, junctional and microvillus locations also reported; it has one transmembrane segment and an extracellular domain (UniProt P13688 topology and subcellular location). In tissue IHC, expect the clearest staining along gastrointestinal enterocyte microvilli (HPA: High in appendix, colon and rectum). HPA rates tissue staining reliability Enhanced, while noting medium consistency with RNA data and possible detection of proteins from more than one gene (HPA: tissue IHC reliability).
| High staining outlines enterocyte microvilli in appendix, colon or rectum (HPA: High in these cells). | This matches the strongest supplied tissue pattern. A crisp surface distribution is consistent with CEACAM1 membrane and microvillus localization; assess the stained structure as well as its intensity (HPA: tissue IHC; UniProt P13688 subcellular location). |
| Strong nuclear staining dominates, with little discernible epithelial surface staining. | Nuclear dominance does not match the reported CEACAM1 locations (UniProt P13688 subcellular location). Treat it as a possible staining artifact and reassess the slide and controls before calling it CEACAM1 positive (standard IHC interpretation). |
| Strong staining appears in cells reported as undetected, such as adipocytes in adipose tissue or respiratory epithelial cells in bronchus (HPA: Not detected in those cells). | This is discordant with the supplied tissue observations. Cross-reactivity or endogenous detection activity is possible; the HPA tissue summary also cautions that staining may detect proteins from more than one gene (HPA: tissue IHC reliability; standard IHC interpretation). |
| Broad, diffuse color covers tissue and background without a clear cell boundary or microvillus pattern. | That distribution is hard to reconcile with the reported membrane predominance (UniProt P13688 subcellular location). Review background and detection controls before scoring cells; diffuse color alone does not establish a positive CEACAM1 result (standard IHC interpretation). |
| No signal appears in appendix, colon or rectum enterocyte microvilli (HPA: High in those cells). | The expected positive structure is missing, so this run cannot support a negative interpretation on its own. Check tissue preservation, retrieval, reagent performance and detection controls using the validated IHC workflow (standard IHC practice). |
💡Expected CEACAM1 appearanceCall positive a distinct enterocyte microvillus or epithelial membrane pattern, strongest in appendix, colon or rectum; isolated nuclear or diffuse background color is suspect (HPA: tissue IHC; UniProt P13688 subcellular location).
| Membrane topology (UniProt P13688 topology) | CEACAM1 has an extracellular region at residues 35–428, one transmembrane segment at 429–452 and a cytoplasmic region at 453–526. Interpret staining against the expected membrane distribution; the supplied record does not locate the antibody epitope (UniProt P13688 topology). |
| Tissue and cell context (HPA: tissue IHC) | Appendix, colon and rectum enterocyte microvilli are High; duodenum and small intestine enterocyte microvilli are Medium (HPA: tissue IHC). A weaker result in a Medium site should be judged against that site's reference level, not the strongest gastrointestinal fields. |
| Restricted structures outside the intestine (HPA: tissue IHC) | The HPA profile describes bile canaliculi in liver and intercalated ducts in salivary gland, while listing hepatocytes and salivary gland glandular cells as Low (HPA: tissue IHC). Examine the named structures before interpreting a broadly stained liver or salivary section. |
| Antibody and profile confidence (HPA: tissue IHC and antibody validation) | HPA lists IHC Supported for HPA011041 and IHC Enhanced for CAB002146 (HPA: antibody validation). Its overall tissue reliability is Enhanced, with medium staining–RNA consistency and a caution about proteins from more than one gene; a plausible pattern still needs controls (HPA: tissue IHC reliability). |
| Isoforms and processing (UniProt P13688 isoforms and processing) | UniProt lists 11 isoforms, a signal peptide at residues 1–34, and a mature chain beginning at residue 35 (UniProt P13688). Without an epitope assignment, the supplied evidence cannot establish which isoforms the IHC-validated antibody detects. |
| Additional reported locations (UniProt P13688 subcellular location) | UniProt also reports secreted CEACAM1 and secretory-vesicle membrane localization (UniProt P13688 subcellular location). Such annotations broaden the biological context, but they do not make uniform cytoplasmic color a sufficient positive call when the tissue reference is membranous (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive gastrointestinal tissue is blank. | A technical run failure is possible when HPA-High enterocyte microvilli have no signal (HPA: appendix, colon and rectum; standard IHC practice). | Confirm the expected cells are present, then review the validated antigen-retrieval and detection steps and a run control (standard IHC practice). No target-specific fixation sensitivity or retrieval condition is established by the supplied sources. |
| Diffuse chromogen obscures cell borders. | Background can arise from detection chemistry, insufficient blocking or an overly strong reagent condition (standard IHC practice). | Inspect a no-primary control, review blocking and detection, and use the validated antibody conditions before scoring. Seek a resolved membrane or microvillus pattern (standard IHC practice; HPA: tissue IHC). |
| Nuclear staining is the dominant finding. | A nuclear pattern conflicts with the supplied membrane, junction, vesicle and secreted locations (UniProt P13688 subcellular location). | Check counterstain and detection controls, then compare the same run with HPA-High enterocyte microvilli. Do not score nuclei alone as CEACAM1 positive (standard IHC interpretation; HPA: tissue IHC). |
| A reportedly negative cell population stains strongly. | Off-target binding or endogenous detection activity may explain staining that conflicts with an HPA Not detected entry; HPA also cautions about detection of proteins from more than one gene (HPA: tissue IHC reliability; standard IHC practice). | Verify the cell identity and compare no-primary and known-positive controls. Interpret the discordant result cautiously, especially if the expected membrane pattern is absent (standard IHC interpretation; UniProt P13688 subcellular location). |
| Liver or salivary gland looks uniformly strong. | Uniform intensity exceeds the Low cell-level entries and loses the canalicular or duct emphasis in the HPA profile (HPA: liver and salivary gland tissue IHC). | Inspect bile canaliculi or intercalated ducts at higher magnification and compare background controls before assigning a tissue-wide score (HPA: tissue IHC; standard IHC interpretation). |
| IF/ICC question: should a bright intracellular signal be accepted as the expected pattern? | HPA gives a Membrane subcellular summary but supplies no ICC-IF image-bearing cell lines or detailed main location; UniProt also reports secretory-vesicle membrane localization (HPA: subcellular ICC-IF; UniProt P13688 subcellular location). | Use membrane localization as the main comparison and interpret intracellular signal with controls and the separate IF/ICC guide. The supplied HPA ICC-IF record does not establish a cell-line-specific pattern (HPA: subcellular ICC-IF; standard IF interpretation). |
Experiment evidence
IHC & IF images showing CEACAM1 staining
M00923-2 · IHCIHC analysis of CEACAM1 using anti-CEACAM1 antibody (M00923-2). CEACAM1 was detected in a paraffin-embedded section of human colorectal aden
M00923-2 · IHCIHC analysis of CEACAM1 using anti-CEACAM1 antibody (M00923-2). CEACAM1 was detected in a paraffin-embedded section of human liver cancer ti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000079385-CEACAM1/tissue/cerebral+cortex#imid_716384
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000079385-CEACAM1/tissue/colon#imid_716310
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000079385-CEACAM1/tissue/liver#imid_716319
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000079385-CEACAM1/tissue/kidney#imid_716320
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CEACAM1 IHC & IF
🧪Run colon first: enterocyte microvilli should stain strongly (HPA: High in colon enterocyte microvilli); use adipose tissue adipocytes as the separate negative comparator (HPA: Not detected in adipocytes). On the colon slide, assess morphologically identifiable non-enterocyte cells for background-only staining as provisional internal negatives; the supplied HPA row does not establish their CEACAM1 status (HPA: colon row specifies enterocyte microvilli only).
Positive control tissue: Appendix (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CEACAM1; derive a cell-line control from the positive tissue's cell type (Enterocytes - Microvilli) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control, alongside CEACAM1-knockout tissue or cells processed as a biological negative (selected-SKU caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase and check for residual DAB signal in blood or inflammatory cells on the colon section (selected-SKU caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). The reported IHC procedure uses heat-mediated EDTA retrieval at pH 8.0; this supports testing that retrieval condition but does not establish that retrieval is required (selected-SKU tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; in colon, distinguish apical microvillar staining from residual peroxidase-associated DAB signal (HPA: High in colon enterocyte microvilli; selected-SKU caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for CEACAM1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Appendix | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Rectum | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Duodenum | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | 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 CEACAM1 IHC Tips
Troubleshoot CEACAM1 staining in paraffin sections by checking retrieval, membrane localisation, antibody specificity and cell level scoring before interpreting chromogenic signal.
How should I optimise retrieval when CEACAM1 staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0, the condition used for the catalog antibody in a paraffin section (datasheet M00923-2). Keep section thickness, heating time and cooling conditions consistent across comparison slides, then assess whether membrane staining improves without increased tissue damage (standard IHC practice). The documented example used a 1:50 primary antibody dilution overnight at 4°C, so check those conditions before changing retrieval chemistry (datasheet M00923-2). If staining remains weak, compare a second retrieval condition on matched sections while retaining the EDTA condition as the reference (standard IHC practice).
Could fixation explain inconsistent CEACAM1 staining between paraffin blocks?
The selected paraffin section caption does not report a fixative, and target specific fixation sensitivity is unknown for this antibody (datasheet M00923-2). Record each block’s fixative, fixation duration and processing history before comparing staining, because those variables can affect antigen accessibility in paraffin IHC (standard IHC practice). Run blocks with different histories in the same staining batch, alongside a consistently processed reference section, to distinguish block effects from reagent or detection changes (standard IHC practice). Do not assign a CEACAM1 specific fixation threshold from its membrane topology or reported tissue distribution; neither establishes fixation tolerance (UniProt P13688 topology; HPA tissue IHC).
Which staining pattern supports CEACAM1 localisation in a tissue section?
Prioritise a defined cell surface pattern: CEACAM1 is reported at lateral, apical and basal membranes, junctions, microvilli and the hepatocyte canalicular domain (UniProt P13688 subcellular location). Predominantly lateral membrane localisation is reported, while HPA describes membranous staining most abundant in gastrointestinal glandular epithelia (UniProt P13688 subcellular location; HPA tissue IHC). Inspect the epithelial border at high magnification and compare it with a no primary control before accepting diffuse cytoplasmic DAB as specific staining (standard IHC practice). Secreted and secretory vesicle membrane locations are also recorded, so compartment alone cannot establish antibody specificity (UniProt P13688 subcellular location).
Can this IHC antibody distinguish CEACAM1 isoforms or a cytoplasmic epitope?
Do not infer isoform selectivity from a positive paraffin section: the supplied caption gives staining conditions but no mapped antibody epitope or isoform validation (datasheet M00923-2). CEACAM1 has 11 listed isoforms, with residues 35–428 extracellular and 453–526 cytoplasmic in the recorded topology (UniProt P13688 isoforms and topology). A cytoplasmic tail antibody could read differently from an extracellular domain antibody if their recognised sequences differ among isoforms; confirm the immunogen and tested isoforms before making that claim (UniProt P13688 isoforms and topology; standard antibody validation practice). Its extracellular region has 20 listed glycosylation sites, another reason to validate an antibody’s accessible epitope experimentally (UniProt P13688 glycosylation).
How can I adapt the CEACAM1 localisation check to multiplex IF?
Treat multiplex IF as a separate validation experiment: the supplied catalog example documents chromogenic staining of a paraffin section, while HPA lists no ICC/IF images here (datasheet M00923-2; HPA subcellular). Pair CEACAM1 with a marker that identifies the expected epithelial cells, and inspect whether signal follows their cell borders rather than unrelated autofluorescent structures (HPA tissue IHC; UniProt P13688 subcellular location; standard IF practice). Choose fluorophores after measuring tissue autofluorescence and include single stain controls for spectral overlap (standard IF practice). For an extracellular epitope, assess staining without permeabilisation first; for a cytoplasmic tail epitope, optimise permeabilisation after fixation and verify epitope identity (UniProt P13688 topology; standard IF practice).
How do I reduce diffuse DAB or stromal background without losing membrane signal?
The catalog example used 10% goat serum blocking, a 1:50 primary dilution overnight at 4°C, and peroxidase detection with DAB (datasheet M00923-2). First examine a no primary section and apply an endogenous peroxidase block as a general chromogenic IHC control, especially when signal appears outside cell borders (standard IHC practice). Titrate primary antibody and secondary detection on matched sections, preserving the documented condition as a reference and checking that expected membrane detail remains visible (datasheet M00923-2; standard IHC practice). HPA warns that its tissue staining can reflect proteins from more than one gene, so a clean negative control alone does not establish CEACAM1 specificity (HPA tissue IHC).
What scoring method best captures CEACAM1 membrane staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and membrane compartment before scoring, since HPA reports strong enterocyte microvillus staining in colon, rectum and appendix (HPA tissue IHC). For epithelial regions, record the percentage of positive eligible cells and membrane intensity, then calculate an H-score using the same intensity bins across slides (standard IHC scoring practice). Normalise counts to the number of evaluable cells or, when measuring stained area, to evaluable tissue area in mm²; exclude folds and necrosis by a prespecified rule (standard IHC quantification practice). Keep apical and lateral membrane scores separate when polarity matters, because both are documented CEACAM1 locations (UniProt P13688 subcellular location).
When should an apparent CEACAM1 positive cell be considered artefactual?
Give greater weight to crisp membrane signal in an anatomically plausible cell population: HPA reports high staining in appendix, colon and rectum enterocyte microvilli (HPA tissue IHC). Question isolated nuclear staining or diffuse deposits without a cell border, because the recorded CEACAM1 locations centre on membranes, junctions and secretory compartments (UniProt P13688 subcellular location; standard IHC interpretation). Check tissue edges, folds and necrotic regions against adjacent intact tissue, and review a no primary slide for endogenous enzyme or detection artefacts (standard IHC practice). HPA assigns Enhanced reliability yet cautions that staining may reflect proteins from more than one gene; independent specificity evidence is needed for a definitive CEACAM1 assignment (HPA tissue IHC).
Boster reagents
Best CEACAM1 / Cell adhesion molecule CEACAM1 IHC Antibodies
M00923-2 has IHC images from human paraffin sections (catalog IHC captions) and is listed for IF/ICC (catalog applications); no IF image is supplied (catalog IF images).
Real IHC data
Anti-CEACAM1 Rabbit Monoclonal Antibody
Cat # M00923-2
M00923-2 will render with its human colorectal adenocarcinoma paraffin-section IHC figure (catalog IHC caption). The catalog also shows IHC in human liver cancer paraffin sections and lists IF/ICC applications, without an IF image (catalog IHC captions; catalog applications; catalog IF images).
Which to pick: Choose M00923-2 for human paraffin-section IHC: its own figure uses EDTA retrieval at pH 8.0, antibody at 1:50, and DAB detection; the fixative is unreported (M00923-2 IHC caption). For IF/ICC, M00923-2 is listed at 1:50, though no IF image is supplied (catalog applications; catalog IF dilution; catalog IF images). No cross-species choice is supported because M00923-2 lists human reactivity only (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P13688 (CEAM1_HUMAN, Cell adhesion molecule CEACAM1).
- Human Protein Atlas. CEACAM1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CEACAM1 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. CEACAM1 antibody validation summary (2 antibodies).
- CEACAM1 expression by immunohistochemistry in B-cell lymphomas and plasma cell myeloma. American journal of clinical pathology 2026 — PMC13312280.
- Clinical and experimental studies regarding the expression and diagnostic value of carcinoembryonic antigen-related cell adhesion molecule 1 in non-small-cell lung cancer. BMC cancer 2013 — PMC3728234.
- Hydrogen Flush After Cold Storage as a New End-Ischemic Ex Vivo Treatment for Liver Grafts Against Ischemia/Reperfusion Injury. Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society 2018 — PMC6686173.
- The CEACAM1 expression is decreased in the liver of severely obese patients with or without diabetes. Diagnostic pathology 2011 — PMC3104481.
- PubMed PMID:2537311 — UniProt-cited evidence.
- PubMed PMID:2457922 — UniProt-cited evidence.
- PubMed PMID:2025273 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CEACAM1 (Cell adhesion molecule CEACAM1). Expected localisation Enterocyte microvilli and epithelial membranes (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00923-2). Validated IHC antibody: M00923-2 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M00923-2); 2. Fixation: Image fixative and duration unreported (datasheet M00923-2); 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 M00923-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00923-2); 8. Primary antibody: Rabbit monoclonal (clone 19C63) anti-CEACAM1, 1:50 (datasheet M00923-2); 9. Primary incubation: Overnight at 4 °C (datasheet M00923-2). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Staining may include products of other genes (HPA tissue IHC); Isoform / epitope: 11 isoforms; check extracellular versus cytoplasmic epitope (UniProt); Regulation: No staining-linked induction specified (UniProt). Evidence: UniProt P13688, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet M00923-2, PMC13312280, PMC3728234. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CEACAM1/immunohistochemistry-ihc-guide.