This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CEACAM1 / Cell adhesion molecule CEACAM1 · IHC design guide
Design Immunohistochemistry for CEACAM1
Plan CEACAM1 paraffin IHC using intestinal enterocyte microvilli as a positive control (HPA tissue IHC). Assess membranous staining with the catalog antibody at 1:50 (datasheet M00923-2), while accounting for possible staining of protein from more than one gene (HPA tissue IHC).
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 | Membranous staining in GI glands and bile canaliculi (HPA tissue IHC) | |
| Staining pattern | Membranous in GI glands, bile canaliculi and salivary ducts (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 fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may reflect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | 11 isoforms; check extracellular versus cytoplasmic epitopes (UniProt) |
Section 1
Recommended CEACAM1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet M00923-2) with 3 published CEACAM1 paraffin-section IHC workflows (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 for the catalog antibody (datasheet M00923-2); use citrate pH 6.0 when reproducing the lung study (PMC3728234).
Section 2
What Is the Expected CEACAM1 Staining Pattern?
CEACAM1 is a membrane protein with extracellular and cytoplasmic regions; UniProt places it predominantly on lateral membranes and also at apical membranes and junctions (UniProt P13688 topology and subcellular location). In IHC, expect staining in intestinal enterocytes, especially their microvilli, and at liver bile canaliculi (HPA tissue IHC). HPA rates the tissue profile Enhanced but reports only medium consistency with RNA and cautions that staining may target more than one gene (HPA tissue IHC).
| Crisp enterocyte border staining, strongest at microvilli in colon, rectum or appendix (HPA tissue IHC). | This fits the reported High enterocyte microvillus staining in those tissues (HPA tissue IHC) and the apical, lateral and microvillus membrane locations assigned to CEACAM1 (UniProt P13688 subcellular location). Assess the cell border rather than counting all brown material as specific signal (general IHC practice). |
| Predominantly nuclear or broad, uniform cytoplasmic staining with little membrane definition. | That distribution is atypical for the predominant membrane and junctional pattern (UniProt P13688 subcellular location; HPA tissue IHC). Check morphology and controls before calling it CEACAM1: cytoplasmic vesicle membrane and secreted locations are also annotated, so a limited intracellular signal alone cannot establish an artefact (UniProt P13688 subcellular location). |
| Strong staining in cells listed as undetected, such as adipocytes or bronchial respiratory epithelium (HPA tissue IHC). | Treat this as discordant with those cell specific HPA observations, not proof that the specimen lacks CEACAM1. Cross reactivity or endogenous chromogenic activity are possible explanations (general IHC practice); HPA specifically cautions that its tissue staining may target protein from more than one gene (HPA tissue IHC). |
| A diffuse brown haze extends across tissue and surrounding areas. | A field wide deposit without cell border definition cannot be scored as the reported membranous pattern (HPA tissue IHC; general IHC practice). Examine a reagent control and the distribution of precipitate; background from detection chemistry or incomplete blocking is possible (general IHC practice). |
| No defined signal in colon or rectal enterocyte microvilli (HPA tissue IHC). | The expected High reference signal is absent (HPA tissue IHC). Review the run controls and slide morphology before interpreting the specimen as negative (general IHC practice). HPA's medium RNA to staining consistency and multi gene target caution limit how conclusively a single negative slide can be read (HPA tissue IHC). |
💡Expected CEACAM1 appearanceCall a result positive when enterocyte microvilli or cell borders show distinct membranous staining, potentially High in colon, rectum or appendix (HPA tissue IHC; UniProt P13688 subcellular location); diffuse haze or isolated nuclear deposits lack the expected pattern (general IHC practice).
| Membrane domain and epitope position (UniProt P13688 topology). | CEACAM1 spans residues 429–452, with an extracellular region at 35–428 and a cytoplasmic region at 453–526 (UniProt P13688 topology). Without a mapped antibody epitope, the record does not establish which region the stain detects. |
| Isoforms and processing (UniProt P13688). | UniProt lists 11 isoforms and a signal peptide at residues 1–34; the annotated mature chain begins at residue 35 (UniProt P13688 isoforms and processing). Antibody coverage across isoforms or processed forms cannot be inferred without its epitope. |
| Tissue and cell context (HPA tissue IHC). | HPA records High enterocyte microvillus staining in colon, rectum and appendix, Medium in duodenum and small intestine, and Low hepatocyte staining (HPA tissue IHC). Compare like cell types and compartments when judging relative intensity. |
| Antibody evidence (HPA antibodies; HPA tissue IHC). | The listed IHC antibodies have different validation labels: HPA011041 is Supported and CAB002146 is Enhanced (HPA antibodies). The overall tissue profile is Enhanced, with medium RNA to staining consistency and a multi gene target caution (HPA tissue IHC). These labels do not establish specificity for an individual slide. |
| Q: What should IF/ICC show? | A: Membrane localisation is the HPA subcellular summary, while UniProt also lists lateral and apical membranes and junctions (HPA subcellular; UniProt P13688 subcellular location). HPA supplies no ICC/IF image cell line or main location here, so an IF pattern cannot be independently checked against that record (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a reference intestinal section. | The reference expectation is High enterocyte microvillus staining in colon, rectum or appendix (HPA tissue IHC); the cause of its absence is undetermined. | Confirm the intended cells are present, inspect assay controls, and review retrieval and detection settings using the antibody's validated IHC instructions (general IHC practice). Do not infer CEACAM1 specific fixation sensitivity from these sources. |
| Only diffuse cytoplasmic colour appears. | It lacks the predominant membrane pattern (UniProt P13688 subcellular location; HPA tissue IHC), although UniProt also annotates a secretory vesicle membrane location (UniProt P13688). | Check whether any crisp apical or lateral border remains and compare with a reagent control before scoring the signal (general IHC practice). |
| Strong staining appears in an HPA undetected cell type. | Cross reactivity or endogenous detection activity may explain discordant colour (general IHC practice); HPA flags possible detection of protein from more than one gene (HPA tissue IHC). | Check a matched negative control and review cellular morphology and stain distribution; avoid assigning target identity from intensity alone (general IHC practice). |
| Brown deposits obscure epithelial borders. | Diffuse detection background or precipitate may hide the reported membranous pattern (general IHC practice; HPA tissue IHC). | Inspect a reagent control and background outside the target cells; review blocking, washing and chromogen development as general IHC workflow checks (general IHC practice). |
| A liver slide looks faint beside a colon slide. | HPA lists hepatocytes as Low and colon enterocyte microvilli as High; its profile specifically notes liver bile canaliculi (HPA tissue IHC). | Judge the liver's canalicular distribution with its own morphology and controls instead of expecting colon level intensity (HPA tissue IHC; general IHC practice). |
| Two antibodies yield different distributions. | HPA assigns Supported to HPA011041 and Enhanced to CAB002146, while warning that staining may target more than one gene (HPA antibodies; HPA tissue IHC). The reason for a specific discrepancy remains unknown. | Compare membrane localisation and cell types on matched sections, then report antibody identity and the disagreement alongside each result (HPA tissue IHC; general IHC practice). |
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: its enterocyte microvilli should stain strongly (HPA: Colon enterocytes – microvilli, High). Use adipose tissue adipocytes as the negative comparator (HPA: Adipocytes, Not detected); on the colon slide, compare the apical signal with nearby non-enterocyte cells as internal background, without assuming those cells are CEACAM1-negative (HPA: Colon enterocytes – microvilli, High; UniProt P13688: expression in granulocytes and lymphocytes).
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, an isotype-matched rabbit IgG control, and, if available, a CEACAM1 knockout specimen processed alongside the test section (caption: rabbit anti-CEACAM1 M00923-2; standard IHC controls). For chromogenic detection, quench endogenous peroxidase and assess background on the no-primary slide (caption: HRP and DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M00923-2 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption uses heat-mediated EDTA retrieval at pH 8.0, but does not establish whether retrieval is required (caption: M00923-2 tissue IHC). Relative ease of frozen-section IHC or IF/ICC is unreported; in colon, assess microvillar staining against tissue morphology and control-slide background (HPA: Colon enterocytes – microvilli, High).
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 epitope and cell specific controls (datasheet M00923-2; UniProt P13688 topology).
What retrieval should I try first when CEACAM1 staining is weak?
Start with heat mediated antigen retrieval in EDTA, pH 8.0, for paraffin sections (datasheet M00923-2). That is the retrieval used for the catalog antibody in human colorectal adenocarcinoma tissue; its caption gives no heating duration, so record and optimise that variable locally (datasheet M00923-2; standard IHC practice). Keep the primary incubation at the documented 1:50 overnight at 4°C while assessing retrieval, so changes remain interpretable (datasheet M00923-2; standard IHC practice). If staining remains weak, compare an alternative retrieval buffer on serial sections, using known positive tissue and a no primary control to assess signal and background (HPA: high in colon enterocyte microvilli; standard IHC practice).
Could fixation explain variable CEACAM1 staining across paraffin blocks?
The catalog image documents a paraffin section but does not state its fixative, so CEACAM1 sensitivity to fixation is unknown for this antibody (datasheet M00923-2 caption). Record each block’s fixative and fixation interval, then compare sections using the same EDTA, pH 8.0 retrieval and 1:50 primary dilution (datasheet M00923-2; standard IHC practice). Use a consistent positive control, such as colon epithelium, in each run and compare its membrane staining across blocks (HPA: high in colon enterocyte microvilli; standard IHC practice). If signal differs, optimise processing on matched material before attributing the difference to CEACAM1 expression; the supplied evidence establishes no target specific fixation effect (datasheet M00923-2 caption; standard IHC practice).
Where should convincing CEACAM1 chromogenic staining appear in tissue?
Prioritise staining along epithelial cell membranes, especially apical microvilli in intestinal epithelium and bile canaliculi in liver (HPA tissue IHC profile; UniProt P13688 subcellular location). CEACAM1 can also occupy lateral and basal membranes, cell junctions and secretory vesicle membranes, so document the compartment instead of reporting a whole cell positive result (UniProt P13688 subcellular location; standard IHC practice). Colon, rectum and appendix enterocyte microvilli are high staining reference sites, whereas hepatocyte staining is listed as low (HPA tissue IHC). If staining appears diffuse, review the counterstain and neighbouring structures at higher magnification, then compare a no primary control before scoring membrane positivity (standard IHC practice).
Can this stain distinguish CEACAM1 isoforms or cytoplasmic signalling states?
Do not assign an isoform from this chromogenic stain without a mapped antibody epitope and isoform specific validation; the record lists 11 CEACAM1 isoforms (UniProt P13688 isoforms; standard IHC practice). The full length topology places residues 35–428 extracellularly, 429–452 across the membrane and 453–526 cytoplasmically (UniProt P13688 topology). A cytoplasmic epitope could be affected by isoform structure or modification, including phosphorylation at residues 493 and 520, but this antibody’s epitope is not supplied (UniProt P13688 modified residues; datasheet M00923-2 caption). Report membrane staining and the antibody used; use a separately validated isoform or modification specific assay for stronger assignments (standard IHC practice).
How can I follow up an IHC pattern with multiplex CEACAM1 immunofluorescence?
Treat IF as a separate assay requiring its own antibody validation; the supplied catalog example establishes paraffin section chromogenic IHC conditions only (datasheet M00923-2 caption; standard IF practice). Pair CEACAM1 with an independently validated marker for the expected cell type, such as an enterocyte marker when investigating intestinal microvilli, and assess channels separately before interpreting overlap (HPA: high in colon enterocyte microvilli; standard IF practice). Choose fluorophores after checking tissue autofluorescence and include single stain and no primary controls for bleed through and background (standard IF practice). If the validated epitope is extracellular, minimise permeabilisation; if it is cytoplasmic, optimise permeabilisation while preserving membrane structure, since this antibody’s epitope location is unspecified (UniProt P13688 topology; standard IF practice).
How should I troubleshoot diffuse DAB signal around CEACAM1 positive glands?
First compare a no primary control with the stained section and check whether DAB appears outside the expected epithelial membrane pattern (HPA tissue IHC profile; standard IHC practice). Include a peroxidase blocking step in the chromogenic workflow and examine endogenous pigment or precipitate before calling diffuse colour CEACAM1; these are general IHC checks (standard IHC practice). The catalog example used 10% goat serum block, 1:50 primary overnight at 4°C, and a peroxidase secondary for 30 minutes at 37°C (datasheet M00923-2). If background persists, optimise blocking, antibody concentration and washing one variable at a time while retaining a positive tissue control (standard IHC practice).
How should I score CEACAM1 across sections with different epithelial content? ⚠ ANSWER MARKED FOR VERIFICATION
Define the epithelial region before scoring and exclude necrotic or damaged areas by a prespecified rule (standard IHC practice). For an intensity aware summary, report an H-score from 0–300 using the percentages of cells at intensities 0–3; also report the percentage of membrane positive epithelial cells (standard IHC practice). Normalise counts to the number of evaluable epithelial cells, or report positive cell density per mm² of annotated epithelial area, rather than per whole section area (standard IHC practice). Record apical and lateral membrane staining separately when relevant, and keep exposure, counterstain and scoring thresholds consistent across batches (UniProt P13688 subcellular location; standard IHC practice).
Which findings support genuine CEACAM1 signal rather than staining artefact?
A convincing positive follows a plausible membrane compartment, such as intestinal enterocyte microvilli or hepatocyte canaliculi, and exceeds the matched no primary control (HPA tissue IHC profile; standard IHC practice). Question isolated nuclear colour or diffuse staining in a cell type without supporting expression evidence; CEACAM1 is described chiefly at membranes and junctions (UniProt P13688 subcellular location; standard IHC practice). Discount section edge deposits, necrotic areas and colour reproduced by the no primary control or endogenous peroxidase when interpreting DAB (standard IHC practice). HPA reports Enhanced reliability but cautions that its staining may detect protein from more than one gene, so confirm a consequential assignment independently (HPA tissue IHC reliability description).
Boster reagents
Best CEACAM1 / Cell adhesion molecule CEACAM1 IHC Antibodies
The human-reactive catalog antibody has real IHC data from paraffin-embedded colorectal adenocarcinoma and liver cancer sections (catalog: Human reactivity; M00923-2 IHC captions). IF is listed without an IF image (catalog: applications and image records).
Real IHC data
Anti-CEACAM1 Rabbit Monoclonal Antibody
Cat # M00923-2
M00923-2 has IHC images from human paraffin-embedded colorectal adenocarcinoma and liver cancer sections (M00923-2 IHC captions). IF/ICC is listed as an application, but no IF image is provided (catalog: M00923-2 applications and image records).
Which to pick: Choose M00923-2 for human paraffin-section IHC: it is a rabbit monoclonal, and its captions document EDTA pH 8.0 retrieval, 1:50 primary dilution, and DAB detection; the fixative is unreported (catalog: M00923-2 host and clone; M00923-2 IHC captions). For IF/ICC, M00923-2 is the listed option with a catalog dilution of 1:50, but no IF image is provided (catalog: M00923-2 applications, dilution and image records). No cross-species option is documented; M00923-2 lists Human reactivity only (catalog: M00923-2 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 Membranous staining in GI glands and bile canaliculi (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 fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may reflect protein from more than one gene (HPA tissue IHC); Isoform / epitope: 11 isoforms; check extracellular versus cytoplasmic epitopes (UniProt); Regulation: Expression regulation is not 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.