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CEACAM6 / Cell adhesion molecule CEACAM6 · Western blot design guide
Design a Western Blot for CEACAM6
Source-linked CEACAM6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CEACAM6 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
CEACAM6 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~37.2 kDa | |
| Observed band | ~60 kDa | |
| Gel | 10% (catalog M03197-1) | |
| Positive control | Appendix (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CEACAM6 Western Blot Protocol Options
The M03197-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, mouse brain (catalog M03197-1) |
| Gel % | 10% (catalog M03197-1) |
| Load | 30 ug; reducing conditions (catalog M03197-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03197-1) |
| Membrane | nitrocellulose membrane (catalog M03197-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M03197-1) |
| Primary antibody | M03197-1 · 1:1000 (catalog M03197-1) |
| Primary incubation | overnight at 4°C (catalog M03197-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M03197-1) |
| Secondary incubation | 1.5 hour at RT (catalog M03197-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M03197-1) |
| Detection | ECL (catalog M03197-1) |
Section 2
What Is the Expected CEACAM6 Western Blot Band Size?
CEACAM6 has a predicted precursor mass of 37.2 kDa and an empirical band near 60 kDa; the cause of their difference is not established.
| Band near 60 kDa | Empirical CEACAM6 band reported in reducing Western blots; confirm identity with controls |
| Band near 37.2 kDa | Possible species near the predicted precursor mass; identity needs confirmation |
| Band below the precursor position | Could reflect signal-peptide or propeptide cleavage |
| Broad band or smear | Could reflect variable N-linked glycosylation |
| Higher band under nonreducing conditions | Could reflect a CEACAM6 homodimer if it survives electrophoresis |
💡Expected CEACAM6 appearanceUniProt predicts a 37.2 kDa precursor, while antibody QC reports a band near 60 kDa in reducing blots; the cause of the difference is unproven, so confirm identity with appropriate controls.
| Predicted precursor mass | 37.2 kDa is the sequence-based reference, not the measured migration |
| N-linked glycosylation at Asn104, Asn111, Asn115, Asn152, Asn173, Asn197, Asn224, and Asn256 | May increase or vary apparent mass; the size of any shift is not established |
| Homodimerization through the Ig-like V-type domain | May produce a higher band if the complex remains intact during electrophoresis |
| Signal-peptide cleavage at residues 1–34 | Removes sequence from the precursor, potentially lowering mature-protein mass |
| Propeptide cleavage at residues 321–344 | Removes sequence from the precursor, potentially lowering mature-protein mass |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CEACAM6 is GPI-anchored at the cell surface and may be poorly represented in the sampled lysate | Check a CEACAM6-positive control and enrich the membrane fraction |
| Band higher than expected | N-linked glycosylation or a homodimer that survives sample preparation may contribute; the 60 kDa difference is not established | Compare reducing conditions and assess glycosylation with suitable controls |
| Band lower than expected | Signal-peptide or propeptide processing may remove precursor sequence | Compare with a positive control and confirm band identity by CEACAM6 depletion |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Assess glycosylation and compare with a CEACAM6-positive control |
| Multiple bands | Processing or variable glycosylation may yield different species | Use CEACAM6 depletion and appropriate processing or glycosylation controls |
| Weak or no signal | The sampled fraction may contain little cell-surface CEACAM6 | Verify expression with a positive control and test a membrane-enriched fraction |
Experiment evidence
Western blot images showing CEACAM6 band
M03197-1 · CEACAM6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CEACAM6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CEACAM6 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: CEACAM6 is GPI-anchored at the cell surface, so use a lysate that retains membrane proteins.
HPA tissue expression evidence for CEACAM6
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Colon | enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | Medium | Protein (IHC) | HPA → |
| Rectum | enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CEACAM6 Western Blot Tips
Deeper troubleshooting and optimisation questions for CEACAM6, answered from its protein features.
How should CEACAM6 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could alternative CEACAM6 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis to assign multiple bands to annotated isoforms. Compare bands across samples and assess whether processing or glycosylation could be relevant before assigning their identity.
Which CEACAM6 glycosylation sites should I consider?
PTM · UniProt annotates N-linked sites at residues 104, 111, 115, 152, 173, 197, 224, 256, 274, 288, 292, and 309. These are UniProt sequence coordinates; antibody or paper numbering may differ. The annotations do not establish occupancy in your sample or predict a specific band shift.
Does this guide establish induction of CEACAM6?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for CEACAM6?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03197-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should I prepare samples for CEACAM6 quantitation?
Quantitation · CEACAM6 is annotated at the cell surface, including the apical membrane, with a GPI anchor. Use the same sample preparation and loading basis across samples so differences in membrane recovery do not distort the comparison. Quantify the same identified band consistently.
Why might CEACAM6 appear near 60 kDa instead of 37.2 kDa?
Interpretation · The 37.2 kDa prediction is based on the supplied sequence, while ~60 kDa is the reported apparent band. CEACAM6 has 12 annotated N-linked glycosylation sites and undergoes terminal processing. These features may affect migration, but they do not establish the cause or size of the difference.
UniProt marks a signal peptide at residues 1–34 and a propeptide at 321–344, and locates CEACAM6 at the cell surface with a GPI anchor. These UniProt coordinates describe processing regions; they do not by themselves predict the apparent mass of the mature band.
CEACAM6 can homodimerize and heterodimerize with CEACAM8 through its Ig-like V-type domain; UniProt also lists two disulfide bonds. Those features make association relevant to investigate, but they do not show that a particular Western-blot band is a dimer.
Check whether its pattern is consistent across samples and with the reported ~60 kDa band. Consider the annotated glycosylation sites, terminal processing regions, and possible association with CEACAM6 or CEACAM8. None of these features alone identifies an unexpected band or proves why it migrates at a given mass.
UniProt marks a signal peptide at residues 1–34 and a propeptide at 321–344, and locates CEACAM6 at the cell surface with a GPI anchor. These UniProt coordinates describe processing regions; they do not by themselves predict the apparent mass of the mature band.
CEACAM6 can homodimerize and heterodimerize with CEACAM8 through its Ig-like V-type domain; UniProt also lists two disulfide bonds. Those features make association relevant to investigate, but they do not show that a particular Western-blot band is a dimer.
Check whether its pattern is consistent across samples and with the reported ~60 kDa band. Consider the annotated glycosylation sites, terminal processing regions, and possible association with CEACAM6 or CEACAM8. None of these features alone identifies an unexpected band or proves why it migrates at a given mass.
Boster reagents
CEACAM6 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-CEACAM6 Rabbit Monoclonal Antibody
Cat # M03197-1
The catalog reports one anti-CEACAM6 antibody, M03197-1, with reported human, mouse, and rat reactivity. Its WB image uses rat and mouse brain lysates; the reported band is approximately 60 kDa, versus an expected 37 kDa.
Which to pick: M03197-1 is the only listed option and has a WB image from rat and mouse brain lysates. Human reactivity is listed, but this image does not show a human sample. Consider the reported band-size difference when interpreting results.
Source: BosterBio CEACAM6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P40199.
- Human Protein Atlas. CEACAM6 tissue expression.
- PMC8226832 — target-verified WB comparison
- PMC2778047 — target-verified WB comparison
- PMC3515475 — target-verified WB comparison