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- Table of Contents
CDX1 / Homeobox protein CDX-1 · Western blot design guide
Design a Western Blot for CDX1
Source-linked CDX1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CDX1 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
CDX1 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 | ~28.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Colon (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CDX1 Western Blot Protocol Options
The M04522-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Caco-2 cell lysate (catalog M04522-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M04522-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected CDX1 Western Blot Band Size?
CDX1 is predicted at 28.1 kDa; its two isoforms could affect migration, but no empirical band size or resolved isoform pattern is supplied.
| Band near 28.1 kDa | Consistent with the predicted CDX1 mass; confirm identity with controls |
| Two discrete bands | Could reflect isoforms 1 and 2 if their migration differs; confirm each band |
| One band despite two isoforms | The isoforms may comigrate or one may predominate |
| Stronger band in a nuclear fraction | Consistent with CDX1 nuclear localization |
💡Expected CDX1 appearanceCDX1 has a predicted mass of 28.1 kDa, but no empirical band size is supplied; isoforms 1 and 2 may affect migration, so confirm band identity with appropriate controls.
| UniProt predicted mass | Places the reference band near 28.1 kDa |
| Splice isoform 1 | May differ in apparent size from isoform 2; its mass is unspecified |
| Splice isoform 2 | May differ in apparent size from isoform 1; its mass is unspecified |
| Alternative splicing | May produce different band positions if the isoforms resolve |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear CDX1 may be dilute in whole-cell lysate | Check a nuclear fraction and a CDX1-positive control |
| Band higher than expected | Isoform migration or an unrelated band; the cause is unestablished | Compare isoform expression and verify identity with CDX1 depletion |
| Band lower than expected | Isoform migration or degradation; the cause is unestablished | Use fresh lysate with protease inhibitors and verify identity with CDX1 depletion |
| Multiple bands | Isoforms 1 and 2 may resolve, or some bands may be unrelated | Check which bands respond to CDX1 depletion |
| Weak or no signal | Low CDX1 abundance in the sampled lysate | Check a nuclear fraction and a CDX1-positive control |
| Fragments below expected size | Possible sample degradation, without a documented CDX1 cleavage feature | Prepare fresh lysate with protease inhibitors and compare with a CDX1-positive control |
Experiment evidence
Western blot images showing CDX1 band
M04522-1 · CDX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CDX1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CDX1 in Western blot, you can use colon tissue lysate.
Positive control: Colon (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Colon is a strong positive candidate; nuclear enrichment may help detect CDX1.
HPA tissue expression evidence for CDX1
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 |
|---|---|---|---|---|
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | endocrine cells | High | Protein (IHC) | HPA → |
| Small intestine | goblet cells | High | Protein (IHC) | HPA → |
| Appendix | endocrine cells | Medium | Protein (IHC) | HPA → |
| Duodenum | goblet cells | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CDX1, answered from its protein features.
How should CDX1 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 CDX1 isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 lacks residues 1–135 in UniProt numbering, so it has a different sequence from isoform 1. The supplied features do not establish where either isoform migrates on a blot.
An epitope within UniProt residues 1–135 is absent from isoform 2. An epitope in the retained region could permit detection of both isoforms. Check the antibody’s mapped epitope before interpreting a missing or additional band.
An epitope within UniProt residues 1–135 is absent from isoform 2. An epitope in the retained region could permit detection of both isoforms. Check the antibody’s mapped epitope before interpreting a missing or additional band.
Do annotated modifications explain a CDX1 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not support assigning a band shift to a specific modification, and feature annotations alone would not establish a visible shift.
Does this guide establish induction of CDX1?
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 CDX1?
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 M04522-1 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 CDX1 bands be quantified across samples?
Quantitation · Quantify the same candidate band under consistent sample preparation and normalization conditions. If multiple bands appear, assess them separately until their identities are established, since the two listed isoforms differ in sequence.
What mass is predicted for canonical CDX1?
Interpretation · The supplied predicted mass is 28.1 kDa. No observed band position is available, so use 28.1 kDa as a reference rather than an expected exact migration position.
CDX1 is listed as nuclear. Comparing a nuclear fraction with whole-cell material can help assess whether a candidate band follows that localization, although localization alone cannot identify the band.
Compare it with the 28.1 kDa prediction, the isoform 2 deletion of UniProt residues 1–135, antibody epitope location, and nuclear localization. The supplied features alone cannot identify an unexpected band or establish why its apparent mass differs.
CDX1 is listed as nuclear. Comparing a nuclear fraction with whole-cell material can help assess whether a candidate band follows that localization, although localization alone cannot identify the band.
Compare it with the 28.1 kDa prediction, the isoform 2 deletion of UniProt residues 1–135, antibody epitope location, and nuclear localization. The supplied features alone cannot identify an unexpected band or establish why its apparent mass differs.
Boster reagents
CDX1 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-Cdx1 Rabbit Monoclonal Antibody
Cat # M04522-1
The catalog reports one anti-CDX1 rabbit monoclonal antibody, M04522-1, with reported human reactivity. Its Western blot image shows Cdx1 expression in Caco-2 cell lysate; the supplied evidence does not establish performance in other samples.
Which to pick: M04522-1 is the only listed option. It has a Western blot image using Caco-2 cell lysate and reported human reactivity; assess suitability for your sample and conditions accordingly.
Source: BosterBio CDX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P47902.
- Human Protein Atlas. CDX1 tissue expression.
- PMC6356049 — target-verified WB comparison
- PMC3547218 — target-verified WB comparison
- PMC11929911 — target-verified WB comparison