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- Table of Contents
EMX2 / Homeobox protein EMX2 · Western blot design guide
Design a Western Blot for EMX2
Source-linked EMX2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-EMX2 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
EMX2 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 | ~28.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebellum (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 EMX2 Western Blot Protocol Options
The A04322-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human lung tissue lysate (catalog A04322-1) |
| 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 | A04322-1 · (A) 1 and (B) 2 μg/mL (catalog A04322-1) |
| 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 EMX2 Western Blot Band Size?
EMX2 is predicted at 28.3 kDa; its two isoforms could affect migration, but no distinct isoform bands or empirical band size are established.
| Single band near 28.3 kDa | Consistent with the predicted EMX2 mass; confirm identity with antibody controls |
| Two discrete bands at different positions | Could reflect isoforms 1 and 2 if both are expressed and resolve separately |
| Single band away from 28.3 kDa | Could represent an EMX2 isoform, but its identity requires confirmation |
| Faint band in whole-cell lysate | Nuclear localization may make EMX2 easier to detect in an enriched fraction |
💡Expected EMX2 appearanceEMX2 has a predicted mass of 28.3 kDa, but no empirical band size or isoform migration is supplied; use antibody specificity controls to identify any detected band.
| UniProt predicted mass | Places the reference size at 28.3 kDa |
| Alternative splicing | Could produce products with different apparent sizes; separation is unestablished |
| Isoform 1 | Its size relative to isoform 2 is not supplied |
| Isoform 2 | Its size relative to isoform 1 is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear EMX2 may be difficult to detect in an unfractionated sample | Check a nuclear fraction and verify antibody performance |
| Band higher than expected | An isoform is possible, but its migration is not established | Compare with an EMX2-positive control and verify antibody specificity |
| Band lower than expected | An isoform is possible, but its migration is not established | Use an EMX2-positive control and check for sample degradation |
| Multiple bands | Isoforms 1 and 2 may contribute, though distinct bands are not established | Check antibody specificity and compare samples with different EMX2 expression |
| Weak or no signal | Nuclear localization may limit signal in the tested fraction | Enrich nuclei and check lysate loading and antibody performance |
| Fragments below expected size | No cleavage feature is listed to explain fragments | Check sample integrity and confirm band identity with specificity controls |
Experiment evidence
Western blot images showing EMX2 band
A04322-1 · EMX2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EMX2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for EMX2 in Western blot, you can use cerebellum lysate, based on HPA detection in cerebellum.
Positive control: Cerebellum (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 EMX2.
⚠️Feasibility: Because EMX2 is nuclear, a nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for EMX2
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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - cytoplasm/membrane | Medium | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | non-ciliated cells | Medium | Protein (IHC) | HPA → |
| Kidney | collecting ducts | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced EMX2 Western Blot Tips
Deeper troubleshooting and optimisation questions for EMX2, answered from its protein features.
How should EMX2 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 EMX2 isoforms produce separate bands?
Isoforms · UniProt lists two isoforms. In isoform 2, canonical residues 137–252 are replaced by a shorter sequence, so the isoforms may differ in mass. Their annotation alone does not establish that two bands will be visible.
Choose an epitope within canonical residues 1–136, which precede the sequence replacement in isoform 2. An antibody against canonical residues 137–252 may miss isoform 2.
Decide whether the measurement is total EMX2 or isoform-specific. For total EMX2, use an antibody recognizing the shared residues 1–136 and account for any separately resolved bands. For isoform-specific measurement, verify that the antibody distinguishes the altered region.
Choose an epitope within canonical residues 1–136, which precede the sequence replacement in isoform 2. An antibody against canonical residues 137–252 may miss isoform 2.
Decide whether the measurement is total EMX2 or isoform-specific. For total EMX2, use an antibody recognizing the shared residues 1–136 and account for any separately resolved bands. For isoform-specific measurement, verify that the antibody distinguishes the altered region.
Can annotated modifications explain an EMX2 band shift?
PTM · The supplied UniProt record lists no modified residues or glycosylation sites. It therefore provides no annotated modification that explains a shifted band. A band shift cannot be assigned a cause from these features alone.
Does this guide establish induction of EMX2?
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 EMX2?
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 A04322-1 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 EMX2 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
What size should EMX2 show on Western blot?
Interpretation · The canonical EMX2 sequence is 252 residues with a predicted mass of 28.3 kDa. No observed band size is supplied, so use 28.3 kDa as a reference, not an expected exact position on the blot.
UniProt annotates EMX2 in the nucleus and in axons, based on evidence transferred from a related protein. A nuclear fraction is a reasonable starting point; consider an axon-containing sample when that location is relevant to the experiment.
Check whether a band could reflect isoform 2, whose canonical residues 137–252 are replaced. Compare detection with antibodies against shared and altered regions if available. The record supplies no observed band size or annotated modification that would identify an unexpected band by position alone.
UniProt annotates EMX2 in the nucleus and in axons, based on evidence transferred from a related protein. A nuclear fraction is a reasonable starting point; consider an axon-containing sample when that location is relevant to the experiment.
Check whether a band could reflect isoform 2, whose canonical residues 137–252 are replaced. Compare detection with antibodies against shared and altered regions if available. The record supplies no observed band size or annotated modification that would identify an unexpected band by position alone.
Boster reagents
EMX2 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-Homeobox protein EMX2 EMX2 Antibody
Cat # A04322-1
The listed anti-EMX2 antibody has a Western blot image using human lung tissue lysate at 1 and 2 μg/mL. The supplied evidence covers this tested context only.
Which to pick: A04322-1 is the only listed option. It reports human reactivity and includes a Western blot image from human lung tissue lysate; consider your sample and assay conditions when choosing it.
Source: BosterBio EMX2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q04743.
- Human Protein Atlas. EMX2 tissue expression.
- PMC11614513 — target-verified WB comparison
- PMC8106024 — target-verified WB comparison
- PMC5568393 — target-verified WB comparison