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- Table of Contents
DLX1 / Homeobox protein DLX-1 · Western blot design guide
Design a Western Blot for DLX1
Real validated DLX1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DLX1 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
DLX1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~27.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Testis (IHC candidate; verify WB) +1 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
Real Curated DLX1 Western Blot Protocols
The A07125 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A07125; 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 DLX1 Western Blot Band Size?
DLX1 is predicted at 27.3 kDa; two isoforms could affect migration, but no empirical band size or distinct isoform bands are established.
| Band near 27.3 kDa | Consistent with the predicted DLX1 mass; confirm identity with controls |
| One band despite two annotated isoforms | Isoforms 1 and 2 are not established to migrate separately |
| Two bands at different positions | Could reflect isoforms 1 and 2; their migration has not been established |
| Weak whole-cell lysate band | Nuclear DLX1 may be underrepresented in the tested preparation |
💡Expected DLX1 appearanceDLX1 has a predicted mass of 27.3 kDa, but no empirical band size is supplied; use band-identity controls because the migration of isoforms 1 and 2 is unknown.
| UniProt predicted mass | Places the reference full-length protein near 27.3 kDa |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | May produce different apparent sizes, but no size difference is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear DLX1 may be underrepresented in the preparation | Check a nuclear-enriched fraction and a positive-control sample |
| Band higher than expected | The identity and migration of the annotated isoforms are unknown | Compare with an independent antibody or DLX1-depleted control |
| Band lower than expected | The identity and migration of the annotated isoforms are unknown | Check band identity with an independent antibody or DLX1-depleted control |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are unverified | Compare isoform expression and use a DLX1-depleted control |
| Weak or no signal | Nuclear DLX1 may be poorly represented in the tested sample | Test a nuclear-enriched fraction and a positive-control sample |
Experiment evidence
Western blot images showing DLX1 band
A07125 · DLX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DLX1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DLX1 in Western blot, you can use testis lysate, which has high HPA expression.
Positive control: Testis (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: DLX1 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for DLX1
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 |
|---|---|---|---|---|
| Testis | elongated or late spermatids | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DLX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DLX1, answered from its protein features.
How should DLX1 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 DLX1 isoforms produce different bands?
Isoforms · UniProt lists two isoforms. In UniProt canonical coordinates, isoform 2 replaces residues 106–255 with QDLVPKQAIQVQEADEAGWGGSGG, so its sequence is shorter. Distinct bands are possible, but no observed band sizes are supplied. Confirm which isoforms your antibody recognizes.
Choose an epitope within residues 1–105 of the canonical UniProt sequence, which precede the isoform 2 replacement. An antibody against canonical residues 106–255 may miss isoform 2. Check the antibody’s stated epitope and isoform reactivity.
Decide whether the measurement is for total DLX1 or a specific isoform. UniProt lists two isoforms with different sequences, so confirm antibody coverage before combining band signals. Report which bands were measured and normalize consistently across samples.
Choose an epitope within residues 1–105 of the canonical UniProt sequence, which precede the isoform 2 replacement. An antibody against canonical residues 106–255 may miss isoform 2. Check the antibody’s stated epitope and isoform reactivity.
Decide whether the measurement is for total DLX1 or a specific isoform. UniProt lists two isoforms with different sequences, so confirm antibody coverage before combining band signals. Report which bands were measured and normalize consistently across samples.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of DLX1?
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.
What transfer method to use for DLX1 Western blot?
Transfer · The supplied features specify a 27.3 kDa predicted mass but no transfer method. Check that protein near this size transfers and remains on the membrane under your chosen conditions; the features do not justify a specific membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07125 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 DLX1 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.
Why might DLX1 migrate away from its predicted mass?
Interpretation · UniProt gives a predicted mass of 27.3 kDa, but no observed Western blot band is supplied. The listed features do not establish a visible shift or explain any difference between apparent and predicted mass. Compare your band with a molecular weight marker and validate its identity.
UniProt places DLX1 in the nucleus. A nuclear fraction is therefore relevant when checking for its signal; compare it with your input if fractionation is used. Nuclear localization alone does not establish the expected band intensity.
Check whether the antibody recognizes both listed isoforms, since isoform 2 replaces canonical residues 106–255 in UniProt numbering. UniProt lists no modified residues or glycosylation sites, and no empirical band position is provided. Band position alone cannot identify an isoform or establish a modification.
UniProt places DLX1 in the nucleus. A nuclear fraction is therefore relevant when checking for its signal; compare it with your input if fractionation is used. Nuclear localization alone does not establish the expected band intensity.
Check whether the antibody recognizes both listed isoforms, since isoform 2 replaces canonical residues 106–255 in UniProt numbering. UniProt lists no modified residues or glycosylation sites, and no empirical band position is provided. Band position alone cannot identify an isoform or establish a modification.
Boster reagents
DLX1 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 DLX-1 DLX1 Antibody
Cat # A07125
A07125 is an anti-DLX1 antibody listed for Western blotting with Human, Mouse, and Rat reactivity. Its WB image shows DU145 cell lysate tested at a 1:1000 primary antibody dilution overnight at 4°C; the supplied evidence does not show other specimens.
Which to pick: A07125 is the only listed DLX1 antibody. It has a WB image from DU145 cell lysate; check whether that sample and the listed reactivity fit your experiment.
Source: BosterBio DLX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P56177.
- Human Protein Atlas. DLX1 tissue expression.
- PMC6714137 — target-verified WB comparison
- PMC7980590 — target-verified WB comparison
- PMC5348575 — target-verified WB comparison