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- Table of Contents
HOXD13 / Homeobox protein Hox-D13 · Western blot design guide
Design a Western Blot for HOXD13
Source-linked HOXD13 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HOXD13 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
HOXD13 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 | ~36.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked HOXD13 Western Blot Protocol Options
The A03000 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | The brain of Mouse(30ug), The brain of Rat(30ug) (catalog A03000) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03000; 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 HOXD13 Western Blot Band Size?
HOXD13 is predicted at 36.1 kDa; no empirical band size or feature-driven migration difference is established.
| Single band near 36.1 kDa | Consistent with the predicted HOXD13 mass; identity requires confirmation |
| Band enriched in a nuclear fraction | Consistent with HOXD13's nuclear location |
| Weak band in a cytoplasmic fraction | Consistent with HOXD13's nuclear location |
| Single band in a nuclear extract | Consistent with the one annotated HOXD13 isoform |
💡Expected HOXD13 appearanceHOXD13 is predicted at 36.1 kDa, but no empirical band size is supplied; verify a candidate band with an independent antibody or HOXD13 depletion.
| UniProt predicted mass of 36.1 kDa | Provides the reference size for full-length HOXD13 |
| Predicted full-length molecular weight | Suggests a band near 36.1 kDa if migration reflects calculated mass |
| Calculated molecular weight of 36,101 Da | Corresponds to a predicted size of about 36.1 kDa |
| Predicted HOXD13 reference size | Does not establish an empirical migration position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HOXD13 may be poorly recovered | Check nuclear extraction with a nuclear protein control |
| Band higher than expected | No annotated feature establishes a higher migrating band | Check identity with HOXD13 depletion or an independent antibody |
| Band lower than expected | The supplied features do not establish a smaller HOXD13 product | Compare antibodies recognizing different regions |
| Multiple bands | The single annotated isoform does not explain distinct bands | Determine which bands respond to HOXD13 depletion |
| Weak or no signal | HOXD13 may be scarce in the tested sample | Check sample expression and nuclear recovery |
Experiment evidence
Western blot images showing HOXD13 band
A03000 · HOXD13
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HOXD13 Western blot
🧪For positive controls for HOXD13 in Western blot, you can use no HPA-supported tissue or cell sample because none is reported.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue expression data, so a positive sample cannot be selected and a knockdown or KO is needed for a negative control.
HPA tissue expression evidence for HOXD13
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced HOXD13 Western Blot Tips
Deeper troubleshooting and optimisation questions for HOXD13, answered from its protein features.
Where should the HOXD13 band appear?
Band shift · The predicted mass is 36.1 kDa for the 343-residue protein. No observed band position is supplied, so use 36.1 kDa as a reference rather than an expected apparent mass.
Could an isoform explain a second band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for a second band.
Do annotated modifications predict a band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not predict a modification-related shift, but they cannot establish why an observed band differs from 36.1 kDa.
Does this guide establish induction of HOXD13?
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 HOXD13?
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 A03000 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 HOXD13 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.
Which sample fraction is most relevant for detection?
Interpretation · HOXD13 is annotated as nuclear. Check a nuclear fraction when evaluating a weak signal, and compare fractions using an appropriate loading control.
Because HOXD13 is annotated as nuclear, compare consistently prepared fractions and normalize the signal to a suitable control for the fraction measured. The supplied record gives no induction condition or expected change in abundance.
The record lists disease variant and triplet repeat expansion keywords, but gives no variant sequence, repeat length, or observed band size. Those keywords alone cannot identify an unexpected band or establish a mass shift; confirm band identity independently.
Because HOXD13 is annotated as nuclear, compare consistently prepared fractions and normalize the signal to a suitable control for the fraction measured. The supplied record gives no induction condition or expected change in abundance.
The record lists disease variant and triplet repeat expansion keywords, but gives no variant sequence, repeat length, or observed band size. Those keywords alone cannot identify an unexpected band or establish a mass shift; confirm band identity independently.
Boster reagents
HOXD13 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 Hox-D13 HOXD13 Antibody
Cat # A03000
A03000 is an anti-HOXD13 polyclonal antibody listed for human, mouse, and rat. Its WB image shows mouse and rat brain tissue lysates (30 µg per lane) at 1:500 dilution. The supplied evidence does not show a human WB sample.
Which to pick: A03000 is the only listed option. Its WB image supports choosing it for mouse or rat brain lysates under the reported conditions; human reactivity is listed, but no human WB example is supplied.
Source: BosterBio HOXD13 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P35453.
- Human Protein Atlas. HOXD13 tissue expression.
- PMC12218409 — target-verified WB comparison
- PMC12232685 — target-verified WB comparison
- PMC8366098 — target-verified WB comparison