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- Table of Contents
HOXA3 / Homeobox protein Hox-A3 · Western blot design guide
Design a Western Blot for HOXA3
Source-linked HOXA3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HOXA3 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
HOXA3 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 | ~46.4 kDa | |
| Observed band | 46 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 HOXA3 Western Blot Protocol Options
The PA1602 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 | 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 | PA1602; 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 HOXA3 Western Blot Band Size?
HOXA3 is predicted at 46.4 kDa and observed at 46 kDa; the small difference has no established cause in the supplied evidence.
| Band at 46 kDa in whole-cell lysate | matches the reported HOXA3 antibody QC band |
| Single band near 46 kDa | consistent with the 46.4 kDa predicted mass |
| 46 kDa band enriched in a nuclear fraction | consistent with HOXA3 nuclear localization |
| Weak 46 kDa band in whole-cell lysate | nuclear enrichment may improve detection |
💡Expected HOXA3 appearanceHOXA3 has a predicted mass of 46.4 kDa and an empirical 46 kDa band in whole-cell lysate; confirm band identity with antibody specificity and nuclear fraction controls.
| UniProt predicted mass | places full-length HOXA3 near 46.4 kDa |
| Predicted 46,369 Da mass | corresponds to approximately 46.4 kDa |
| 443-residue HOXA3 sequence | underlies the reported full-length mass prediction |
| Full-length HOXA3 mass prediction | closely matches the observed 46 kDa band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear HOXA3 may be poorly recovered | check a nuclear fraction and lysate preparation |
| Band higher than expected | band identity is uncertain | compare with the 46 kDa QC band and validate antibody specificity |
| Band lower than expected | band identity is uncertain | check antibody specificity and sample integrity |
| Multiple bands | additional bands have no assigned HOXA3 feature here | identify the 46 kDa band with specificity controls |
| Weak or no signal | nuclear HOXA3 may be diluted in whole-cell lysate | test a nuclear fraction and a positive control |
| Fragments below expected size | sample degradation is possible | check sample handling and compare fresh lysate |
Experiment evidence
Western blot images showing HOXA3 band
PA1602 · HOXA3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HOXA3 Western blot
🧪For positive controls for HOXA3 in Western blot, you can use no HPA-supported tissue or cell line from the supplied evidence.
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: HOXA3 is nuclear, but the supplied HPA data identify no tissue controls.
HPA tissue expression evidence for HOXA3
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 HOXA3 Western Blot Tips
Deeper troubleshooting and optimisation questions for HOXA3, answered from its protein features.
How should HOXA3 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 HOXA3 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no basis for assigning an additional band to an HOXA3 isoform.
Can an annotated modification explain a shifted HOXA3 band?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not identify a modification that explains a shift; they also cannot rule out every possible modification.
Does this guide establish induction of HOXA3?
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 HOXA3?
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 PA1602 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 the HOXA3 band be quantified?
Quantitation · Quantify the band near 46 kDa consistently across comparable samples. Because HOXA3 is nuclear, use the same sample preparation and a consistent loading reference for each comparison.
Does the observed HOXA3 band match its predicted mass?
Interpretation · The reported band is 46 kDa, close to the predicted 46.4 kDa for the 443-residue protein. This agreement does not establish band identity by itself.
HOXA3 is annotated in the nucleus. Include a nuclear sample when assessing its 46 kDa band, and compare like sample preparations across lanes.
Check the band near the reported 46 kDa position first. The supplied record lists one isoform and no annotated modified residues, so it does not support assigning other bands to specific isoforms or modifications.
HOXA3 is annotated in the nucleus. Include a nuclear sample when assessing its 46 kDa band, and compare like sample preparations across lanes.
Check the band near the reported 46 kDa position first. The supplied record lists one isoform and no annotated modified residues, so it does not support assigning other bands to specific isoforms or modifications.
Boster reagents
HOXA3 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-A3 HOXA3 Antibody Picoband®
Cat # PA1602
PA1602 is listed for human, mouse, and rat HOXA3. Its Western blot image shows a 46 kDa band in SW620 whole cell lysate, matching the predicted size. The supplied image documents this sample and condition only.
Which to pick: PA1602 is the only listed antibody. It has a Western blot image using SW620 whole cell lysate at 40 µg and antibody at 0.5 µg/mL; the supplied evidence does not show mouse or rat blots.
Source: BosterBio HOXA3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43365.
- Human Protein Atlas. HOXA3 tissue expression.
- PMC7585938 — target-verified WB comparison
- PMC8034473 — target-verified WB comparison
- PMC12894161 — target-verified WB comparison