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- Table of Contents
HOXA2 / Homeobox protein Hox-A2 · Western blot design guide
Design a Western Blot for HOXA2
Source-linked HOXA2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HOXA2 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
HOXA2 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 | ~41 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 HOXA2 Western Blot Protocol Options
The A05671 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A05671) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05671) |
| Primary antibody | A05671 · 1:1000 (catalog A05671) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05671) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05671) |
Section 2
What Is the Expected HOXA2 Western Blot Band Size?
HOXA2 is predicted at 41 kDa; no empirical band is supplied, and its listed features do not demonstrate a migration shift.
| Single sharp band near 41 kDa | Consistent with the predicted HOXA2 mass; confirm its identity |
| Stronger band near 41 kDa in nuclear extract | Consistent with HOXA2 nuclear localization |
| Faint band near 41 kDa in whole-cell lysate | Nuclear HOXA2 may be diluted in the sample |
| Little or no band in a cytosolic fraction | Consistent with HOXA2 nuclear localization |
💡Expected HOXA2 appearanceUniProt predicts HOXA2 at 41 kDa; no empirical band size is supplied, so a band near 41 kDa is a starting expectation that requires identity controls.
| UniProt predicted molecular weight | Provides a 41 kDa reference for band position |
| Calculated HOXA2 mass | 41,002 Da corresponds to approximately 41 kDa |
| Predicted mass reference | Does not establish the protein's apparent migration on a blot |
| Predicted 41 kDa size | Supports checking a band near 41 kDa with identity controls |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HOXA2 may be poorly represented in the sample | Check a nuclear extract and a suitable positive control |
| Band higher than expected | The supplied features do not establish a size-increasing modification | Compare with a positive control and verify antibody specificity |
| Band lower than expected | The supplied features do not establish precursor cleavage | Check sample integrity and verify band identity |
| Multiple bands | Only one isoform is listed; the extra bands are unexplained | Use specificity controls to identify the HOXA2 band |
| Weak or no signal | HOXA2 may be scarce in the tested sample | Check a suitable positive control and consider nuclear enrichment |
Experiment evidence
Western blot images showing HOXA2 band
A05671 · HOXA2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HOXA2 Western blot
🧪For positive controls for HOXA2 in Western blot, you can use no HPA-supported tissue or cell sample from the supplied data.
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.
⚠️Feasibility: HPA provides no tissue data, so a positive tissue control cannot be selected and a knockdown or KO is the practical negative control.
HPA tissue expression evidence for HOXA2
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 HOXA2 Western Blot Tips
Deeper troubleshooting and optimisation questions for HOXA2, answered from its protein features.
Where should the HOXA2 band appear?
Band shift · HOXA2 has a predicted mass of 41 kDa. Use that as a starting point when identifying a band; no observed Western blot band position was supplied.
Could a second band be a listed HOXA2 isoform?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. These features do not support assigning a second band to a documented isoform.
Does a different apparent mass establish a HOXA2 modification?
PTM · No. The supplied features list no modified residues or glycosylation sites, and feature data alone cannot explain an observed mass difference. Verify the band’s identity before assigning a cause.
No modified residues or glycosylation sites are listed. Their absence from the supplied features does not establish where an experimental band must migrate.
No modified residues or glycosylation sites are listed. Their absence from the supplied features does not establish where an experimental band must migrate.
Does this guide establish induction of HOXA2?
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 HOXA2?
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 A05671 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 HOXA2 Western blot signals be compared quantitatively?
Quantitation · Compare samples prepared from the same cellular fraction, because HOXA2 is nuclear. Use consistent loading and measure the same verified HOXA2 band in each lane.
Would nuclear samples help detect HOXA2?
Interpretation · HOXA2 is annotated as nuclear. A nuclear sample is therefore a reasonable choice when assessing its signal; keep sample preparation consistent across comparisons.
Do not assign them to an isoform or modification from these features alone: one isoform is listed, and no modified residues or glycosylation sites are listed. Check band identity before interpreting additional signals.
Do not assign them to an isoform or modification from these features alone: one isoform is listed, and no modified residues or glycosylation sites are listed. Check band identity before interpreting additional signals.
Boster reagents
HOXA2 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-A2 HOXA2 Antibody
Cat # A05671
A05671 is an anti-HOXA2 antibody listed for Western blotting with reported human and mouse reactivity. Its WB image uses extracts from various cell lines at a 1:1000 primary dilution; the caption does not identify the cell lines.
Which to pick: A05671 is the only listed HOXA2 antibody and has a WB image. Consider it for human or mouse samples based on reported reactivity, while noting that the image caption does not identify the tested cell lines.
Source: BosterBio HOXA2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43364.
- Human Protein Atlas. HOXA2 tissue expression.
- PMC6834402 — target-verified WB comparison
- PMC4499372 — target-verified WB comparison
- PMC3875777 — target-verified WB comparison