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- Table of Contents
GBX2 / Homeobox protein GBX-2 · Western blot design guide
Design a Western Blot for GBX2
Source-linked GBX2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GBX2 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
GBX2 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 | ~37.3 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 GBX2 Western Blot Protocol Options
The A07270 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 | A07270; 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 GBX2 Western Blot Band Size?
GBX2 is predicted at 37.3 kDa; no empirical band is supplied, and its listed features do not demonstrate altered migration.
| Single band near 37.3 kDa | Consistent with predicted GBX2 mass; identity needs confirmation |
| Band near 37.3 kDa in nuclear extract | Consistent with GBX2's nuclear location |
| Band near 37.3 kDa enriched in nuclear extract | Consistent with nuclear localization of GBX2 |
| Little or no band in cytoplasmic extract | Consistent with GBX2's nuclear location |
💡Expected GBX2 appearanceUniProt predicts a 37.3 kDa GBX2 band, but no empirical band size is supplied; confirm a candidate band with ordinary identity controls.
| UniProt predicted molecular weight | Places full-length GBX2 near 37.3 kDa |
| Full-length predicted mass | Provides a 37.3 kDa reference for band position |
| Predicted mass in nuclear extract | Provides a 37.3 kDa reference for nuclear GBX2 |
| Predicted mass versus apparent migration | Does not establish where GBX2 migrates empirically |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear GBX2 may be poorly recovered | Check nuclear recovery and a positive control |
| Band higher than expected | No listed GBX2 feature establishes the shift | Check band identity with GBX2 depletion |
| Band lower than expected | No listed GBX2 feature establishes a smaller product | Check band identity with GBX2 depletion |
| Multiple bands | The single listed isoform does not explain distinct bands | Compare bands before and after GBX2 depletion |
| Weak or no signal | GBX2 abundance or nuclear recovery may be low | Check nuclear recovery and a positive control |
Experiment evidence
Western blot images showing GBX2 band
A07270 · GBX2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GBX2 Western blot
🧪For positive controls for GBX2 in Western blot, you can use a validated GBX2-positive sample once identified; the supplied HPA evidence names none.
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 suitable positive and negative samples cannot be confirmed.
HPA tissue expression evidence for GBX2
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 GBX2 Western Blot Tips
Deeper troubleshooting and optimisation questions for GBX2, answered from its protein features.
How should GBX2 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 GBX2 isoforms explain multiple bands?
Isoforms · The supplied features list one isoform and no alternative sequence. Multiple bands therefore cannot be assigned to annotated GBX2 isoforms from this evidence alone.
Do annotated modifications predict a GBX2 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features do not establish a modification-dependent shift or explain any difference between apparent and predicted mass.
Does this guide establish induction of GBX2?
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 GBX2?
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 A07270 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 GBX2 Western blot signal be quantified?
Quantitation · Compare GBX2 signal across samples prepared with the same fractionation method. Because GBX2 is annotated as nuclear, use comparable nuclear samples when drawing conclusions about nuclear abundance.
What molecular weight should guide GBX2 band identification?
Interpretation · The predicted mass is 37.3 kDa. No observed Western blot band position is supplied, so use 37.3 kDa as a reference rather than an expected apparent mass.
GBX2 is annotated as nuclear. Include a nuclear fraction when assessing its signal, and keep the fractionation method consistent across samples.
No signal peptide or propeptide is listed. The supplied features therefore provide no annotated processing event to assign to a smaller band.
Compare them with the 37.3 kDa predicted mass, but do not identify bands by migration alone. One isoform is listed, and no modified residues or glycosylation sites are annotated; these features do not establish the identity of an unexpected band.
GBX2 is annotated as nuclear. Include a nuclear fraction when assessing its signal, and keep the fractionation method consistent across samples.
No signal peptide or propeptide is listed. The supplied features therefore provide no annotated processing event to assign to a smaller band.
Compare them with the 37.3 kDa predicted mass, but do not identify bands by migration alone. One isoform is listed, and no modified residues or glycosylation sites are annotated; these features do not establish the identity of an unexpected band.
Boster reagents
GBX2 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 GBX-2 GBX2 Antibody
Cat # A07270
A07270 is an anti-GBX2 antibody listed for human and mouse reactivity. Its Western blot image uses K562 cell lysate, with primary antibody diluted 1:1000 overnight at 4°. The supplied evidence shows this test context only.
Which to pick: A07270 is the only listed GBX2 antibody and the only one with a Western blot image. Consider its listed reactivity for your samples; the K562 blot does not establish performance across other specimens.
Source: BosterBio GBX2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P52951.
- Human Protein Atlas. GBX2 tissue expression.
- PMC4355919 — target-verified WB comparison
- PMC3493575 — target-verified WB comparison