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- Table of Contents
GSC / Homeobox protein goosecoid · Western blot design guide
Design a Western Blot for GSC
Real validated GSC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GSC 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
GSC 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 | ~28.2 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
Real Curated GSC Western Blot Protocols
The M03084-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF7 cell lysate (catalog M03084-2) |
| 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 | M03084-2; 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 GSC Western Blot Band Size?
GSC is predicted at 28.2 kDa; no empirical band size is supplied, and the listed features do not demonstrate altered migration.
| Single band near 28.2 kDa | Consistent with the predicted GSC size; identity requires controls |
| Band enriched in nuclear fraction | Consistent with GSC nuclear localization |
| Little or no band in cytoplasmic fraction | Consistent with GSC nuclear localization |
| Multiple bands | Cannot be assigned to distinct annotated GSC isoforms |
💡Expected GSC appearanceGSC has a predicted mass of 28.2 kDa, but no empirical band size is supplied; use appropriate controls to establish the identity of any band near that size.
| UniProt predicted mass | Places the expected full-length band near 28.2 kDa |
| 28,150 Da sequence mass | Corresponds to approximately 28.2 kDa |
| 257-residue sequence | Underlies the full-length mass prediction |
| Single annotated isoform | Provides no basis for assigning separate isoform band sizes |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GSC may be difficult to detect without enriching its nuclear location | Check a nuclear fraction and a positive control |
| Band higher than expected | No listed GSC feature establishes a higher migrating species | Compare with a positive control and verify antibody specificity |
| Band lower than expected | No signal peptide or propeptide cleavage is annotated | Check band identity with an independent antibody |
| Multiple bands | The supplied record lists one isoform and does not explain extra bands | Use a positive control and an independent antibody to identify GSC |
| Weak or no signal | The sample may contain too little nuclear GSC to detect | Check a nuclear fraction and antibody performance with a positive control |
Experiment evidence
Western blot images showing GSC band
M03084-2 · GSC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GSC Western blot
🧪For positive controls for GSC in Western blot, you can use a confirmed GSC-positive nuclear lysate; the supplied HPA evidence identifies no specific tissue or cell line.
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 expression data, so specific positive and negative tissue controls cannot be selected.
HPA tissue expression evidence for GSC
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 GSC Western Blot Tips
Deeper troubleshooting and optimisation questions for GSC, answered from its protein features.
What band size should I expect for GSC?
Band shift · The predicted mass is 28.2 kDa. Use this as a reference, but no observed band size is supplied, so an apparent size or shift cannot be established from these features.
Could GSC isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands.
Do listed modifications predict a GSC band shift?
PTM · No modified residues or glycosylation sites are listed. These features do not support assigning a band shift to a specific modification.
Does this guide establish induction of GSC?
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 GSC?
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 M03084-2 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 GSC 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 relevant for detecting GSC?
Interpretation · GSC is listed as nuclear. Include a nuclear fraction when assessing whether a candidate band is consistent with its reported location.
Because GSC is listed as nuclear, compare equivalent nuclear fractions and account for differences in fraction loading when interpreting band intensity.
Compare candidate bands with the 28.2 kDa predicted mass and the reported nuclear location. The record supplies no observed band size, alternative sequence, or listed modification that identifies an unexpected band.
Because GSC is listed as nuclear, compare equivalent nuclear fractions and account for differences in fraction loading when interpreting band intensity.
Compare candidate bands with the 28.2 kDa predicted mass and the reported nuclear location. The record supplies no observed band size, alternative sequence, or listed modification that identifies an unexpected band.
Boster reagents
GSC 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-GSC Rabbit Monoclonal Antibody
Cat # M03084-2
The catalog reports one anti-GSC rabbit monoclonal antibody, M03084-2, with reported human reactivity. Its Western blot image shows GSC expression in MCF7 cell lysate. No publication evidence or broader sample validation is supplied.
Which to pick: M03084-2 is the only listed option for GSC. Its Western blot image uses MCF7 cell lysate; consider whether that sample context fits your experiment.
Source: BosterBio GSC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P56915.
- Human Protein Atlas. GSC tissue expression.
- PMC4208742 — target-verified WB comparison