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- Table of Contents
GABPA / GA-binding protein alpha chain · Western blot design guide
Design a Western Blot for GABPA
Source-linked GABPA Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GABPA 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
GABPA 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 | ~51.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Lung (IHC candidate; verify WB) +4 more | |
| Negative control | Bone marrow (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked GABPA Western Blot Protocol Options
The A05350 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 | A05350; 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 GABPA Western Blot Band Size?
GABPA has a predicted monomer mass of 51.3 kDa; its phosphorylation and heterotetramer formation have no demonstrated effect on Western-blot migration here.
| Single band near 51.3 kDa | Consistent with the predicted GABPA monomer, pending band-identity controls |
| Band enriched in a nuclear fraction | Consistent with GABPA nuclear localization |
| Doublet near 51.3 kDa | Could reflect different phosphorylation states at Ser303; migration has not been demonstrated |
| Higher-mass species under native conditions | Could reflect the reported complex of two alpha and two beta subunits; its migration is unknown |
💡Expected GABPA appearanceGABPA has a predicted monomer mass of 51.3 kDa, but no empirical band size is supplied; confirm any candidate band with appropriate identity controls.
| UniProt predicted monomer mass | Sets a 51.3 kDa reference for the GABPA alpha chain |
| Phosphoserine at Ser303 | May affect mobility, but a visible shift is not established |
| Two-alpha, two-beta heterotetramer | May produce a higher-mass complex under native conditions; SDS migration is not established |
| Nuclear localization | Does not change molecular mass; may affect band recovery from a sample |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear GABPA may be poorly recovered | Check nuclear extraction and compare a nuclear fraction |
| Band higher than expected | Identity or migration of the species is unconfirmed | Compare denaturing and native conditions and verify band identity |
| Band lower than expected | Identity or protein integrity is unconfirmed | Verify band identity and check sample handling for degradation |
| Multiple bands | Different phosphorylation states are possible, but unresolved | Compare phosphatase-treated and untreated samples and verify band identity |
| Weak or no signal | Low recovery of nuclear GABPA or limited detection | Check nuclear fraction recovery and antibody performance |
| Fragments below expected size | Protein degradation is possible | Use fresh lysate with protease inhibitors and verify fragment identity |
Experiment evidence
Western blot images showing GABPA band
A05350 · GABPA
A05350-1 · GABPA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GABPA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GABPA in Western blot, you can use lung lysate, which has high HPA expression.
Positive control: Lung (IHC candidate; verify WB)
Negative control: Bone marrow (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As GABPA is nuclear, nuclear enrichment may improve detection.
HPA tissue expression evidence for GABPA
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 |
|---|---|---|---|---|
| Lung | alveolar cells type I | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GABPA Western Blot Tips
Deeper troubleshooting and optimisation questions for GABPA, answered from its protein features.
Where should the GABPA band appear?
Band shift · GABPA has a predicted mass of 51.3 kDa, but no observed band position is supplied. Use 51.3 kDa as a starting point, not a confirmed apparent mass.
Are multiple GABPA isoform bands expected?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning multiple bands to GABPA isoforms.
Can phosphorylation explain a shifted GABPA band?
PTM · UniProt lists phosphoserine at position 303 in its sequence numbering. This feature alone does not establish a visible shift or explain a difference between apparent and predicted mass.
Does this guide establish induction of GABPA?
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.
What transfer method to use for GABPA Western blot?
Transfer · Plan transfer for a protein predicted at 51.3 kDa and check transfer efficiency near that mass. The supplied features do not identify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05350 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 GABPA be quantified?
Quantitation · Measure the GABPA signal consistently across matched samples, accounting for its nuclear location. A phosphoserine 303 signal, if measured separately, should not be treated as total GABPA abundance.
How should GABPA's nuclear location affect sample selection?
Interpretation · GABPA is annotated as nuclear. If whole-cell signal is weak, examine a nuclear fraction and compare like fractions across samples.
GABPA forms a heterotetramer with two alpha and two beta subunits. That annotation alone does not identify an unexpected Western-blot band; verify its identity before attributing it to the complex.
GABPA forms a heterotetramer with two alpha and two beta subunits. That annotation alone does not identify an unexpected Western-blot band; verify its identity before attributing it to the complex.
Boster reagents
GABPA 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-GABP- alpha Monoclonal Antibody
Cat # A05350
Real WB data
Anti-GA-binding protein alpha chain GABPA Antibody
Cat # A05350-1
Two the supplier anti-GABPA antibodies have Western blot images. A05350 shows HeLa, A549, MCF-7, NIH/3T3, and SMMC-7721 lysates. A05350-1 reports various cell lines at 1:1000 but does not name them. No publication evidence is supplied.
Which to pick: Choose A05350 for an image using named cell lysates; its listed reactivity is Human and Mouse. Choose A05350-1 if you need listed Rat reactivity or a reported 1:1000 dilution. Both have WB images, but A05350-1's caption does not identify its cell lines.
Source: BosterBio GABPA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q06546.
- Human Protein Atlas. GABPA tissue expression.
- PMC5446731 — target-verified WB comparison
- PMC7244562 — target-verified WB comparison
- PMC9097325 — target-verified WB comparison