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GABRA1 / Gamma-aminobutyric acid receptor subunit alpha-1 · Western blot design guide
Design a Western Blot for GABRA1
Real validated GABRA1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GABRA1 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
GABRA1 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 | ~51.8 kDa | |
| Observed band | ≈52 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +1 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated GABRA1 Western Blot Protocols
The PA1578 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat Brain, Rat Brain (catalog PA1578) |
| 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 | PA1578; 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 GABRA1 Western Blot Band Size?
GABRA1 is predicted at 51.8 kDa and observed at ≈52 kDa; the cause of their small difference is not established.
| Band at ≈52 kDa | Matches the empirical GABRA1 band; confirm identity with antibody and sample controls |
| Band above ≈52 kDa | N-linked glycosylation at Asn38 or Asn138 could affect migration, but a visible shift is unproven |
| Band below ≈52 kDa | Signal-peptide cleavage could lower mass relative to the precursor; confirm band identity |
| Weak band in whole-cell lysate | GABRA1 is a multi-pass membrane protein that may be underrepresented in the lysate |
💡Expected GABRA1 appearanceUniProt predicts 51.8 kDa, while antibody QC reports a band at ≈52 kDa; use antibody and sample controls to confirm its identity, since the supplied features do not establish a migration effect.
| UniProt predicted mass | The 51.8 kDa precursor mass is close to the observed ≈52 kDa band |
| N-linked glycosylation at Asn38 | Could affect apparent mass, but no visible shift is established |
| N-linked glycosylation at Asn138 | Could affect apparent mass, but no visible shift is established |
| Signal peptide at residues 1–27 | Cleavage would reduce mature protein mass relative to the precursor; its band position is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or no signal | A membrane protein may be poorly recovered or transferred | Check membrane protein recovery, transfer, and a positive-control lysate |
| No band in lysate | GABRA1 may be underrepresented in the sampled lysate | Check a membrane-enriched fraction and a positive-control lysate |
| Band higher than expected | N-linked glycosylation could alter migration | Compare with a validated deglycosylation control and confirm band identity |
| Band lower than expected | Signal-peptide cleavage could lower mass relative to the precursor | Confirm band identity with a second antibody or an appropriate control |
| Multiple bands | Different glycosylation or processing states are possible but unconfirmed | Check band identity with a second antibody and compare appropriate processing controls |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible but unconfirmed | Compare with a validated deglycosylation control and check sample quality |
Experiment evidence
Western blot images showing GABRA1 band
PA1578 · GABRA1
PA1578 · GABRA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GABRA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GABRA1 in Western blot, you can use cerebellum lysate, where HPA reports high expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, GABRA1 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for GABRA1
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 |
|---|---|---|---|---|
| Cerebellum | synaptic glomeruli - capsule | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GABRA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GABRA1, answered from its protein features.
How should GABRA1 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.
Should multiple GABRA1 bands be assigned to isoforms?
Isoforms · The supplied features list one isoform and no alternative sequence. Do not label additional bands as GABRA1 isoforms based on this record; verify their identity independently.
Which GABRA1 modifications matter when interpreting a blot?
PTM · UniProt lists N-linked glycosylation at Asn38 and Asn138, using UniProt coordinates. It lists no modified residues beyond those glycosylation sites. If comparing antibody or paper residue numbers, check the numbering convention before matching sites. Glycosylation site presence alone does not establish a visible band shift.
Does this guide establish induction of GABRA1?
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 GABRA1 Western blot?
Transfer · GABRA1 is a multipass membrane protein with an apparent band near 52 kDa. The supplied features do not specify a transfer method. Check transfer efficiency around 52 kDa in your membrane preparation and adjust conditions based on the result.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA1578 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 GABRA1 bands be quantified across samples?
Quantitation · GABRA1 is reported at the postsynaptic cell membrane, cell membrane, and cytoplasmic vesicle membrane. Use the same sample fraction and preparation across comparisons, quantify the same verified band near 52 kDa, and normalize consistently. A change in a membrane fraction need not represent the same change in total cellular GABRA1.
Why might the GABRA1 band differ from its predicted mass?
Interpretation · The predicted mass is 51.8 kDa, and the supplied observed band is approximately 52 kDa. GABRA1 has a signal peptide at residues 1–27 and N-linked glycosylation sites at Asn38 and Asn138, using UniProt coordinates. These features alone do not establish a visible shift or explain the apparent mass.
First verify which band represents GABRA1. The supplied record lists one isoform, a signal peptide at residues 1–27, N-linked glycosylation at Asn38 and Asn138, and a heteropentameric receptor. None of these features alone identifies an unexpected band or proves its cause. Keep UniProt numbering explicit when comparing site annotations.
First verify which band represents GABRA1. The supplied record lists one isoform, a signal peptide at residues 1–27, N-linked glycosylation at Asn38 and Asn138, and a heteropentameric receptor. None of these features alone identifies an unexpected band or proves its cause. Keep UniProt numbering explicit when comparing site annotations.
Boster reagents
GABRA1 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-GABA A Receptor alpha 1/GABRA1 Antibody Picoband®
Cat # PA1578
PA1578 is an anti-GABRA1 antibody listed for Western blot with human, mouse, and rat reactivity. Its supplied WB image shows two lanes of rat brain tissue lysate; the provided evidence does not show human or mouse WB samples.
Which to pick: PA1578 is the only listed option. Choose it if its stated reactivity fits your sample; the supplied WB image documents rat brain tissue lysate under the reported conditions.
Source: BosterBio GABRA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P14867.
- Human Protein Atlas. GABRA1 tissue expression.
- PMC9185876 — target-verified WB comparison
- PMC12247753 — target-verified WB comparison
- PMC8323041 — target-verified WB comparison