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- Table of Contents
GAS1 / Growth arrest-specific protein 1 · Western blot design guide
Design a Western Blot for GAS1
Real validated GAS1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GAS1 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
GAS1 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 | ~35.7 kDa | |
| Observed band | ~40 kDa | |
| Gel | 5–20% (catalog A06815-2) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Oral mucosa (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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated GAS1 Western Blot Protocols
The A06815-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HEK293 (catalog A06815-2) |
| Gel % | 5–20% (catalog A06815-2) |
| Load | 30 ug; reducing conditions (catalog A06815-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06815-2) |
| Membrane | nitrocellulose membrane (catalog A06815-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06815-2) |
| Primary antibody | A06815-2 · 0.5 μg/mL (catalog A06815-2) |
| Primary incubation | overnight at 4°C (catalog A06815-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06815-2) |
| Secondary incubation | 1.5 hour at RT (catalog A06815-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06815-2) |
| Detection | ECL (catalog A06815-2) |
Section 2
What Is the Expected GAS1 Western Blot Band Size?
GAS1 is predicted at 35.7 kDa and observed near 40 kDa in reducing lysates; the cause of the difference is not established.
| Band near 40 kDa | Empirical GAS1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 35.7 kDa | Near the predicted precursor mass, not the validated band position |
| Band below the precursor position | May reflect signal-peptide or propeptide removal; identity needs confirmation |
| Band with altered mobility | N-linked glycosylation at Asn117 is possible; a visible shift is not established |
💡Expected GAS1 appearanceGAS1 has a predicted precursor mass of 35.7 kDa and an empirical band near 40 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted precursor mass | 35.7 kDa is the sequence-based reference, while the observed band is near 40 kDa |
| N-linked glycosylation at Asn117 | Could alter migration, but the size or visibility of any shift is unknown |
| Signal peptide at residues 1–39 | Cleavage shortens the precursor; apparent mature size is not supplied |
| Propeptide at residues 319–345 | Removal shortens the precursor; apparent mature size is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Asn117 glycosylation could affect migration, but its contribution to the 40 kDa band is unestablished | Compare with deglycosylated sample and confirm band identity |
| Band lower than expected | Signal-peptide or propeptide cleavage could produce a shorter form | Check antibody epitope coverage and compare precursor and processed samples |
| Multiple bands | Processing or Asn117 glycosylation could produce different forms; distinct bands are unproven | Compare deglycosylated and untreated samples and verify each band's identity |
| Weak or no signal | GAS1 is GPI-anchored at the cell membrane, which can affect recovery in lysate | Check membrane-protein recovery and use a positive control |
| Fragments below expected size | Signal-peptide or propeptide removal is annotated, but smaller fragments are not characterized | Check epitope coverage and assess sample proteolysis with identity controls |
Experiment evidence
Western blot images showing GAS1 band
A06815-2 · GAS1
A06815-1 · GAS1
A06815-1 · GAS1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GAS1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GAS1 in Western blot, you can use caudate tissue, which shows high HPA expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Oral mucosa (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because GAS1 is GPI-anchored at the cell membrane, membrane-enriched lysates may improve detection.
HPA tissue expression evidence for GAS1
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 |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced GAS1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GAS1, answered from its protein features.
How should GAS1 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.
Do annotated GAS1 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no annotated isoform difference to assign to multiple bands. Compare any additional bands with the annotated glycosylation site and processing regions without treating those features as proof of identity.
Which glycosylation site should I consider when interpreting the band?
PTM · UniProt lists one N-linked glycosylation site, Asn117, using UniProt sequence numbering. Glycosylation could affect migration, but the site annotation alone does not show that it causes a visible shift.
Does this guide establish induction of GAS1?
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 GAS1 Western blot?
Transfer · GAS1 is annotated at the cell membrane with a GPI anchor. The supplied features do not specify a transfer method, membrane, or duration. Check transfer efficiency for the approximately 40 kDa band when choosing conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06815-2 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 I quantify GAS1 across samples?
Quantitation · Quantify the same identified band across samples; the reported apparent band is about 40 kDa. GAS1 has an annotated glycosylation site and processing regions, so avoid combining distinct bands into one value unless their identities have been established.
Why does GAS1 run near 40 kDa instead of 35.7 kDa?
Interpretation · The observed band is about 40 kDa, while the predicted mass is 35.7 kDa. GAS1 has an N-linked glycosylation site at Asn117 and annotated processing regions. These features may affect the apparent mass, but they do not establish the cause of this difference.
UniProt annotates a signal peptide at residues 1..39 and a propeptide at 319..345, using its sequence numbering. When interpreting a band, check whether the antibody’s target region falls within either segment. These annotations alone do not identify a detected band as a particular processed form.
Consider the annotated Asn117 glycosylation site, signal peptide at 1..39, and propeptide at 319..345, all in UniProt numbering. These features suggest possibilities to investigate, but they do not establish the identity or cause of any unexpected band.
UniProt annotates a signal peptide at residues 1..39 and a propeptide at 319..345, using its sequence numbering. When interpreting a band, check whether the antibody’s target region falls within either segment. These annotations alone do not identify a detected band as a particular processed form.
Consider the annotated Asn117 glycosylation site, signal peptide at 1..39, and propeptide at 319..345, all in UniProt numbering. These features suggest possibilities to investigate, but they do not establish the identity or cause of any unexpected band.
Boster reagents
GAS1 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-GAS1 Antibody Picoband®
Cat # A06815-2
Real WB data
Anti-GAS1 Antibody
Cat # A06815-1
Two anti-GAS1 antibodies have supplied Western blot images. A06815-2 shows an approximately 40 kDa band in human HeLa and HEK293 lysates. A06815-1 shows MCF-7 lysate with a peptide-blocked lane. The supplied evidence does not establish broad sample validation.
Which to pick: For human HeLa or HEK293 lysates, A06815-2 has a documented 40 kDa band and detailed blot conditions. A06815-1 lists human and mouse reactivity, but its supplied WB image uses MCF-7 cells; no mouse WB example is provided.
Source: BosterBio GAS1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P54826.
- Human Protein Atlas. GAS1 tissue expression.
- PMC10583184 — target-verified WB comparison
- PMC10124827 — target-verified WB comparison
- PMC4832137 — target-verified WB comparison