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GABARAPL1 / Gamma-aminobutyric acid receptor-associated protein-like 1 · Western blot design guide
Design a Western Blot for GABARAPL1
Real validated GABARAPL1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GABARAPL1 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
GABARAPL1 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 | ~14 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Testis (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Cleaved | |
| Caveat | Lipidation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated GABARAPL1 Western Blot Protocols
The A03382 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Mouse brain, (catalog A03382) |
| Gel % | 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 | 0.2 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A03382) |
| Primary antibody | A03382 · 1:600 (catalog A03382) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A03382) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A03382) |
Section 2
What Is the Expected GABARAPL1 Western Blot Band Size?
GABARAPL1 is predicted at 14 kDa; propeptide processing and two isoforms may affect migration, but no empirical band or visible shift is established.
| Band near 14 kDa | consistent with the predicted protein size; identity needs confirmation |
| Slightly lower band | could reflect processing of the residue 117 propeptide; a visible shift is unproven |
| Bands at different positions | could reflect isoforms 1 and 2; distinct migration is unproven |
| Stronger band in a membrane fraction | consistent with its lipid-anchored vesicle localization |
💡Expected GABARAPL1 appearanceThe predicted mass is 14 kDa, but no empirical band is supplied; propeptide processing and isoforms may affect migration, so confirm any candidate band with antibody and loss-of-signal controls.
| UniProt predicted mass | sets a 14 kDa reference for the protein |
| Residue 117 propeptide | processing could slightly reduce mass; a visible shift is unproven |
| Isoform 1 | may migrate differently from isoform 2; its mass is not supplied |
| Isoform 2 | may migrate differently from isoform 1; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | vesicle-associated protein may be poorly recovered | check extraction and a membrane-containing fraction |
| Band higher than expected | band identity or migration is unresolved | compare with a second antibody and a loss-of-signal control |
| Band lower than expected | residue 117 propeptide processing is possible | verify identity with a loss-of-signal control |
| Multiple bands | isoforms 1 and 2 are annotated, but their migration is unknown | verify each band with isoform-aware or loss-of-signal controls |
| Weak or no signal | vesicle-associated target may be underrepresented in the preparation | check loading and recovery of membrane-associated proteins |
| Fragments below expected size | degradation during sample preparation is possible | prepare fresh lysate with protease inhibitors and confirm band identity |
Experiment evidence
Western blot images showing GABARAPL1 band
A03382 · GABARAPL1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GABARAPL1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GABARAPL1 in Western blot, you can use testis lysate, the highest-scoring HPA tissue.
Positive control: Testis (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists adipose tissue as not detected, but confirm the negative result in your lysate.
HPA tissue expression evidence for GABARAPL1
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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GABARAPL1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GABARAPL1, answered from its protein features.
What should I check if extra GABARAPL1 bands appear?
Band shift · Check whether the antibody recognizes both listed isoforms, including isoform 2’s altered sequence at positions 97–117. Consider the listed processing at residue 117 and lipid-associated localization when comparing sample preparations. None of these features alone identifies an extra band; confirm its identity experimentally.
Could GABARAPL1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the canonical sequence at positions 97–117 is replaced by a different, longer sequence. Check which isoform the antibody recognizes before assigning bands; the supplied features do not establish their apparent sizes.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of GABARAPL1?
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 GABARAPL1 Western blot?
Transfer · For a protein predicted at 14 kDa, use transfer conditions that retain small proteins, such as a 0.2 µm membrane, and check whether protein passes through the membrane. Optimize against a size marker near 14 kDa; the UniProt features do not specify a transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03382 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 can GABARAPL1 be quantified consistently?
Quantitation · Use the same sample preparation and band definition across samples. GABARAPL1 is listed at autophagosomes and cytoplasmic vesicle membranes, with a lipid anchor, so extraction or fraction choice may affect recovery. If multiple bands appear, quantify them separately until their identities are established.
Should GABARAPL1 run exactly at its predicted 14 kDa?
Interpretation · Use 14 kDa as a starting point, not an exact band position. UniProt lists processing at residue 117, a lipid anchor, and an alternative sequence in isoform 2. These features alone do not establish a visible shift or explain any apparent mass difference.
UniProt marks residue 117 as a propeptide and places a lipid-anchored form at the cytoplasmic vesicle membrane. Consider processing and membrane association when interpreting bands, but do not assign a band to either form from position alone. The residue number follows the supplied UniProt canonical sequence.
UniProt marks residue 117 as a propeptide and places a lipid-anchored form at the cytoplasmic vesicle membrane. Consider processing and membrane association when interpreting bands, but do not assign a band to either form from position alone. The residue number follows the supplied UniProt canonical sequence.
Boster reagents
GABARAPL1 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-GABARAPL1 Antibody
Cat # A03382
A03382 is an anti-GABARAPL1 antibody listed for human, mouse and rat. Its supplied Western blot image documents mouse brain lysate at 25 µg per lane and a 1:600 primary dilution; the image does not establish performance in every listed species or tissue.
Which to pick: A03382 is the only listed option. For mouse brain Western blots, its supplied image provides a starting reference: 25 µg lysate per lane, 1:600 primary dilution and 180-second ECL exposure. For human, rat or other tissues, verify performance in your samples.
Source: BosterBio GABARAPL1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H0R8.
- Human Protein Atlas. GABARAPL1 tissue expression.
- PMC5650359 — target-verified WB comparison
- PMC12678799 — target-verified WB comparison
- PMC5593540 — target-verified WB comparison