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- Table of Contents
AQP8 / Aquaporin-8 · Western blot design guide
Design a Western Blot for AQP8
Source-linked AQP8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AQP8 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
AQP8 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 | ~27.4 kDa | |
| Observed band | ~27 kDa | |
| Gel | 5–20% (catalog PA1511) | |
| Positive control | Pancreas (IHC candidate; verify WB) +2 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked AQP8 Western Blot Protocol Options
The PA1511 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A431 (catalog PA1511) |
| Gel % | 5–20% (catalog PA1511) |
| Load | 50ug; reducing conditions (catalog PA1511) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog PA1511) |
| Membrane | Nitrocellulose membrane (catalog PA1511) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog PA1511) |
| Primary antibody | PA1511 · 0.5 μg/mL (catalog PA1511) |
| Primary incubation | overnight at 4°C (catalog PA1511) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog PA1511) |
| Secondary incubation | 1.5 hour at RT (catalog PA1511) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1511) |
| Detection | ECL (catalog PA1511) |
Section 2
What Is the Expected AQP8 Western Blot Band Size?
AQP8 is predicted at 27.4 kDa and observed at ~27 kDa; its N-linked sites do not establish a visible migration shift.
| Band at ~27 kDa | Observed AQP8 band, close to its 27.4 kDa predicted mass |
| Band above ~27 kDa | Could reflect N-linked glycosylation at N59 or N139; a shift is not established |
| Broad signal around the main band | Could reflect variable N-linked glycosylation; the sites alone do not establish a smear |
| Little signal in a soluble fraction | AQP8 is a multi-pass membrane protein |
💡Expected AQP8 appearanceAQP8 has a predicted mass of 27.4 kDa and an observed band at ~27 kDa; confirm band identity with appropriate antibody and sample controls.
| Predicted AQP8 mass | 27.4 kDa predicted; the observed band is ~27 kDa |
| N-linked glycosylation site N59 | Occupancy could affect apparent size, but no shift is established |
| N-linked glycosylation site N139 | Occupancy could affect apparent size, but no shift is established |
| Combined occupancy of N59 and N139 | Could alter apparent migration; the extent is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-localized AQP8 may be poorly recovered | Check membrane extraction and include a positive lysate control |
| Band higher than expected | N-linked glycosylation is possible, but the shift is unproven | Compare with a validated AQP8 control and test glycosidase sensitivity |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unconfirmed | Check glycosidase sensitivity and compare with a positive control |
| Multiple bands | Glycosylation states or nonspecific antibody binding may contribute | Compare with a validated AQP8 control and check antibody specificity |
| Weak or no signal | Recovery of multi-pass membrane AQP8 may be low | Check membrane protein recovery and use a positive lysate control |
Experiment evidence
Western blot images showing AQP8 band
PA1511 · AQP8
PA1511 · AQP8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AQP8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AQP8 in Western blot, you can use pancreas tissue, which has high HPA expression.
Positive control: Pancreas (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: Because AQP8 is a multi-pass membrane protein, membrane-enriched lysate may improve detection.
HPA tissue expression evidence for AQP8
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 |
|---|---|---|---|---|
| Pancreas | exocrine glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Rectum | glandular cells | 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 → |
| 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 AQP8 Western Blot Tips
Deeper troubleshooting and optimisation questions for AQP8, answered from its protein features.
How should AQP8 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 the supplied features support an AQP8 isoform explanation?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no specific alternative isoform to assign to an unexpected band.
Could glycosylation affect AQP8 band migration?
PTM · UniProt lists N-linked glycosylation sites at Asn59 and Asn139. These sites make glycosylation relevant when assessing additional bands, but their presence alone does not establish occupancy or a visible shift. The positions are UniProt coordinates and may differ from antibody or paper numbering.
UniProt lists two modifications at Cys53: cysteine persulfide and cysteine sulfenic acid (-SOH). Cys53 uses UniProt numbering, which may differ from paper or antibody numbering. These features alone do not show that either modification produces a detectable band shift.
UniProt lists two modifications at Cys53: cysteine persulfide and cysteine sulfenic acid (-SOH). Cys53 uses UniProt numbering, which may differ from paper or antibody numbering. These features alone do not show that either modification produces a detectable band shift.
Does this guide establish induction of AQP8?
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 AQP8 Western blot?
Transfer · AQP8 is a 261-residue, multi-pass membrane protein with a predicted mass of 27.4 kDa. Check that the approximately 27 kDa protein transfers consistently across samples. The supplied features do not specify a transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA1511 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 AQP8 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.
Why is AQP8 observed near 27 kDa?
Interpretation · AQP8 has a predicted mass of 27.4 kDa, close to the reported apparent band near 27 kDa. That agreement is consistent with the prediction; the supplied features do not establish what determines its exact migration.
AQP8 is listed at the cell membrane and mitochondrial inner membrane, with an apical cell membrane annotation. Compare like sample fractions and preparation methods when quantifying the approximately 27 kDa band; a change in fraction can change the amount of AQP8 sampled.
Start with the reported band near 27 kDa. Asn59 and Asn139 glycosylation sites and Cys53 modification annotations are relevant possibilities to investigate, but they do not identify an unexpected band by themselves. The supplied record lists no alternative sequence to support a specific isoform assignment.
AQP8 is listed at the cell membrane and mitochondrial inner membrane, with an apical cell membrane annotation. Compare like sample fractions and preparation methods when quantifying the approximately 27 kDa band; a change in fraction can change the amount of AQP8 sampled.
Start with the reported band near 27 kDa. Asn59 and Asn139 glycosylation sites and Cys53 modification annotations are relevant possibilities to investigate, but they do not identify an unexpected band by themselves. The supplied record lists no alternative sequence to support a specific isoform assignment.
Boster reagents
AQP8 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-Aquaporin 8/AQP8 Antibody Picoband®
Cat # PA1511
PA1511 is listed for human, mouse, and rat AQP8. Its Western blot image shows a band near the expected 27 kDa in human A431 whole cell lysate. The supplied evidence does not show mouse or rat Western blots.
Which to pick: PA1511 is the only listed antibody. It has a Western blot image using human A431 lysate; consider your sample species and confirm performance in your own material.
Source: BosterBio AQP8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O94778.
- Human Protein Atlas. AQP8 tissue expression.
- PMC8673491 — target-verified WB comparison
- PMC10253139 — target-verified WB comparison
- PMC9196045 — target-verified WB comparison