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- Table of Contents
PIEZO1 / Piezo-type mechanosensitive ion channel component 1 · Western blot design guide
Design a Western Blot for PIEZO1
Real validated PIEZO1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PIEZO1 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
PIEZO1 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 | ~286.8 kDa | |
| Observed band | ~300 kDa | |
| Gel | 8% (catalog A01721-1) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PIEZO1 Western Blot Protocols
The A01721-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SiHa, human MCF-7 (catalog A01721-1) |
| Gel % | 8% (catalog A01721-1) |
| Load | 30 ug; reducing conditions (catalog A01721-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01721-1) |
| Membrane | nitrocellulose membrane (catalog A01721-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01721-1) |
| Primary antibody | A01721-1 · 1:1000 (catalog A01721-1) |
| Primary incubation | overnight at 4°C (catalog A01721-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01721-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01721-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01721-1) |
| Detection | ECL (catalog A01721-1) |
Section 2
What Is the Expected PIEZO1 Western Blot Band Size?
PIEZO1 is predicted at 286.8 kDa and observed near 300 kDa on reducing Western blots; the cause of the difference is not established.
| Band near 300 kDa | Empirical PIEZO1 band reported in reducing whole-cell lysates; confirm identity with controls |
| Band near 286.8 kDa | Migration near the predicted PIEZO1 sequence mass; confirm identity with controls |
| Diffuse signal near the main band | Possible variation in N-linked glycosylation at Asn295 or Asn2294; a visible effect is unproven |
| Weak or absent band in whole-cell lysate | Membrane-associated PIEZO1 may be poorly recovered during extraction |
💡Expected PIEZO1 appearancePIEZO1 has a predicted mass of 286.8 kDa, while antibody QC reports a band near 300 kDa in reducing whole-cell lysates; verify band identity with appropriate controls.
| Predicted PIEZO1 sequence mass | 286.8 kDa is the sequence-based reference; the reported band is near 300 kDa |
| N-linked glycosylation at Asn295 | May affect apparent migration; no site-specific shift is established |
| N-linked glycosylation at Asn2294 | May affect apparent migration; no site-specific shift is established |
| Disulfide bond between residues 2411 and 2415 | May affect migration under nonreducing conditions; no size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated PIEZO1 may be poorly extracted | Check membrane-enriched material and extraction conditions |
| Band higher than expected | A band near 300 kDa is reported, but its difference from 286.8 kDa is unexplained | Compare with a positive control and confirm PIEZO1 identity by depletion |
| Band lower than expected | Identity or sample integrity is uncertain | Check sample integrity and confirm the band by PIEZO1 depletion |
| Broad smear instead of sharp band | Variable glycosylation is possible but unproven | Compare untreated and deglycosylated samples with appropriate controls |
| Multiple bands | Band identities are uncertain; distinct isoform masses are not supplied | Compare bands against a positive control and PIEZO1-depleted sample |
| Weak or no signal | Membrane extraction or transfer of this large protein may be inefficient | Check recovery in the membrane fraction and transfer efficiency |
Experiment evidence
Western blot images showing PIEZO1 band
A01721-1 · PIEZO1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PIEZO1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PIEZO1 in Western blot, you can use cerebral cortex tissue, which HPA rates as high expression.
Positive control: Cerebral cortex (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: PIEZO1 is a multi-pass membrane protein, so membrane-enriched lysate may improve detection.
HPA tissue expression evidence for PIEZO1
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 |
|---|---|---|---|---|
| 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 PIEZO1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PIEZO1, answered from its protein features.
How should PIEZO1 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 multiple PIEZO1 isoform bands?
Isoforms · Only one isoform is supplied, with no alternative sequence listed. Do not assign additional bands to specific isoforms from this record alone; verify unexpected bands independently.
Could glycosylation shift the PIEZO1 band?
PTM · UniProt lists N-linked glycosylation at Asn295 and Asn2294, using its 2,521-residue sequence numbering. Glycosylation is a plausible source of mobility variation, but the site annotations do not demonstrate a visible shift. Compare treated and untreated samples if testing its contribution.
UniProt lists phosphothreonine at 734 and 1854, and phosphoserine at 758, 1391, 1396, 1636, and 1646. These are UniProt sequence coordinates; paper or antibody numbering may differ. The annotations do not establish a detectable band shift. Compare matched samples if testing phosphorylation-dependent mobility.
UniProt lists phosphothreonine at 734 and 1854, and phosphoserine at 758, 1391, 1396, 1636, and 1646. These are UniProt sequence coordinates; paper or antibody numbering may differ. The annotations do not establish a detectable band shift. Compare matched samples if testing phosphorylation-dependent mobility.
Does this guide establish induction of PIEZO1?
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 PIEZO1 Western blot?
Transfer · PIEZO1 is a 2,521-residue multi-pass membrane protein with an apparent band near 300 kDa. Use transfer conditions validated for proteins of that size, and check a high-molecular-weight marker and the post-transfer gel before interpreting a weak band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01721-1 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 PIEZO1 be quantified across samples?
Quantitation · Quantify the same verified ~300 kDa band across samples under consistent preparation and transfer conditions. PIEZO1 is a multi-pass membrane protein, so check transfer consistency before treating band intensity differences as abundance differences.
Why is the PIEZO1 band near 300 kDa instead of 286.8 kDa?
Interpretation · The supplied apparent band is ~300 kDa, while the predicted mass is 286.8 kDa. PIEZO1 has two annotated N-linked glycosylation sites and seven phosphorylated residues, but those features alone do not establish the cause of the difference. Use the ~300 kDa band as the empirical reference and verify its identity independently if needed.
PIEZO1 forms a homotrimer and has annotated glycosylation and phosphorylation sites, but these features do not identify any additional band by themselves. Compare unexpected bands with the ~300 kDa reference and verify their identity before assigning them to modifications, oligomers, or fragments.
PIEZO1 forms a homotrimer and has annotated glycosylation and phosphorylation sites, but these features do not identify any additional band by themselves. Compare unexpected bands with the ~300 kDa reference and verify their identity before assigning them to modifications, oligomers, or fragments.
Boster reagents
PIEZO1 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-Piezo1 (extracellular domain) Antibody
Cat # A01721-1
A01721-1 is listed for human and mouse PIEZO1. Its Western blot image uses human SiHa and MCF-7 whole cell lysates and shows a band near 300 kDa (expected 287 kDa). The supplied image does not demonstrate mouse samples.
Which to pick: A01721-1 is the only listed option and has a Western blot image from human cell lysates. For mouse samples, reactivity is listed, but no mouse Western blot image is supplied.
Source: BosterBio PIEZO1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92508.
- Human Protein Atlas. PIEZO1 tissue expression.
- PMC11737460 — target-verified WB comparison
- PMC11927149 — target-verified WB comparison
- PMC6685704 — target-verified WB comparison