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- Table of Contents
LY6E / Lymphocyte antigen 6E · Western blot design guide
Design a Western Blot for LY6E
Source-linked LY6E Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LY6E 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
LY6E 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 | ~13.5 kDa | |
| Observed band | ≈14 kDa | |
| Gel | 5–20% (catalog A09496-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked LY6E Western Blot Protocol Options
The A09496-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549 (catalog A09496-2) |
| Gel % | 5–20% (catalog A09496-2) |
| Load | 30 ug; reducing conditions (catalog A09496-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A09496-2) |
| Membrane | nitrocellulose membrane (catalog A09496-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A09496-2) |
| Primary antibody | A09496-2 · 0.5 μg/mL (catalog A09496-2) |
| Primary incubation | overnight at 4°C (catalog A09496-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A09496-2) |
| Secondary incubation | 1.5 hour at RT (catalog A09496-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A09496-2) |
| Detection | ECL (catalog A09496-2) |
Section 2
What Is the Expected LY6E Western Blot Band Size?
LY6E is predicted at 13.5 kDa and observed near 14 kDa; the cause of that small difference is not established.
| Band near 14 kDa | Matches the reported LY6E band, close to the 13.5 kDa precursor prediction |
| Band below 14 kDa | Could reflect signal peptide or propeptide cleavage; identity needs confirmation |
| Band above 14 kDa | Could reflect N-linked glycosylation at Asn99; a visible shift is unproven |
| Little or no band in a membrane-depleted fraction | Consistent with GPI-anchored cell membrane localization |
💡Expected LY6E appearanceLY6E has a predicted precursor mass of 13.5 kDa and a reported band near 14 kDa; confirm band identity with an appropriate antibody control because the supplied features do not establish its processed or glycosylated state.
| UniProt predicted precursor mass | 13.5 kDa provides the sequence-based reference for the reported band near 14 kDa |
| N-linked glycosylation at Asn99 | May affect apparent size if occupied; no shift magnitude is established |
| Signal peptide at residues 1–20 | Cleavage reduces protein length, but mature migration is not supplied |
| Propeptide at residues 102–131 | Cleavage reduces protein length, but mature migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GPI-anchored membrane protein may be poorly recovered | Check membrane recovery and compare with a validated positive lysate |
| Band higher than expected | Asn99 glycosylation is possible, but the cause of a shift is unproven | Compare matched samples with and without N-glycan removal |
| Band lower than expected | Signal peptide or propeptide cleavage may change length | Confirm identity with an independent LY6E antibody or other band-identity control |
| Multiple bands | Processing or Asn99 glycosylation may contribute, but distinct bands are not established | Check band identity and compare matched samples after N-glycan removal |
| Weak or no signal | Membrane recovery may be inadequate for GPI-anchored LY6E | Check extraction and include a validated positive lysate |
| Fragments below expected size | Processing is annotated, but fragment masses are not supplied | Check sample integrity and verify smaller bands with an independent LY6E antibody |
Experiment evidence
Western blot images showing LY6E band
A09496-2 · LY6E
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LY6E Western blot
🧪For positive controls for LY6E in Western blot, you can use no evidence-backed positive sample from the supplied HPA data.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: HPA data are unavailable, so tissue controls cannot be validated; a knockdown or KO line can provide a negative control.
HPA tissue expression evidence for LY6E
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced LY6E Western Blot Tips
Deeper troubleshooting and optimisation questions for LY6E, answered from its protein features.
How should LY6E 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.
Are alternative LY6E isoforms expected?
Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign additional bands to annotated isoforms on this evidence alone.
Could glycosylation change LY6E migration?
PTM · UniProt lists one N-linked glycosylation site at Asn99, numbered in the full-length sequence. Glycosylation could affect migration, but the site annotation alone does not establish a detectable shift. Compare treated and untreated samples if assigning a shifted band to glycosylation.
Does this guide establish induction of LY6E?
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 LY6E Western blot?
Transfer · LY6E has a predicted mass of 13.5 kDa and an observed band near 14 kDa. Use transfer conditions that retain proteins in this size range, and check transfer efficiency around 14 kDa. The supplied features do not establish a specific membrane or transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09496-2 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 LY6E bands be quantified?
Quantitation · Quantify a consistently defined band near the observed 14 kDa position across samples. Keep any additional bands separate until their identity is established; the listed processing and glycosylation features do not show that those bands are LY6E.
Why is the LY6E band near its predicted mass?
Interpretation · The observed band is approximately 14 kDa, close to the 13.5 kDa predicted mass. LY6E has a signal peptide (UniProt residues 1..20), a propeptide (102..131), and an N-linked glycosylation site at Asn99. These features alone do not establish a visible shift or explain the small difference.
Consider the signal peptide at UniProt residues 1..20, propeptide at 102..131, N-linked site at Asn99, and GPI anchoring when evaluating band identity. These full-length sequence coordinates may differ from antibody or paper numbering. None of the annotations alone identifies an unexpected band or proves a visible shift.
Consider the signal peptide at UniProt residues 1..20, propeptide at 102..131, N-linked site at Asn99, and GPI anchoring when evaluating band identity. These full-length sequence coordinates may differ from antibody or paper numbering. None of the annotations alone identifies an unexpected band or proves a visible shift.
Boster reagents
LY6E 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-LY6E/SCA-2 Antibody Picoband®
Cat # A09496-2
The catalog reports one anti-LY6E antibody, A09496-2, with reported human reactivity. Its Western blot image shows a band near 14 kDa in human A549 whole-cell lysate under reducing conditions. The supplied evidence covers this sample and assay only.
Which to pick: A09496-2 is the only listed option. It has a Western blot image from human A549 whole-cell lysate, using 30 µg of sample and 0.5 µg/mL primary antibody. Use that reported context to judge its fit for your experiment.
Source: BosterBio LY6E gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16553.
- Human Protein Atlas. LY6E tissue expression.
- PMC7459569 — target-verified WB comparison
- PMC13570520 — target-verified WB comparison
- PMC4910623 — target-verified WB comparison