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- Table of Contents
LYVE1 / Lymphatic vessel endothelial hyaluronic acid receptor 1 · Western blot design guide
Design a Western Blot for LYVE1
Real validated LYVE1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LYVE1 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
LYVE1 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 | ~35.2 kDa | |
| Observed band | ~70 kDa | |
| Gel | 10% (catalog A04027-2) | |
| Positive control | Colon (IHC candidate; verify WB) +1 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated LYVE1 Western Blot Protocols
The A04027-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human HepG2, human HEL (catalog A04027-2) |
| Gel % | 10% (catalog A04027-2) |
| Load | 30 ug; reducing conditions (catalog A04027-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04027-2) |
| Membrane | nitrocellulose membrane (catalog A04027-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04027-2) |
| Primary antibody | A04027-2 · 1:1000 (catalog A04027-2) |
| Primary incubation | overnight at 4°C (catalog A04027-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog A04027-2) |
| Secondary incubation | 1.5 hour at RT (catalog A04027-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04027-2) |
| Detection | ECL (catalog A04027-2) |
Section 2
What Is the Expected LYVE1 Western Blot Band Size?
LYVE1 is predicted at 35.2 kDa, but a reducing-condition blot shows about 70 kDa; the cause of this difference is not established.
| Band near 70 kDa under reducing conditions | Empirical LYVE1 band; its difference from the predicted mass is unexplained |
| Band near 35 kDa | Consistent with the predicted 35.2 kDa precursor mass; confirm identity |
| Higher band under nonreducing conditions | May reflect disulfide-linked LYVE1 homodimer |
| Band below the precursor position | May reflect signal-peptide cleavage; confirm identity |
💡Expected LYVE1 appearanceLYVE1 has a predicted precursor mass of 35.2 kDa, while antibody QC reports a band near 70 kDa under reducing conditions; the cause of the difference is unestablished, so confirm band identity with controls.
| Predicted precursor mass | 35.2 kDa by sequence; the empirical reducing-condition band is near 70 kDa |
| N-linked glycosylation at Asn53 | May affect apparent migration; no shift is established for this site |
| N-linked glycosylation at Asn130 | May affect apparent migration; no shift is established for this site |
| Disulfide-linked homodimer | May produce a band near twice the monomer mass without complete reduction |
| Signal peptide at residues 1–19 | Cleavage makes the mature chain smaller than the precursor; its band position is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated LYVE1 may be poorly recovered | Check membrane extraction and a positive-control lysate |
| Band higher than expected | A disulfide-linked homodimer may persist with incomplete reduction | Compare reducing and nonreducing samples; confirm identity with an independent antibody or LYVE1 depletion |
| Band lower than expected | Signal-peptide cleavage may reduce precursor size | Compare with a positive control and confirm band identity |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible at Asn53 and Asn130 | Compare untreated and deglycosylated samples; confirm identity |
| Multiple bands | Precursor processing or incomplete reduction may contribute | Compare reducing conditions and confirm each band with an independent antibody or LYVE1 depletion |
| Weak or no signal | Membrane protein recovery may be insufficient | Check extraction and loading against a positive control |
Experiment evidence
Western blot images showing LYVE1 band
A04027-2 · LYVE1
M04027 · LYVE1
A04027-1 · LYVE1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LYVE1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LYVE1 in Western blot, you can use colon tissue, which HPA scores as a medium positive.
Positive control: Colon (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: As a membrane protein, LYVE1 may require efficient membrane-protein extraction for a clear signal.
HPA tissue expression evidence for LYVE1
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 |
|---|---|---|---|---|
| Colon | endothelial cells | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LYVE1 Western Blot Tips
Deeper troubleshooting and optimisation questions for LYVE1, answered from its protein features.
How should LYVE1 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.
Can bands identify LYVE1 isoforms?
Isoforms · Use the linked UniProt isoform annotations; migration alone does not identify an isoform. No additional isoform-specific band assignment is established here.
Which LYVE1 glycosylation sites should I consider when interpreting bands?
PTM · UniProt lists N-linked glycosylation at positions 53 and 130. These are UniProt sequence coordinates; antibody or paper numbering may differ. A glycosidase-treated sample can help assess whether glycosylation affects migration, but the listed sites do not establish a particular band shift.
Does this guide establish induction of LYVE1?
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 LYVE1 Western blot?
Transfer · LYVE1 is a single-pass membrane protein, and an apparent band near 70 kDa is reported. Select and verify a transfer setup that retains and transfers membrane proteins around that size; inspect the membrane and post-transfer gel to check recovery. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04027-2 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 LYVE1 across samples?
Quantitation · Use the same sample preparation and reducing conditions across samples because LYVE1 is membrane-associated and forms a disulfide-linked homodimer. Quantify a consistently identified band within the assay’s linear range, and record which band was measured if multiple bands appear.
Why might LYVE1 appear near 70 kDa instead of its predicted 35.2 kDa?
Interpretation · LYVE1 has N-linked glycosylation sites at UniProt positions 53 and 130 and forms a disulfide-linked homodimer. Either feature merits investigation, but the feature list alone does not establish the cause of the observed ~70 kDa band. Compare reducing and nonreducing samples and assess glycosylation experimentally.
LYVE1 is annotated as a disulfide-linked homodimer and has two disulfide bonds. Compare matched reducing and nonreducing samples when assessing a higher band. A change would support a role for disulfide linkage, but band position alone cannot identify the species.
Check whether bands change with reduction or glycosidase treatment: LYVE1 has a disulfide-linked homodimer and N-linked sites at UniProt positions 53 and 130. Its 1–19 signal peptide may also affect the mature chain’s mass. These features suggest tests, not definitive identities for individual bands.
LYVE1 is annotated as a disulfide-linked homodimer and has two disulfide bonds. Compare matched reducing and nonreducing samples when assessing a higher band. A change would support a role for disulfide linkage, but band position alone cannot identify the species.
Check whether bands change with reduction or glycosidase treatment: LYVE1 has a disulfide-linked homodimer and N-linked sites at UniProt positions 53 and 130. Its 1–19 signal peptide may also affect the mature chain’s mass. These features suggest tests, not definitive identities for individual bands.
Boster reagents
LYVE1 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-LYVE1 Antibody
Cat # A04027-2
Real WB data
Anti-LYVE1 Rabbit Monoclonal Antibody
Cat # M04027
Real WB data
Anti-LYVE1 Antibody
Cat # A04027-1
All three the supplier anti-LYVE1 antibodies have WB images. A04027-2 reports a ~70 kDa band and M04027 a ~29 kDa band, versus 35 kDa expected in both captions. A04027-1's MCF-7 caption gives no band size. The images alone do not establish band identity.
Which to pick: For rat samples, M04027 and A04027-1 list rat reactivity; A04027-2 lists human and mouse despite rat tissue lanes in its image. Match the tested sample: A04027-2 includes tissue lysates, M04027 includes human, rat, and mouse cell lysates, and A04027-1 shows MCF-7 lysate.
Source: BosterBio LYVE1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y5Y7.
- Human Protein Atlas. LYVE1 tissue expression.
- PMC5783450 — target-verified WB comparison
- PMC10560114 — target-verified WB comparison
- PMC13041691 — target-verified WB comparison