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- Table of Contents
LRP6 / Low-density lipoprotein receptor-related protein 6 · Western blot design guide
Design a Western Blot for LRP6
Real validated LRP6 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LRP6 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
LRP6 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 | ~180.4 kDa | |
| Observed band | ~230 kDa | |
| Gel | 8% (catalog M00970) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Cervix (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 LRP6 Western Blot Protocols
The M00970 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Hela (catalog M00970) |
| Gel % | 8% (catalog M00970) |
| Load | 30 ug; reducing conditions (catalog M00970) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M00970) |
| Membrane | nitrocellulose membrane (catalog M00970) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M00970) |
| Primary antibody | M00970 · 1: 500 (catalog M00970) |
| Primary incubation | overnight at 4°C (catalog M00970) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M00970) |
| Secondary incubation | 1.5 hour at RT (catalog M00970) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M00970) |
| Detection | ECL (catalog M00970) |
Section 2
What Is the Expected LRP6 Western Blot Band Size?
LRP6 is predicted at 180.4 kDa and observed at ~230 kDa in reducing whole-cell blots; the cause of this difference is not established.
| Band at ~230 kDa | Empirical LRP6 band in reducing whole-cell lysates; predicted sequence mass is 180.4 kDa |
| Band near twice the monomer size under nonreducing conditions | May represent the disulfide-linked LRP6 homodimer |
| Band slightly below the full-length precursor | May reflect removal of the 19-residue signal peptide |
| Broad high-mass signal | Could reflect heterogeneous N-linked glycosylation or Wnt-associated oligomer aggregates; identity requires confirmation |
💡Expected LRP6 appearanceLRP6 has a predicted mass of 180.4 kDa, while reducing whole-cell blots show an empirical band at ~230 kDa; confirm its identity with antibody and sample controls because the cause of the difference is unestablished.
| Predicted LRP6 sequence mass | 180.4 kDa before accounting for apparent migration |
| N-linked glycosylation at Asn42, Asn81, Asn281, Asn433, Asn486, Asn692, Asn859 and Asn865 | May increase apparent mass; its contribution to the ~230 kDa band is unestablished |
| Disulfide-linked LRP6 homodimer | May yield a band near twice the monomer size without complete reduction |
| Signal peptide at residues 1–19 | Cleavage may make mature LRP6 slightly smaller than the precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated LRP6 may be poorly recovered | Check membrane protein extraction and include a positive-control lysate |
| Band higher than expected | Incomplete reduction may preserve the disulfide-linked homodimer | Compare fully reducing and nonreducing preparations |
| Band lower than expected | Signal peptide removal can slightly reduce precursor mass; a substantially lower band has no established identity here | Compare with a positive-control lysate and verify antibody specificity |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible but unconfirmed | Compare untreated and deglycosylated samples with an appropriate control |
| Multiple bands | Disulfide-linked dimers or Wnt-associated phosphorylated oligomer aggregates may contribute | Compare reducing conditions and use an independent LRP6 antibody |
| Weak or no signal | Membrane-associated LRP6 may be under-extracted or poorly transferred | Check extraction, high-mass transfer and a positive-control lysate |
Experiment evidence
Western blot images showing LRP6 band
M00970 · LRP6
M00970S1490 · LRP6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LRP6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LRP6 in Western blot, you can use adipose tissue lysate.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Cervix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside your samples.
⚠️Feasibility: LRP6 is a membrane protein, so use lysates that retain membrane proteins.
HPA tissue expression evidence for LRP6
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skin | fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LRP6 Western Blot Tips
Deeper troubleshooting and optimisation questions for LRP6, answered from its protein features.
How should LRP6 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.
Could annotated LRP6 isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. These records therefore provide no isoform-based explanation for multiple bands. Check band identity before assigning an additional band to an LRP6 isoform.
Which glycosylation sites are relevant when assessing LRP6 bands?
PTM · UniProt lists N-linked sites at Asn42, Asn81, Asn281, Asn433, Asn486, Asn692, Asn859, Asn865, Asn926 and Asn1039. A glycosylation-sensitive comparison could help assess band identity; the site list alone does not predict a visible shift.
Using the supplied UniProt coordinates: Ser1420 and Ser1430 are phosphorylated by CK1; Thr1479 is phosphorylated, with no enzyme specified; Ser1490 is phosphorylated by CDK14, GRK5 and GRK6; and Thr1493 is phosphorylated by CK1. Check the antibody's numbering convention before matching its target site to these coordinates.
LRP6 has listed glycosylation and phosphorylation sites, and its oligomer feature describes Wnt-associated aggregates. For comparisons, use the same sample preparation and quantify consistently defined LRP6 bands or regions. A change in one band's intensity alone need not represent a change in total LRP6.
Using the supplied UniProt coordinates: Ser1420 and Ser1430 are phosphorylated by CK1; Thr1479 is phosphorylated, with no enzyme specified; Ser1490 is phosphorylated by CDK14, GRK5 and GRK6; and Thr1493 is phosphorylated by CK1. Check the antibody's numbering convention before matching its target site to these coordinates.
LRP6 has listed glycosylation and phosphorylation sites, and its oligomer feature describes Wnt-associated aggregates. For comparisons, use the same sample preparation and quantify consistently defined LRP6 bands or regions. A change in one band's intensity alone need not represent a change in total LRP6.
Does this guide establish induction of LRP6?
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.
How should transfer be checked for LRP6?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M00970 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 LRP6 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 might LRP6 appear near 230 kDa instead of 180.4 kDa?
Interpretation · The observed band is approximately 230 kDa, while the predicted mass is 180.4 kDa. LRP6 has ten listed N-linked glycosylation sites, but these features alone cannot establish the cause or size of the apparent mass difference. Compare samples under the same gel conditions before assigning the band.
The supplied oligomer feature says LRP6 forms phosphorylated oligomer aggregates on Wnt signaling. Compare stimulated and control samples when interpreting higher-mass signal. This feature does not establish that a distinct shifted band will be visible or that total LRP6 expression increases.
LRP6 is described as a disulfide-linked homodimer and as forming phosphorylated oligomer aggregates on Wnt signaling. These features make oligomeric species relevant when assessing high-mass signal, but they do not identify any particular band. Compare reducing conditions and Wnt-treated controls where applicable.
The supplied oligomer feature says LRP6 forms phosphorylated oligomer aggregates on Wnt signaling. Compare stimulated and control samples when interpreting higher-mass signal. This feature does not establish that a distinct shifted band will be visible or that total LRP6 expression increases.
LRP6 is described as a disulfide-linked homodimer and as forming phosphorylated oligomer aggregates on Wnt signaling. These features make oligomeric species relevant when assessing high-mass signal, but they do not identify any particular band. Compare reducing conditions and Wnt-treated controls where applicable.
Boster reagents
LRP6 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-LRP6 Rabbit Monoclonal Antibody
Cat # M00970
Real WB data
Anti-Phospho-LRP6 (S1490) Rabbit Monoclonal Antibody
Cat # M00970S1490
Two rabbit monoclonal antibodies are listed for LRP6 Western blots: M00970 for total LRP6 and M00970S1490 for phospho-LRP6 (S1490). Both have WB images; the supplied captions show human cell lysates only. M00970’s observed band is approximately 230 kDa versus an expected 180 kDa.
Which to pick: Choose M00970 for total LRP6; its WB image uses human HepG2 and HeLa lysates. Choose M00970S1490 for S1490 phosphorylation; its image uses Calyculin-treated HeLa lysate. M00970 lists mouse and rat reactivity, but the supplied WB caption does not show those samples.
Source: BosterBio LRP6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75581.
- Human Protein Atlas. LRP6 tissue expression.
- PMC4575910 — target-verified WB comparison
- PMC7486456 — target-verified WB comparison
- PMC2882367 — target-verified WB comparison