This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
LRP5 / Low-density lipoprotein receptor-related protein 5 · Western blot design guide
Design a Western Blot for LRP5
Real validated LRP5 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LRP5 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
LRP5 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 | ~179.1 kDa | |
| Observed band | 179 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Epididymis (IHC candidate; verify WB) +4 more | |
| 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 LRP5 Western Blot Protocols
The PA2273 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat Liver at 40ug, Mouse Liver at 40ug, NIH at 40ug Predicted bind size: 179KD Observed bind size: 179KD (catalog PA2273) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | PA2273; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected LRP5 Western Blot Band Size?
LRP5 is predicted at 179.1 kDa and observed at 179 kDa; the supplied evidence does not establish a feature-specific migration shift.
| Band at 179 kDa | Empirical LRP5 band, close to its 179.1 kDa predicted mass |
| Band near twice the monomer mass under non-reducing conditions | Possible disulfide-linked LRP5 homodimer |
| Band slightly below the full-length precursor | Possible LRP5 after cleavage of its residues 1–31 signal peptide |
| Bands above the monomer under Wnt signaling conditions | Possible phosphorylated LRP5 oligomer aggregates, reported by similarity |
💡Expected LRP5 appearanceLRP5 is predicted at 179.1 kDa and observed at 179 kDa; verify band identity with appropriate controls because its listed modifications and processing do not establish a distinct migration shift.
| UniProt predicted mass | 179.1 kDa predicted; the empirical band is 179 kDa |
| N-linked glycosylation at Asn93, Asn138, Asn446, Asn499, Asn705 and Asn878 | May affect apparent mass, but the sites alone do not establish a visible shift |
| Disulfide-linked homodimer | May yield a band near twice the monomer mass if reduction is incomplete |
| Residues 1–31 signal peptide cleavage | Makes mature LRP5 smaller than the full-length precursor; a separate band is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane LRP5 may be poorly recovered during extraction | Check membrane protein recovery and include a positive lysate control |
| Band higher than expected | Incomplete reduction may retain disulfide-linked homodimers | Compare reducing and non-reducing samples |
| Band lower than expected | Signal peptide removal makes mature LRP5 smaller than its precursor | Compare with the 179 kDa empirical band and verify identity with an independent antibody |
| Multiple bands | Monomer and disulfide-linked homodimer may both be present | Compare reducing and non-reducing lanes and verify band identity |
| Weak or no signal | Membrane extraction or recovery may be inadequate | Check extraction efficiency and include a positive lysate control |
| Broad smear instead of sharp band | Glycosylation heterogeneity is possible but is not established by the listed sites | Compare glycosidase-treated and untreated samples and verify LRP5 identity |
Experiment evidence
Western blot images showing LRP5 band
PA2273 · LRP5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LRP5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LRP5 in Western blot, you can use epididymis tissue, where HPA reports high expression.
Positive control: Epididymis (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, LRP5 may be easier to detect in membrane-enriched lysates.
HPA tissue expression evidence for LRP5
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 |
|---|---|---|---|---|
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | 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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LRP5 Western Blot Tips
Deeper troubleshooting and optimisation questions for LRP5, answered from its protein features.
How should LRP5 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 isoforms explain multiple LRP5 bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore do not support assigning additional bands to annotated isoforms.
Which glycosylation sites should inform band interpretation?
PTM · UniProt lists N-linked asparagines at positions 93, 138, 446, 499, 705, and 878. These are UniProt sequence coordinates; antibody or paper numbering may differ. Compare any apparent mobility change with an appropriate control before attributing it to glycosylation.
Does this guide establish induction of LRP5?
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 LRP5 Western blot?
Transfer · LRP5 is a 179.1 kDa single-pass membrane protein. Use transfer conditions validated for proteins around 179 kDa and check that LRP5 leaves the gel and reaches the membrane. The supplied features do not identify one required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA2273 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 LRP5 signal be quantified across samples?
Quantitation · LRP5 is found at membranes and in the endoplasmic reticulum, and MESD chaperones it to the plasma membrane. Compare consistently prepared samples and use the same band definition across lanes; changes in recovery or localization can affect the measured signal.
Why might LRP5 migrate differently from 179.1 kDa?
Interpretation · The 179.1 kDa prediction comes from the sequence. LRP5 has a signal peptide at residues 1–31 and six annotated N-linked glycosylation sites. These features may affect the mature protein’s mass or migration, but they do not establish a visible shift. The supplied observed band is about 179 kDa.
UniProt describes phosphorylated LRP5 oligomer aggregates during Wnt signaling by similarity. A Wnt-dependent signal is plausible, but the supplied features do not establish a specific phosphosite, band shift, or change in LRP5 abundance. Compare matched samples under the same detection conditions.
LRP5 forms a disulfide-linked homodimer and has 21 annotated disulfide bonds. UniProt also describes phosphorylated oligomer aggregates during Wnt signaling by similarity. These features make higher-mass species worth investigating, but a band’s identity cannot be established from its position alone. Compare reducing conditions and matched controls.
UniProt describes phosphorylated LRP5 oligomer aggregates during Wnt signaling by similarity. A Wnt-dependent signal is plausible, but the supplied features do not establish a specific phosphosite, band shift, or change in LRP5 abundance. Compare matched samples under the same detection conditions.
LRP5 forms a disulfide-linked homodimer and has 21 annotated disulfide bonds. UniProt also describes phosphorylated oligomer aggregates during Wnt signaling by similarity. These features make higher-mass species worth investigating, but a band’s identity cannot be established from its position alone. Compare reducing conditions and matched controls.
Boster reagents
LRP5 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-LRP5 Antibody Picoband®
Cat # PA2273
The catalog reports PA2273 for LRP5 Western blotting, with stated Human, Mouse, and Rat reactivity. Its WB image shows a 179 kDa band in rat and mouse liver lysates and NIH whole cell lysate. Evidence here is limited to that product image.
Which to pick: PA2273 is the only listed option. Its WB image uses 40 µg lysate per lane and antibody at 0.5 µg/mL; use those conditions as a starting point for the reported sample types.
Source: BosterBio LRP5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75197.
- Human Protein Atlas. LRP5 tissue expression.
- PMC9097834 — target-verified WB comparison
- PMC3978879 — target-verified WB comparison
- PMC8831954 — target-verified WB comparison