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- Table of Contents
LRP2 / Low-density lipoprotein receptor-related protein 2 · Western blot design guide
Design a Western Blot for LRP2
Real validated LRP2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LRP2 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
LRP2 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 | ~522 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Kidney (IHC candidate; verify WB) +2 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 LRP2 Western Blot Protocols
The M02463 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 293T cell lysate (catalog M02463) |
| Gel % | 4–12% gradient (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 | M02463; 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 LRP2 Western Blot Band Size?
LRP2 has a predicted precursor mass of 522 kDa; glycosylation and signal-peptide cleavage may affect migration, but no empirical band or demonstrated shift is supplied.
| Band near 522 kDa | consistent with the predicted full-length LRP2 mass; confirm identity with controls |
| Band above 522 kDa | N-linked glycosylation could affect apparent migration |
| Band slightly below 522 kDa | signal-peptide removal could reduce precursor mass slightly |
| Weak band in whole-cell lysate | apical membrane LRP2 may be poorly represented in the sample |
💡Expected LRP2 appearanceUniProt predicts a 522 kDa precursor; N-linked glycosylation and signal-peptide removal could affect migration, but no empirical band size is supplied, so confirm any candidate band with identity controls.
| Predicted precursor mass | sets a 522 kDa sequence-based reference |
| N-linked glycosylation at Asn159 | could increase apparent size; its individual effect is unknown |
| N-linked glycosylation at Asn178 | could increase apparent size; its individual effect is unknown |
| N-linked glycosylation at Asn299 | could increase apparent size; its individual effect is unknown |
| Signal peptide at residues 1–25 | cleavage makes the mature protein slightly smaller than the precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | apical membrane LRP2 may be poorly recovered | check a membrane-enriched fraction and a positive-control sample |
| Band higher than expected | N-linked glycosylation may alter migration | compare with a deglycosylated aliquot and confirm band identity |
| Band lower than expected | signal-peptide removal causes a small mass decrease; a larger difference remains unexplained | check antibody specificity and compare with a positive-control sample |
| Broad smear instead of sharp band | variation across N-linked glycosylation sites is possible | compare with a deglycosylated aliquot and check sample integrity |
| Multiple bands | glycosylation or signal-peptide processing may contribute, but distinct bands are unproven | compare bands with a positive-control sample and test antibody specificity |
Experiment evidence
Western blot images showing LRP2 band
M02463 · LRP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LRP2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LRP2 in Western blot, you can use kidney tissue, where HPA reports high expression.
Positive control: Kidney (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: Because LRP2 is an apical membrane protein, its signal may be diluted in whole-tissue lysate.
HPA tissue expression evidence for LRP2
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 |
|---|---|---|---|---|
| Kidney | proximal tubules (microvilli) | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Testis | Leydig 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LRP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for LRP2, answered from its protein features.
How should LRP2 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 listed LRP2 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to specific isoforms from these features alone. Check band identity using antibody specificity and the predicted 522 kDa mass as a reference.
Can glycosylation help identify an LRP2 band?
PTM · LRP2 has 40 listed N-linked glycosylation sites, including UniProt positions 159 and 4327. Compare untreated and deglycosylated aliquots with the same antibody if band identity is uncertain. A mobility change would support glycosylation; the site list alone does not predict its size.
UniProt lists phosphoserines at 4463, 4466, 4569, 4616 and 4653, plus phosphothreonine at 4632. These are full-length UniProt coordinates; check the numbering convention before comparing antibody or paper sites. The listed modifications do not establish a visible band shift.
UniProt lists phosphoserines at 4463, 4466, 4569, 4616 and 4653, plus phosphothreonine at 4632. These are full-length UniProt coordinates; check the numbering convention before comparing antibody or paper sites. The listed modifications do not establish a visible band shift.
Does this guide establish induction of LRP2?
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 LRP2 Western blot?
Transfer · LRP2 is a predicted 522 kDa, single-pass membrane protein. Choose and verify transfer conditions suitable for a protein this large; inspect the membrane and post-transfer gel to check whether high-mass protein transferred. The supplied features do not specify a validated transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02463 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 LRP2 bands be quantified?
Quantitation · Quantify the same well-resolved LRP2 band across samples within the assay’s linear range. LRP2 is listed at the apical cell membrane and in endosome-related locations, so keep sample preparation and fraction selection consistent. Do not combine bands of uncertain identity into one measurement.
Should LRP2 run at its predicted 522 kDa?
Interpretation · Use 522 kDa as the predicted mass, not an expected band position. LRP2 has a signal peptide at residues 1–25 and 40 listed N-linked glycosylation sites. These features may affect the mature protein’s mass or migration, but the supplied features do not establish an apparent band size or a visible shift.
The record lists a signal peptide at 1–25 but no propeptide or alternative sequence. Smaller bands cannot be assigned to a particular cleavage product or isoform from these features. Compare antibodies recognizing different LRP2 regions and assess whether the bands track with the intact protein.
The record lists a signal peptide at 1–25 but no propeptide or alternative sequence. Smaller bands cannot be assigned to a particular cleavage product or isoform from these features. Compare antibodies recognizing different LRP2 regions and assess whether the bands track with the intact protein.
Boster reagents
LRP2 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-Lrp2 / Megalin Rabbit Monoclonal Antibody
Cat # M02463
The catalog reports one anti-LRP2 antibody, M02463, with reported Human, Mouse, and Rat reactivity. Its Western blot image shows Lrp2 expression in 293T cell lysate; no additional WB validation is supplied.
Which to pick: M02463 is the only listed option. Its WB image uses 293T cell lysate, so consider whether that tested sample and its reported reactivity fit your experiment.
Source: BosterBio LRP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P98164.
- Human Protein Atlas. LRP2 tissue expression.
- PMC12084449 — target-verified WB comparison
- PMC11118422 — target-verified WB comparison
- PMC11668626 — target-verified WB comparison