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LGR4 / Leucine-rich repeat-containing G-protein coupled receptor 4 · Western blot design guide
Design a Western Blot for LGR4
Real validated LGR4 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LGR4 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
LGR4 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 | ~104.5 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (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 | 2 isoform(s) |
Section 1
Real Curated LGR4 Western Blot Protocols
The A02134 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02134) |
| 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 | A02134; 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 LGR4 Western Blot Band Size?
LGR4’s predicted precursor is 104.5 kDa; glycosylation, signal-peptide cleavage and isoforms could affect migration, but no empirical band or visible shift is established.
| Band near 104.5 kDa | Consistent with the predicted LGR4 precursor mass; confirm identity with controls |
| Band above 104.5 kDa | Could reflect N-linked glycosylation at five annotated sites; migration is unverified |
| Band slightly below the precursor | Could reflect cleavage of the 1–24 signal peptide |
| Broad band or smear | Could reflect heterogeneous N-linked glycosylation, if present |
| Multiple bands | Could include isoforms 1 and 2 if their migration differs; verify band identity |
💡Expected LGR4 appearanceUniProt predicts a 104.5 kDa LGR4 precursor; signal-peptide cleavage, N-linked glycosylation and isoforms could affect migration, but no empirical band size is supplied, so confirm candidate bands with identity controls.
| Predicted precursor mass | 104.5 kDa is the sequence-based reference, not a measured band |
| N-linked glycosylation at Asn68, Asn199, Asn294, Asn314 and Asn505 | Could increase apparent size if these sites are glycosylated; the shift is unmeasured |
| Signal peptide at residues 1–24 | Its cleavage yields a smaller mature protein than the precursor |
| Splice isoforms 1 and 2 | Could differ in size, but their relative masses and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-localized LGR4 may be poorly recovered | Check a membrane-enriched fraction and a positive control |
| Band higher than expected | N-linked glycosylation could alter migration | Compare with a deglycosylated sample and confirm band identity |
| Band lower than expected | Signal-peptide cleavage could reduce the precursor mass | Check antibody epitope coverage and confirm band identity |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible | Compare with a deglycosylated sample and assess sample quality |
| Multiple bands | Isoforms 1 and 2 could differ in migration | Use isoform-aware controls or LGR4 depletion to identify bands |
Experiment evidence
Western blot images showing LGR4 band
A02134 · LGR4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LGR4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LGR4 in Western blot, you can use adrenal gland tissue, where HPA reports high expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, LGR4 may require effective membrane-protein extraction for a clear signal.
HPA tissue expression evidence for LGR4
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Low | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced LGR4 Western Blot Tips
Deeper troubleshooting and optimisation questions for LGR4, answered from its protein features.
How should LGR4 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 LGR4 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 62–85 of the canonical sequence, a region containing the annotated N-linked site at Asn68. Check whether the antibody epitope overlaps that region before interpreting a missing or additional band. These are canonical UniProt coordinates.
Which LGR4 glycosylation sites matter for band interpretation?
PTM · UniProt annotates N-linked sites at Asn68, Asn199, Asn294, Asn314, and Asn505 in the canonical sequence. Isoform 2 lacks the segment containing Asn68. Glycosylation may affect migration, but the site annotations alone cannot identify a band or predict its apparent mass.
UniProt lists phosphoserine at canonical residue 920. If assessing phosphorylation, use an assay that distinguishes phospho-Ser920 from total LGR4 and keep coordinate conventions explicit when comparing antibody or paper numbering. This annotation alone does not predict a detectable band shift or establish an induction condition.
UniProt lists phosphoserine at canonical residue 920. If assessing phosphorylation, use an assay that distinguishes phospho-Ser920 from total LGR4 and keep coordinate conventions explicit when comparing antibody or paper numbering. This annotation alone does not predict a detectable band shift or establish an induction condition.
Does this guide establish induction of LGR4?
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 LGR4 Western blot?
Transfer · LGR4 is a 951-residue multi-pass cell-membrane protein. Check recovery from the gel and retention on the membrane when optimizing transfer. The supplied features do not establish a specific transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02134 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 LGR4 bands?
Quantitation · Quantify the same consistently identified LGR4 band across samples. Document which isoform or band you measure, since isoform 2 lacks residues 62–85 and LGR4 has five annotated N-linked sites. The supplied record gives no observed band position to use as a fixed cutoff.
Should LGR4 run at its predicted 104.5 kDa?
Interpretation · 104.5 kDa is the predicted mass, not an established Western blot band position. LGR4 has a signal peptide at residues 1–24 and five annotated N-linked glycosylation sites. These features can affect apparent mass, but they do not establish the size or direction of a visible shift.
Check whether the antibody recognizes the region missing from isoform 2 (canonical residues 62–85), and consider the signal peptide at 1–24 and annotated N-linked sites when evaluating migration. The supplied features do not establish that any particular extra band represents an isoform, processing product, or glycosylation state.
Check whether the antibody recognizes the region missing from isoform 2 (canonical residues 62–85), and consider the signal peptide at 1–24 and annotated N-linked sites when evaluating migration. The supplied features do not establish that any particular extra band represents an isoform, processing product, or glycosylation state.
Boster reagents
LGR4 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-LGR4 Antibody
Cat # A02134
A02134 is an anti-LGR4 antibody listed for Western blot with stated human, mouse, and rat reactivity. A WB image is reported for extracts from various cell lines, but the supplied caption does not identify the lines or experimental conditions.
Which to pick: Only A02134 is listed. Choose it based on its stated reactivity and available WB image; check the full product details for sample identity and conditions before applying the example to your experiment.
Source: BosterBio LGR4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9BXB1.
- Human Protein Atlas. LGR4 tissue expression.
- PMC5777747 — target-verified WB comparison
- PMC9495020 — target-verified WB comparison
- PMC8422159 — target-verified WB comparison