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- Table of Contents
RELN / Reelin · Western blot design guide
Design a Western Blot for RELN
Real validated RELN Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RELN 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
RELN 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 | ~388.4 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Disulfide-linked oligomers | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated RELN Western Blot Protocols
The M02113 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M02113) |
| 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 | M02113; 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 RELN Western Blot Band Size?
Reelin's predicted full-length mass is 388.4 kDa; glycosylation, signal-peptide removal, isoforms, and disulfide-linked dimers could affect bands, but their migration effects are unverified.
| Band near 388.4 kDa | Compatible with the predicted full-length monomer; confirm identity with antibody controls |
| Band near twice the monomer size under non-reducing conditions | Compatible with a disulfide-linked homodimer |
| Slightly lower band | Could reflect signal-peptide removal, though a visible difference is not established |
| Little or no band in whole-cell lysate | Reelin is secreted into the extracellular space and matrix |
| Several bands at different positions | Isoforms 1, 2, and 3 are annotated, but distinct migration is not established |
💡Expected RELN appearanceUniProt predicts a 388.4 kDa full-length Reelin monomer; no empirical band size is supplied, so confirm any candidate band with antibody specificity and appropriate sample controls.
| Predicted full-length mass | 388.4 kDa is the sequence-based monomer reference, not a measured migration position |
| N-linked glycosylation at Asn140 and other annotated sites | May affect apparent migration; the size and visibility of any shift are not established |
| Disulfide-linked homodimers | Can yield a band near twice the monomer size under non-reducing or incompletely reducing conditions |
| Isoforms 1, 2, and 3 | May differ in size, but their masses and resolvable migration differences are not supplied |
| Signal peptide at residues 1–25 | Removal makes mature Reelin smaller than the full-length precursor; a resolvable shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Reelin is secreted into the extracellular space and matrix | Check conditioned medium or extracellular matrix material alongside lysate |
| Band higher than expected | Disulfide-linked homodimers may persist with incomplete reduction | Compare reducing and non-reducing preparations |
| Band lower than expected | Signal-peptide removal or isoform differences are possible; neither establishes this band's identity | Verify antibody specificity and compare an appropriate Reelin-positive control |
| Broad smear instead of sharp band | Variation in N-linked glycosylation is possible, but a smear is not established by site annotations | Compare matched samples before and after deglycosylation and verify antibody specificity |
| Multiple bands | Isoforms 1, 2, and 3 are annotated, but distinct bands are not established | Check band identity with a Reelin-positive control and an independent antibody |
Experiment evidence
Western blot images showing RELN band
M02113 · RELN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RELN Western blot
🧪For positive controls for RELN in Western blot, you can use no HPA-supported tissue or cell sample because none was supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside RELN.
⚠️Feasibility: RELN is secreted, so conditioned medium may be more useful than whole-cell lysate.
HPA tissue expression evidence for RELN
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced RELN Western Blot Tips
Deeper troubleshooting and optimisation questions for RELN, answered from its protein features.
How should RELN 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 RELN isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to the canonical 3460-residue sequence, isoform 2 lacks residues 3428–3429 and isoform 3 lacks residues 3428–3460. Check whether the antibody epitope falls in those regions before assigning a band to an isoform. These UniProt coordinates may differ from antibody or paper numbering.
First check antibody epitope coverage and the annotated isoform deletions: residues 3428–3429 in isoform 2 and 3428–3460 in isoform 3, using UniProt canonical numbering. Those deletions alone do not identify an observed band. No empirical RELN band sizes are supplied.
First check antibody epitope coverage and the annotated isoform deletions: residues 3428–3429 in isoform 2 and 3428–3460 in isoform 3, using UniProt canonical numbering. Those deletions alone do not identify an observed band. No empirical RELN band sizes are supplied.
How should glycosylation be considered when interpreting RELN bands?
PTM · RELN has 19 annotated N-linked glycosylation sites across its sequence. Compare samples under consistent preparation conditions and treat migration changes cautiously: the listed sites establish modification potential, but do not establish occupancy or the size of any band shift.
Does this guide establish induction of RELN?
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 RELN Western blot?
Transfer · RELN's predicted mass is 388.4 kDa, so verify that your transfer conditions recover high-mass protein. Check the membrane after transfer and retain a high-mass marker; the supplied features do not specify a transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02113 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.
Where should RELN be measured for quantitative comparison?
Quantitation · RELN is annotated as secreted to the extracellular space and extracellular matrix. Define whether you are measuring cell-associated, extracellular, or matrix-associated RELN, and compare the same fraction across samples. A cell lysate alone may not represent total RELN.
Why might RELN migrate differently from its predicted 388.4 kDa mass?
Interpretation · RELN has 19 annotated N-linked glycosylation sites and a signal peptide at residues 1–25. These features make the unmodified full-length sequence mass an imperfect migration reference. No observed band size is supplied, so neither feature establishes a visible shift or its magnitude.
RELN has 32 annotated disulfide bonds and is described as forming disulfide-linked homodimers within oligomers. Compare reducing and nonreducing preparations when evaluating high-mass bands, and document the preparation used. These features do not establish the identity of any particular band.
RELN has 32 annotated disulfide bonds and is described as forming disulfide-linked homodimers within oligomers. Compare reducing and nonreducing preparations when evaluating high-mass bands, and document the preparation used. These features do not establish the identity of any particular band.
Boster reagents
RELN 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-Reelin Rabbit Monoclonal Antibody
Cat # M02113
M02113 is an anti-Reelin rabbit monoclonal antibody listed for human and rat reactivity. Its Western blot image shows Reelin expression in HepG2 cell lysate; the supplied evidence does not document additional tested samples or conditions.
Which to pick: M02113 is the only listed RELN antibody. It has a Western blot image using HepG2 cell lysate; check whether that sample context and the listed human or rat reactivity fit your experiment.
Source: BosterBio RELN gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P78509.
- Human Protein Atlas. RELN tissue expression.
- PMC6667498 — target-verified WB comparison
- PMC11896430 — target-verified WB comparison
- PMC7318658 — target-verified WB comparison