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- Table of Contents
SEL1L / Protein sel-1 homolog 1 · Western blot design guide
Design a Western Blot for SEL1L
Real validated SEL1L Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SEL1L 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
SEL1L 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 | ~88.8 kDa | |
| Observed band | ~100 kDa | |
| Gel | 5–20% (catalog A03343-2) | |
| Positive control | Pancreas (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
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 SEL1L Western Blot Protocols
The A03343-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human U-87MG, rat pancreas (catalog A03343-2) |
| Gel % | 5–20% (catalog A03343-2) |
| Load | 30 ug; reducing conditions (catalog A03343-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03343-2) |
| Membrane | nitrocellulose membrane (catalog A03343-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03343-2) |
| Primary antibody | A03343-2 · 0.5 μg/mL (catalog A03343-2) |
| Primary incubation | overnight at 4°C (catalog A03343-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03343-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03343-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03343-2) |
| Detection | ECL (catalog A03343-2) |
Section 2
What Is the Expected SEL1L Western Blot Band Size?
SEL1L has an 88.8 kDa predicted precursor and an approximately 100 kDa observed band; the cause of their difference is not established.
| Band near 100 kDa | Empirical SEL1L signal in reducing lysates; confirm identity with appropriate controls |
| Band below 100 kDa | Signal peptide cleavage or an alternative isoform could contribute; identity requires confirmation |
| High-mass band under non-reducing conditions | A homodimer or homooligomer is possible by similarity; its survival during electrophoresis is unproven |
| Multiple bands | Isoforms 1 and 2 or other SEL1L forms are possible; distinct migration is unproven |
| Broad band or smear | Heterogeneity at five N-linked glycosylation sites is possible; a visible effect is unproven |
💡Expected SEL1L appearanceUniProt predicts an 88.8 kDa precursor, while reducing immunoblots show an approximately 100 kDa band; the cause of the difference is unproven, so confirm identity with a positive control and an independent antibody.
| UniProt predicted precursor mass | 88.8 kDa is the sequence-based reference, while the observed band is approximately 100 kDa |
| N-linked glycosylation at Asn195, Asn217, Asn272, Asn431 and Asn608 | Could alter apparent mass or band width; the migration effect is unmeasured |
| Signal peptide at residues 1–21 | Cleavage makes the mature chain smaller than the precursor; its band position is unmeasured |
| Homodimer and homooligomer formation by similarity | Could yield higher-mass species if complexes persist during electrophoresis |
| Isoforms 1 and 2 | May differ in size, but their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ER membrane protein may be poorly extracted | Check detergent extraction and run a positive control lysate |
| Band higher than expected | The approximately 100 kDa observed band exceeds the 88.8 kDa prediction for an unestablished reason; persistent oligomers could add higher bands | Check the marker and antibody specificity; compare reducing and non-reducing samples for higher bands |
| Band lower than expected | Signal peptide cleavage or an alternative isoform is possible | Verify identity with an independent antibody and compare matched samples |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible | Compare matched samples with and without deglycosylation and verify band identity |
| Multiple bands | Isoforms 1 and 2 or processed SEL1L forms are possible | Check antibody specificity and compare reducing conditions |
| Weak or no signal | ER membrane extraction or low recovery may limit detection | Check lysate preparation, loading and a positive control |
Experiment evidence
Western blot images showing SEL1L band
A03343-2 · SEL1L
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SEL1L Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SEL1L in Western blot, you can use pancreas tissue lysate, which has high HPA expression.
Positive control: Pancreas (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists low expression in several tissues but none as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for SEL1L
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 |
|---|---|---|---|---|
| Pancreas | exocrine glandular cells | High | Protein (IHC) | HPA → |
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Endometrium | cells in endometrial stroma | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced SEL1L Western Blot Tips
Deeper troubleshooting and optimisation questions for SEL1L, answered from its protein features.
How should SEL1L 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.
How could SEL1L isoforms affect band interpretation?
Isoforms · Isoform 2 replaces GYRY at UniProt residues 298–301 with VSRL and lacks residues 302–794. It may yield a shorter band than isoform 1. Check whether the antibody epitope lies in a region retained by isoform 2 before assigning a band.
Possibly. UniProt lists isoform 2 with residues 302–794 missing and residues 298–301 changed. A lower band is compatible with a shorter sequence, but size alone cannot identify it. An antibody recognizing the retained portion of the protein could help assess that possibility.
Possibly. UniProt lists isoform 2 with residues 302–794 missing and residues 298–301 changed. A lower band is compatible with a shorter sequence, but size alone cannot identify it. An antibody recognizing the retained portion of the protein could help assess that possibility.
Which glycosylation sites matter when assessing SEL1L mobility?
PTM · UniProt lists N-linked glycosylation at Asn195, Asn217, Asn272, Asn431, and Asn608. Compare untreated and deglycosylated samples if testing whether glycans affect mobility. Site annotations alone do not establish a visible shift.
UniProt annotates phosphoserine at residue 63. A phosphorylation-sensitive comparison may help assess a suspected band difference, but this annotation does not establish that phosphorylation produces a resolvable shift. Residue 63 uses the supplied UniProt sequence numbering.
UniProt annotates phosphoserine at residue 63. A phosphorylation-sensitive comparison may help assess a suspected band difference, but this annotation does not establish that phosphorylation produces a resolvable shift. Residue 63 uses the supplied UniProt sequence numbering.
Does this guide establish induction of SEL1L?
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 SEL1L Western blot?
Transfer · SEL1L is a single-pass ER membrane protein, with an observed band near 100 kDa. A wet transfer is a practical starting point; check the membrane and post-transfer gel around 100 kDa to confirm transfer before interpreting a weak band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03343-2 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 SEL1L bands be quantified?
Quantitation · Quantify the same assigned SEL1L band across samples. The observed band is near 100 kDa, and isoform 2 has a large sequence deletion, so assess distinct bands separately rather than combining them without an isoform assignment.
Why might SEL1L appear near 100 kDa instead of 88.8 kDa?
Interpretation · The supplied blot observation is approximately 100 kDa, while the predicted mass is 88.8 kDa. SEL1L has a 1–21 signal peptide and five annotated N-linked glycosylation sites. These features are relevant to the mass comparison, but they do not establish the cause of the observed difference.
UniProt describes SEL1L as a homodimer and homooligomer by similarity and lists several interaction partners. Those annotations make complexes worth considering, but they do not identify a particular higher band. Compare samples under appropriately denaturing conditions before assigning one to an oligomer.
UniProt describes SEL1L as a homodimer and homooligomer by similarity and lists several interaction partners. Those annotations make complexes worth considering, but they do not identify a particular higher band. Compare samples under appropriately denaturing conditions before assigning one to an oligomer.
Boster reagents
SEL1L 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-SEL1L Antibody Picoband®
Cat # A03343-2
The catalog reports one anti-SEL1L antibody, A03343-2, with a Western blot image of human cell and rat and mouse pancreas lysates. The reported band is approximately 100 kDa versus an expected 89 kDa; the supplied evidence does not resolve this difference.
Which to pick: A03343-2 is the only listed option. Its stated reactivity covers human, mouse, and rat, and its WB image shows human HepG2 and U-87MG cells plus rat and mouse pancreas tissue. Match your sample to these tested contexts where possible.
Source: BosterBio SEL1L gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UBV2.
- Human Protein Atlas. SEL1L tissue expression.
- PMC12949252 — target-verified WB comparison
- PMC10873344 — target-verified WB comparison
- PMC10786691 — target-verified WB comparison