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- Table of Contents
NLGN1 / Neuroligin-1 · Western blot design guide
Design a Western Blot for NLGN1
Source-linked NLGN1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NLGN1 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
NLGN1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~96.4 kDa | |
| Observed band | ~116 kDa | |
| Gel | 8% (catalog A04948-1) | |
| Positive control | Cerebral cortex (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 + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked NLGN1 Western Blot Protocol Options
The A04948-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human U251, rat C6 (catalog A04948-1) |
| Gel % | 8% (catalog A04948-1) |
| Load | 30 ug; reducing conditions (catalog A04948-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04948-1) |
| Membrane | nitrocellulose membrane (catalog A04948-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04948-1) |
| Primary antibody | A04948-1 · 0.5 μg/mL (catalog A04948-1) |
| Primary incubation | overnight at 4°C (catalog A04948-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04948-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04948-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04948-1) |
| Detection | ECL (catalog A04948-1) |
Section 2
What Is the Expected NLGN1 Western Blot Band Size?
NLGN1 is predicted at 96.4 kDa and observed near 116 kDa in reducing whole-cell lysates; the cause of the difference is not established.
| Band at ~116 kDa | Empirical NLGN1 band in reducing whole-cell lysates; its difference from the predicted mass is unexplained |
| Band near 96.4 kDa | Near the predicted mass of the full-length precursor; confirm its identity |
| Band below the precursor size | Signal-peptide cleavage could contribute to a smaller mature protein; identity requires confirmation |
| Multiple bands | Isoforms 1 and 2 or different glycosylation states are possible; distinct migration is unestablished |
| Broad smear | Variable N- or O-linked glycosylation is possible; the listed sites alone do not establish a smear |
💡Expected NLGN1 appearanceNLGN1 has a predicted precursor mass of 96.4 kDa and an empirical band at ~116 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.
| Predicted full-length mass | 96.4 kDa is the sequence-based precursor reference, while the empirical band is ~116 kDa |
| N-linked sites at Asn109, Asn323, Asn363, and Asn567 | Glycosylation could alter apparent migration; its contribution to the ~116 kDa band is unproven |
| O-linked sites at Ser703 and Ser706 | Glycosylation could alter apparent migration; site annotations do not establish a visible shift |
| Signal peptide at residues 1–45 | Cleavage can make mature protein smaller than the full-length precursor; no mature mass is supplied |
| Isoforms 1 and 2 | Alternative splicing may change size, but their masses and migration difference are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated NLGN1 may be poorly recovered | Check membrane extraction and test a membrane-enriched fraction |
| Band higher than expected | The ~116 kDa empirical band exceeds the 96.4 kDa predicted mass; the cause is unestablished | Compare with the ~116 kDa reference and confirm identity with an independent antibody or NLGN1-depleted sample |
| Band lower than expected | Signal-peptide cleavage or an alternative isoform is possible | Check antibody epitope coverage and confirm identity with an independent antibody or NLGN1-depleted sample |
| Broad smear instead of sharp band | Variable glycosylation is possible but not demonstrated | Compare matched samples with and without appropriate glycosidase treatment |
| Multiple bands | Isoforms or different glycosylation states are possible; distinct bands are not established | Use an independent antibody or NLGN1-depleted sample to identify specific bands |
| Weak or no signal | Membrane protein recovery or expression may be low in the sampled cells | Check lysate loading and extraction, then test an NLGN1-positive sample |
Experiment evidence
Western blot images showing NLGN1 band
A04948-1 · NLGN1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NLGN1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NLGN1 in Western blot, you can use cerebral cortex lysate, where HPA reports high expression.
Positive control: Cerebral cortex (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: As NLGN1 is a membrane protein, use a lysis method that recovers membrane proteins.
HPA tissue expression evidence for NLGN1
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 |
|---|---|---|---|---|
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Colon | glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NLGN1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NLGN1, answered from its protein features.
How should NLGN1 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 NLGN1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residues 165–204 in UniProt numbering. Check whether your antibody recognizes that region when interpreting bands. The deletion predicts a sequence difference, but does not establish where either isoform will migrate.
Which NLGN1 glycosylation sites matter when interpreting bands?
PTM · UniProt lists N-linked sites at Asn109, Asn323, Asn363 and Asn567, plus O-linked sites at Ser703 and Ser706. These are UniProt coordinates; antibody or paper numbering may differ. Glycosylation can complicate band interpretation, but the listed sites do not demonstrate a visible shift.
Does this guide establish induction of NLGN1?
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 NLGN1 Western blot?
Transfer · NLGN1 is a single-pass membrane protein with an observed band near 116 kDa. Check that this region transfers efficiently when optimizing your blot. The supplied UniProt features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04948-1 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 NLGN1 bands be quantified?
Quantitation · Use a consistent band region across samples and document which bands you include. NLGN1 has two isoforms, and its listed glycosylation sites may complicate band assignment. Confirm band identity before combining signals from bands at different positions.
Why might NLGN1 run near 116 kDa instead of 96.4 kDa?
Interpretation · The observed band is approximately 116 kDa, while the predicted mass is 96.4 kDa. UniProt lists a signal peptide at residues 1–45 and six glycosylation sites. These features may affect apparent migration, but their presence alone does not establish the cause of the difference.
Consider the isoform 2 deletion at residues 165–204, the signal peptide at residues 1–45, and the six listed glycosylation sites, all in UniProt numbering. NLGN1 also has three listed disulfide bonds. These features suggest possibilities to investigate; none identifies an unexpected band by itself.
Consider the isoform 2 deletion at residues 165–204, the signal peptide at residues 1–45, and the six listed glycosylation sites, all in UniProt numbering. NLGN1 also has three listed disulfide bonds. These features suggest possibilities to investigate; none identifies an unexpected band by itself.
Boster reagents
NLGN1 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-NLGN1 Antibody Picoband®
Cat # A04948-1
The catalog reports one anti-NLGN1 antibody for Western blot, A04948-1, with reported human and rat reactivity. Its WB image uses HeLa, U251, and rat C6 whole-cell lysates. The caption reports a band near 116 kDa, versus an expected 96 kDa; no independent validation is supplied.
Which to pick: A04948-1 is the only listed option and has a WB image from human HeLa and U251 and rat C6 lysates. Match your sample to those tested contexts and assess the reported 116 kDa band against the expected 96 kDa size.
Source: BosterBio NLGN1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8N2Q7.
- Human Protein Atlas. NLGN1 tissue expression.
- PMC7852195 — target-verified WB comparison
- PMC5762817 — target-verified WB comparison
- PMC10227424 — target-verified WB comparison