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SNX1 / Sorting nexin-1 · Western blot design guide
Design a Western Blot for SNX1
Source-linked SNX1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SNX1 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
SNX1 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 | ~59.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cervix (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SNX1 Western Blot Protocol Options
The M02692-1 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 M02692-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02692-1; 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 SNX1 Western Blot Band Size?
SNX1 is predicted at 59.1 kDa; isoforms, phosphorylation and self-association could affect migration, but no empirical band or visible shift is established.
| Band near 59.1 kDa | Consistent with the predicted SNX1 monomer; confirm its identity with a loss-of-signal control. |
| Additional band at a different size | Could reflect isoforms 1, 1A or 3; their migration differences are unknown. |
| Close doublet | Could reflect phosphorylation-dependent mobility, which is not established for this blot. |
| Band near twice the monomer size | Could reflect SNX1 self-association if a homodimer survives sample preparation. |
💡Expected SNX1 appearanceUniProt predicts a 59.1 kDa SNX1 monomer, but no empirical band size is supplied; isoforms, phosphorylation and self-association warrant band-identity controls before assigning other signals.
| UniProt predicted monomer mass | Sets a 59.1 kDa reference, not a measured migration position. |
| Isoform 1 | May differ in size from isoforms 1A and 3; the direction is unspecified. |
| Isoform 1A | May differ in size from isoforms 1 and 3; the direction is unspecified. |
| Isoform 3 | May differ in size from isoforms 1 and 1A; the direction is unspecified. |
| SNX1 homodimer formation | Could yield a higher band if the complex survives sample preparation; SDS gel migration is unverified. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated SNX1 may be poorly recovered. | Check extraction conditions and test a membrane-enriched fraction. |
| Band higher than expected | SNX1 can self-associate, but persistence through SDS preparation is unverified. | Compare denaturation conditions and confirm the band with SNX1 depletion. |
| Band lower than expected | An SNX1 splice isoform could migrate differently; its size is unspecified. | Confirm identity with SNX1 depletion and an isoform-specific control. |
| Multiple bands | Isoforms 1, 1A and 3 or phosphorylation could alter migration; distinct bands are unproven. | Compare signals after SNX1 depletion and use isoform-specific controls. |
| Weak or no signal | Recovery of peripheral membrane SNX1 may be low. | Check loading and extraction, then test a membrane-enriched fraction. |
Experiment evidence
Western blot images showing SNX1 band
M02692-1 · SNX1
A02692-2 · SNX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SNX1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SNX1 in Western blot, you can use cervix tissue, which has high HPA expression.
Positive control: Cervix (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cervix and ovary provide tissue controls, though membrane enrichment may improve SNX1 detection.
HPA tissue expression evidence for SNX1
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 |
|---|---|---|---|---|
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Placenta | decidual cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SNX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SNX1, answered from its protein features.
How should SNX1 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 SNX1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 1A and 3. Isoform 1A lacks canonical residues 91–155; isoform 3 replaces residues 507–522 with a longer sequence. These differences could affect band position, but the supplied features do not establish where either isoform migrates.
Decide whether to quantify total SNX1 or a particular isoform. An antibody targeting residues 91–155 may miss isoform 1A; one targeting the canonical 507–522 segment may miss isoform 3. Confirm the antibody's epitope before combining bands in one measurement.
Decide whether to quantify total SNX1 or a particular isoform. An antibody targeting residues 91–155 may miss isoform 1A; one targeting the canonical 507–522 segment may miss isoform 3. Confirm the antibody's epitope before combining bands in one measurement.
Which phosphorylation sites should I consider when interpreting SNX1 bands?
PTM · UniProt lists phosphoserine at 32, 39, 58, 72, 188 and 280, and phosphothreonine at 41 and 48. These are UniProt coordinates; paper or antibody numbering may differ. A site's presence does not prove that a particular band is phosphorylated or shifted.
Yes. UniProt lists N6-acetyllysine at position 237, using UniProt numbering. Consider this site when interpreting a modification-sensitive result, but do not assign an unexpected band to acetylation from position alone.
Yes. UniProt lists N6-acetyllysine at position 237, using UniProt numbering. Consider this site when interpreting a modification-sensitive result, but do not assign an unexpected band to acetylation from position alone.
Does this guide establish induction of SNX1?
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.
How should transfer be checked for SNX1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02692-1 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 SNX1 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Should SNX1 migrate exactly at its predicted mass?
Interpretation · The canonical sequence has a predicted mass of 59.1 kDa, but no observed band position is supplied. Use 59.1 kDa as a reference; the listed modifications alone do not establish a visible shift or explain any difference in apparent mass.
Compare candidate bands with the 59.1 kDa canonical prediction and the documented isoform sequence changes. Check the antibody epitope against regions 91–155 and 507–522. UniProt also lists phosphorylation and acetylation sites, but these features alone cannot identify an unexpected band.
SNX1 is listed as a peripheral membrane protein on the cytoplasmic side of endosome and trans-Golgi network membranes. Compare like fractions when quantifying it; a change in fraction abundance need not represent a change in total cellular SNX1.
Compare candidate bands with the 59.1 kDa canonical prediction and the documented isoform sequence changes. Check the antibody epitope against regions 91–155 and 507–522. UniProt also lists phosphorylation and acetylation sites, but these features alone cannot identify an unexpected band.
SNX1 is listed as a peripheral membrane protein on the cytoplasmic side of endosome and trans-Golgi network membranes. Compare like fractions when quantifying it; a change in fraction abundance need not represent a change in total cellular SNX1.
Boster reagents
SNX1 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-SNX1 Rabbit Monoclonal Antibody
Cat # M02692-1
Real WB data
Anti-SNX1 Antibody Picoband®
Cat # A02692-2
Two anti-SNX1 antibodies have WB images: M02692-1 in 293T lysate and A02692-2 in HeLa and HepG2 lysates. A02692-2 showed a band at 70–75 kDa, above the stated 59 kDa expected size; the images alone do not resolve that difference.
Which to pick: For human HeLa or HepG2 samples, A02692-2 has a documented WB example with conditions. M02692-1 lists human, mouse, and rat reactivity, but its supplied WB example uses only 293T lysate; verify performance in your sample.
Source: BosterBio SNX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13596.
- Human Protein Atlas. SNX1 tissue expression.
- PMC12758353 — target-verified WB comparison