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- Table of Contents
SON / Protein SON · Western blot design guide
Design a Western Blot for SON
Real validated SON Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SON 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
SON 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 | ~263.8 kDa | |
| Observed band | ~264 kDa | |
| Gel | 8% (catalog A00288-1) | |
| Positive control | Adrenal gland (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 | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 10 isoform(s) |
Section 1
Real Curated SON Western Blot Protocols
The A00288-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human K562, human SIHA (catalog A00288-1) |
| Gel % | 8% (catalog A00288-1) |
| Load | 30 ug; reducing conditions (catalog A00288-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00288-1) |
| Membrane | nitrocellulose membrane (catalog A00288-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00288-1) |
| Primary antibody | A00288-1 · 0.5 μg/mL (catalog A00288-1) |
| Primary incubation | overnight at 4°C (catalog A00288-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00288-1) |
| Secondary incubation | 1.5 hour at RT (catalog A00288-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00288-1) |
| Detection | ECL (catalog A00288-1) |
Section 2
What Is the Expected SON Western Blot Band Size?
SON is predicted at 263.8 kDa and observed at ~264 kDa; isoforms and modifications are annotated, but their effects on migration are not established.
| Band at ~264 kDa | Consistent with SON in whole-cell lysate; confirm identity with appropriate controls |
| Band at ~264 kDa in a nuclear fraction | Consistent with SON localization to nuclear speckles |
| Additional bands at different positions | Could reflect SON isoforms; their migration is not established |
| Little or no band in a cytoplasmic fraction | Consistent with SON's nuclear localization |
💡Expected SON appearanceSON has a predicted mass of 263.8 kDa and an empirical band at ~264 kDa in reducing whole-cell blots; confirm band identity with appropriate controls.
| Predicted SON mass | 263.8 kDa, consistent with the observed ~264 kDa band |
| Isoforms F, A, B, C, D, E, G, H, I and J | May differ in size, but their individual masses and resolved bands are not established |
| Phosphorylation at Ser94, Ser142, Ser152, Ser154, Ser160 and Ser283 | May affect mobility; a visible shift is not established |
| Acetylation at Ala2 and Lys16 | May affect mobility; a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear SON may be poorly extracted | Check nuclear extraction and probe a nuclear fraction |
| Band higher than expected | An alternative isoform or modification is possible, but its effect is unproven | Compare with the ~264 kDa reference and confirm identity by SON depletion |
| Band lower than expected | An alternative isoform or proteolysis is possible | Use protease inhibitors and confirm identity by SON depletion |
| Multiple bands | SON has ten named isoforms, though distinct bands are unverified | Compare isoform-specific expression or use SON depletion to identify related bands |
| Weak or no signal | Nuclear extraction or transfer of this large protein may be inadequate | Check nuclear recovery and transfer near 264 kDa |
| Fragments below expected size | Proteolysis of the large SON protein is possible | Process samples promptly with protease inhibitors and compare fresh lysate |
Experiment evidence
Western blot images showing SON band
A00288-1 · SON
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SON Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SON in Western blot, you can use adrenal gland tissue, which shows high HPA expression.
Positive control: Adrenal gland (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.
⚠️Feasibility: SON is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for SON
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 → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | 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 → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SON Western Blot Tips
Deeper troubleshooting and optimisation questions for SON, answered from its protein features.
How should SON 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.
Which SON isoforms could produce smaller bands?
Isoforms · UniProt lists 10 isoforms. Relative to its 2426-residue reference sequence, isoform H lacks residues 690–2416; A lacks 688–1006; E lacks 2109–2426. B, C, and J also have C-terminal deletions. These sequence differences can change expected mass, but they do not establish which isoform is present in a sample.
Map the antibody's epitope to the UniProt reference sequence. An epitope within residues 688–1006 is absent from isoform A, while one within 690–2416 is absent from isoform H. Other listed variants alter C-terminal regions. An antibody may therefore detect only some SON isoforms.
Map the antibody's epitope to the UniProt reference sequence. An epitope within residues 688–1006 is absent from isoform A, while one within 690–2416 is absent from isoform H. Other listed variants alter C-terminal regions. An antibody may therefore detect only some SON isoforms.
Which SON modifications matter when interpreting bands?
PTM · UniProt lists 42 modified residues, including phosphoserines at reference-sequence positions 94, 1948, and 2238; asymmetric dimethylarginines at 1007 and 1022; and N6-acetyllysine at 16 and 288. Residue 2055 has an alternate N6-acetyllysine annotation. Use UniProt reference coordinates when comparing site-specific reagents; these annotations alone do not prove a band shift.
No. SON has many annotated phosphorylation sites, including reference-sequence serines 1948, 1950, 1952, 2009, 2011, and 2013. Their presence does not show that a particular sample is phosphorylated or that phosphorylation caused its apparent mobility. Confirm the modification with a site-specific assay before assigning the band.
No. SON has many annotated phosphorylation sites, including reference-sequence serines 1948, 1950, 1952, 2009, 2011, and 2013. Their presence does not show that a particular sample is phosphorylated or that phosphorylation caused its apparent mobility. Confirm the modification with a site-specific assay before assigning the band.
Does this guide establish induction of SON?
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 SON?
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 A00288-1 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.
What matters when quantifying SON across samples?
Quantitation · SON localizes to nuclear speckles, and its listed isoforms differ substantially in sequence. Compare equivalent sample preparations and define which antibody-reactive band or bands you measure. If several bands are present, report them separately unless their SON identity and isoform coverage have been established.
Does SON's observed band differ from its predicted mass?
Interpretation · The reported band at approximately 264 kDa closely matches the predicted 263.8 kDa. Compare additional bands with the expected size of the isoform your antibody recognizes; modification sites alone do not establish a visible shift.
First compare them with the approximately 264 kDa observed band and with the sequence changes of the 10 listed isoforms. Consider whether the antibody's epitope is retained in each candidate isoform. SON also has annotated modifications, but the supplied features cannot identify the cause of any additional band.
First compare them with the approximately 264 kDa observed band and with the sequence changes of the 10 listed isoforms. Consider whether the antibody's epitope is retained in each candidate isoform. SON also has annotated modifications, but the supplied features cannot identify the cause of any additional band.
Boster reagents
SON 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-SON Antibody Picoband®
Cat # A00288-1
The catalog reports A00288-1 for SON Western blotting, with stated human and mouse reactivity. Its WB image caption reports a band near the expected 264 kDa in four human cell lysates; no mouse WB example is supplied.
Which to pick: A00288-1 is the only listed option. It has a WB image using human HEL, K562, SIHA, and MCF-7 lysates at 0.5 μg/mL. Mouse reactivity is listed, but the supplied image documents human samples only.
Source: BosterBio SON gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P18583.
- Human Protein Atlas. SON tissue expression.
- PMC10680872 — target-verified WB comparison
- PMC10972584 — target-verified WB comparison
- PMC8155101 — target-verified WB comparison