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- Table of Contents
SIN3A / Paired amphipathic helix protein Sin3a · Western blot design guide
Design a Western Blot for SIN3A
Real validated SIN3A Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SIN3A 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
SIN3A 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 | ~145.2 kDa | |
| Observed band | ~150 kDa | |
| Gel | 5–20% (catalog A01203-2) | |
| Positive control | Nasopharynx (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 | 1 isoform(s) |
Section 1
Real Curated SIN3A Western Blot Protocols
The A01203-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human Jurkat, human 293T (catalog A01203-2) |
| Gel % | 5–20% (catalog A01203-2) |
| Load | 30 ug (catalog A01203-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01203-2) |
| Membrane | nitrocellulose membrane (catalog A01203-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01203-2) |
| Primary antibody | A01203-2 · 0.5 μg/mL (catalog A01203-2) |
| Primary incubation | overnight at 4°C (catalog A01203-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01203-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01203-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01203-2) |
| Detection | ECL (catalog A01203-2) |
Section 2
What Is the Expected SIN3A Western Blot Band Size?
SIN3A is predicted at 145.2 kDa and observed near 150 kDa; the cause of the difference is not established.
| Band near 150 kDa | matches the reported SIN3A band; confirm identity with a specific control |
| Band near 145.2 kDa | is near the predicted full-length mass; confirm identity with a specific control |
| Faint band in whole-cell lysate | may reflect limited recovery of nuclear SIN3A |
| Close doublet near 150 kDa | could reflect modified SIN3A, although distinct migration is not established |
💡Expected SIN3A appearanceSIN3A has a predicted mass of 145.2 kDa and an empirical band near 150 kDa; the cause of the difference is unestablished, so confirm band identity with a specific control.
| Predicted full-length mass | 145.2 kDa by sequence; the reported band is near 150 kDa |
| Ser10 phosphorylation | documented site; an apparent size change is not established |
| Thr284 phosphorylation | documented site; an apparent size change is not established |
| Lys469 acetylation | documented site; an apparent size change is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear SIN3A may be poorly recovered | Check nuclear extraction and sample loading |
| Band higher than expected | The reported band is near 150 kDa versus 145.2 kDa predicted; the cause is unknown | Compare with the reported band and verify identity by SIN3A knockdown |
| Band lower than expected | Possible protein breakdown or nonspecific binding | Use fresh lysate with protease inhibitors and verify identity by SIN3A knockdown |
| Multiple bands | Modified SIN3A or nonspecific binding; distinct migration is unproven | Compare bands after SIN3A knockdown |
| Weak or no signal | Limited recovery of nuclear SIN3A | Check a nuclear fraction and confirm transfer of high-mass proteins |
| Fragments below expected size | Possible protein breakdown | Prepare fresh lysate with protease inhibitors and test fragment identity by SIN3A knockdown |
Experiment evidence
Western blot images showing SIN3A band
A01203-2 · SIN3A
M01203 · SIN3A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SIN3A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SIN3A in Western blot, you can use nasopharynx tissue, which HPA rates highly positive.
Positive control: Nasopharynx (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: SIN3A is nuclear, so nuclear-enriched lysate may improve detection; HPA lists adipose tissue as not detected.
HPA tissue expression evidence for SIN3A
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 |
|---|---|---|---|---|
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | cytotrophoblasts | High | Protein (IHC) | HPA → |
| Testis | preleptotene spermatocytes | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SIN3A Western Blot Tips
Deeper troubleshooting and optimisation questions for SIN3A, answered from its protein features.
How should SIN3A 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.
Are multiple SIN3A isoform bands expected?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Do not assign additional bands to specific SIN3A isoforms from this record alone.
Which phosphorylation sites matter when interpreting SIN3A bands?
PTM · UniProt lists phosphoserine at 10, 277, 832, 860, 940, 1089, and 1112, plus phosphothreonine at 284. These are UniProt coordinates; antibody or paper numbering may differ. The listed sites do not establish a visible band shift.
UniProt lists N6-acetyllysine at positions 469, 865, and 875. These are UniProt coordinates. Their presence alone does not show that acetylation caused a separate band; compare the band with the expected approximately 150 kDa signal.
UniProt lists N6-acetyllysine at positions 469, 865, and 875. These are UniProt coordinates. Their presence alone does not show that acetylation caused a separate band; compare the band with the expected approximately 150 kDa signal.
Does this guide establish induction of SIN3A?
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 SIN3A Western blot?
Transfer · SIN3A is a large protein: its calculated mass is 145.2 kDa, with an observed band near 150 kDa. Choose and verify transfer conditions that retain a clear signal at approximately 150 kDa.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01203-2 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 nuclear localization guide SIN3A quantitation?
Quantitation · UniProt places SIN3A in the nucleus and nucleolus and notes recruitment to the nucleolus by SAP30L. Compare like fractions across samples when quantifying its band; changes in fraction composition can affect the measured signal.
Why might SIN3A appear near 150 kDa instead of 145.2 kDa?
Interpretation · The calculated mass is 145.2 kDa, and the supplied observed band is approximately 150 kDa. SIN3A has listed phosphorylation and acetylation sites, but their presence alone does not establish the cause of this difference.
Start with the approximately 150 kDa observed band. The record lists one isoform and no signal peptide or propeptide. Its listed modifications and protein interactions do not, by themselves, identify the source of another band.
Start with the approximately 150 kDa observed band. The record lists one isoform and no signal peptide or propeptide. Its listed modifications and protein interactions do not, by themselves, identify the source of another band.
Boster reagents
SIN3A 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-mSin3A/SIN3A Antibody Picoband®
Cat # A01203-2
Real WB data
Anti-mSin3A Rabbit Monoclonal Antibody
Cat # M01203
Two the supplier anti-SIN3A antibodies have Western blot images. A01203-2 shows a band near 150 kDa in five human cell lysates and rat PC-12 lysate; M01203 shows blots of K562 and RAW 264.7 lysates. The supplied evidence is limited to these product examples.
Which to pick: For the documented human and rat samples, consider A01203-2. For mouse work, M01203 lists mouse reactivity and shows a RAW 264.7 blot. Both have WB images; choose based on your sample and the reported conditions.
Source: BosterBio SIN3A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96ST3.
- Human Protein Atlas. SIN3A tissue expression.
- PMC12889623 — target-verified WB comparison
- PMC7661264 — target-verified WB comparison
- PMC10157306 — target-verified WB comparison