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- Table of Contents
SMS / Spermine synthase · Western blot design guide
Design a Western Blot for SMS
Real validated SMS Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SMS 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
SMS 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 | ~41.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Ovary (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SMS Western Blot Protocols
The M01831-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M01831-1) |
| 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 | M01831-1; 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 SMS Western Blot Band Size?
SMS has a predicted 41.3 kDa monomer; homodimerization, splicing, and phosphorylation could affect patterns, but altered migration is unproven and no empirical band size is supplied.
| Single band near 41.3 kDa | consistent with the predicted SMS monomer; confirm identity with antibody controls |
| Band near 83 kDa | could represent a retained SMS homodimer if sample preparation preserves it |
| Several distinct bands | could reflect isoforms 1 and 2 if they migrate differently; their separation is unestablished |
| Close doublet near 41.3 kDa | could reflect Ser57 phosphorylation, but the modification does not establish a visible shift |
💡Expected SMS appearanceThe predicted SMS monomer is 41.3 kDa; no empirical band size is supplied, so confirm any candidate band with antibody identity controls.
| UniProt predicted monomer mass | sets a 41.3 kDa reference, not a measured migration |
| SMS homodimer | could appear near twice the monomer mass if the complex survives sample preparation |
| Splice isoform 1 | its size relative to isoform 2 is not supplied |
| Splice isoform 2 | its size relative to isoform 1 is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | an SMS homodimer may have survived sample preparation | compare denaturing conditions and confirm the band with antibody controls |
| Band lower than expected | isoform size differences are possible but unestablished | verify the band with an independent SMS antibody or isoform-specific evidence |
| Multiple bands | isoforms 1 and 2 may migrate differently, although separation is unestablished | check band identity with an independent antibody or SMS depletion |
| Weak or no signal | SMS abundance or antibody detection in this sample may be insufficient | check loading, antibody performance, and an SMS-positive control |
| Broad smear instead of sharp band | the supplied SMS features do not establish a biological cause | check sample integrity and antibody background with appropriate controls |
Experiment evidence
Western blot images showing SMS band
M01831-1 · SMS
A01831-1 · SMS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SMS Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SMS in Western blot, you can use ovary lysate, which HPA scores high for SMS expression.
Positive control: Ovary (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: HPA lists high-expression and not-detected tissues, making tissue controls feasible.
HPA tissue expression evidence for SMS
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 |
|---|---|---|---|---|
| Ovary | follicle cells | High | Protein (IHC) | HPA → |
| Testis | peritubular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Placenta | trophoblastic 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SMS Western Blot Tips
Deeper troubleshooting and optimisation questions for SMS, answered from its protein features.
What band size should I expect for SMS?
Band shift · The canonical SMS sequence is 366 residues with a predicted mass of 41.3 kDa. No observed Western blot band size is supplied, so use 41.3 kDa as a reference rather than an established apparent mass.
Could SMS isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 57–109 of the canonical sequence, so it may migrate differently. The supplied features do not establish its observed band position.
No. Isoform 2 lacks canonical residues 57–109, which include the listed phosphoserine at position 57. Keep the UniProt coordinate convention explicit when comparing sites reported with other numbering.
No. Isoform 2 lacks canonical residues 57–109, which include the listed phosphoserine at position 57. Keep the UniProt coordinate convention explicit when comparing sites reported with other numbering.
Which SMS modifications matter when interpreting bands?
PTM · The listed modifications are N-acetylalanine at position 2 and phosphoserine at position 57, using UniProt canonical coordinates. Their presence alone does not establish a visible band shift or explain any difference from the predicted mass.
Does this guide establish induction of SMS?
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 SMS?
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 M01831-1 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 SMS 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.
How should I compare SMS abundance across samples?
Interpretation · Decide whether the measurement should include both listed isoforms. Because isoform 2 lacks canonical residues 57–109, an antibody recognizing that region could miss it. Check the antibody epitope and quantify isoform bands separately if both are detected.
Consider isoform 2, which lacks canonical residues 57–109. The supplied features do not provide its observed migration or establish that any particular lower band is SMS; confirm the band's identity before assigning it to an isoform.
SMS is described as a homodimer mediated by its N-terminal domain. That feature does not show that a dimer remains intact during Western blot preparation or predict a higher band. Confirm band identity before assigning a higher band to a dimer.
Consider isoform 2, which lacks canonical residues 57–109. The supplied features do not provide its observed migration or establish that any particular lower band is SMS; confirm the band's identity before assigning it to an isoform.
SMS is described as a homodimer mediated by its N-terminal domain. That feature does not show that a dimer remains intact during Western blot preparation or predict a higher band. Confirm band identity before assigning a higher band to a dimer.
Boster reagents
SMS 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-Spermine synthase Rabbit Monoclonal Antibody
Cat # M01831-1
Real WB data
Anti-Spermine synthase/SMS Antibody Picoband®
Cat # A01831-1
Two the supplier anti-SMS antibodies have Western blot images: M01831-1 in HeLa lysate and A01831-1 in human U-87MG, U251, and K562 lysates, with a reported 41 kDa band. The supplied evidence does not establish performance across all listed species.
Which to pick: For human samples, both have WB images; choose A01831-1 if its documented 41 kDa band and detailed conditions are useful. M01831-1 also lists Mouse and Rat reactivity, but its supplied WB image uses HeLa lysate only.
Source: BosterBio SMS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P52788.
- Human Protein Atlas. SMS tissue expression.
- PMC4428506 — target-verified WB comparison
- PMC11577379 — target-verified WB comparison
- PMC7320137 — target-verified WB comparison