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- Table of Contents
PTS / 6-pyruvoyl tetrahydrobiopterin synthase · Western blot design guide
Design a Western Blot for PTS
Real validated PTS Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTS 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
PTS 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 | ~16.4 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Endometrium (total-target IHC; phospho state unverified) +4 more | |
| Negative control | Adipose tissue (total-target IHC; phospho state unverified) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphatase-treated control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PTS Western Blot Protocols
The P00430 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (1) HUVEC cell lysate; (2) HUVEC cell treated with Alkaline Phosphatase lysate (catalog P00430) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% BSA in TBST (standard starting point) |
| Primary antibody | P00430; 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 PTS Western Blot Band Size?
PTS has a predicted 16.4 kDa monomer; phosphorylation and homohexamer formation are reported, but their effects on Western-blot migration are unproven.
| Band near 16.4 kDa | Consistent with PTS monomer; confirm identity with controls |
| Band that weakens after phosphatase treatment | Consistent with recognition of phosphorylated PTS at S19 |
| No band with the phospho-S19 antibody | PTS may lack detectable S19 phosphorylation in this sample |
| Higher band under conditions preserving protein complexes | Could reflect the reported PTS homohexamer; identity requires confirmation |
💡Expected PTS appearanceUniProt predicts a 16.4 kDa PTS monomer, but no empirical band size is supplied; the lead antibody recognizes phospho-S19 PTS, so confirm any candidate band with appropriate identity and phosphatase controls.
| Predicted PTS mass | Places the monomer near 16.4 kDa |
| Homohexamer assembly | May produce a higher apparent size if the complex remains intact |
| Phosphoserine 19 | Defines the lead antibody's target state; no migration shift is established |
| Phosphoserine 28 | Is reported, but no effect on migration is established |
| Phosphotyrosine 128 | Is reported, but no effect on migration is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | S19 phosphorylation may be undetectable | Compare with a total PTS antibody and a suitable positive control |
| Band higher than expected | PTS may retain its reported oligomeric assembly | Check denaturation conditions and confirm band identity |
| Band lower than expected | Band identity or protein integrity is uncertain | Compare with a total PTS antibody and inspect sample handling |
| Multiple bands | Their identities are not established by the supplied features | Use phosphatase treatment and independent PTS detection to identify the target band |
| Weak or no signal | The phospho-S19 epitope may be scarce or lost | Include a phospho-positive control and compare with total PTS |
| Fragments below expected size | Protein degradation is possible | Check sample handling and confirm fragments with an independent PTS antibody |
Experiment evidence
Western blot images showing PTS band
P00430 · PTS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PTS Western blot
🧪HPA-IHC candidate guidance (verify in WB): HPA tissue expression does not establish phosphorylation at the selected antibody epitope. Verify activation-state controls for the phospho-specific lead. HPA-IHC candidate guidance (verify in WB): For positive controls for PTS in Western blot, you can use endometrium tissue, which HPA scores as medium expression.
Positive control: Endometrium (total-target IHC; phospho state unverified)
Negative control: Adipose tissue (total-target IHC; phospho state unverified)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA identifies a not-detected tissue for a negative control, but the positive tissue has only medium expression.
HPA tissue expression evidence for PTS
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 |
|---|---|---|---|---|
| Endometrium | cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Medium | Protein (IHC) | HPA → |
| Kidney | cells in tubules | Medium | Protein (IHC) | HPA → |
| Liver | hepatocytes | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTS Western Blot Tips
Deeper troubleshooting and optimisation questions for PTS, answered from its protein features.
Where should the main PTS band appear?
Band shift · PTS has a predicted mass of 16.4 kDa. Use that as a starting point for identifying the band; no observed band position is supplied, so its apparent mass cannot be confirmed here.
Are distinct PTS isoform bands expected?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning a second band to a distinct PTS isoform.
Could phosphorylation shift the PTS band?
PTM · UniProt lists phosphoserine at positions 19 and 28 and phosphotyrosine at 128. These modifications may affect apparent migration, but their presence alone does not establish a visible shift or explain a difference from 16.4 kDa.
Using the supplied UniProt sequence numbering, Ser19 is phosphorylated by PKG, Ser28 is phosphorylated, and Tyr128 is phosphorylated. The supplied features do not name an enzyme for Ser28 or Tyr128; check numbering conventions before comparing antibody labels.
Measure a site-specific phospho signal alongside total PTS when assessing Ser19, Ser28, or Tyr128. Report the site using UniProt numbering. A total PTS band alone does not show which listed residue is phosphorylated.
Using the supplied UniProt sequence numbering, Ser19 is phosphorylated by PKG, Ser28 is phosphorylated, and Tyr128 is phosphorylated. The supplied features do not name an enzyme for Ser28 or Tyr128; check numbering conventions before comparing antibody labels.
Measure a site-specific phospho signal alongside total PTS when assessing Ser19, Ser28, or Tyr128. Report the site using UniProt numbering. A total PTS band alone does not show which listed residue is phosphorylated.
Does this guide establish induction of PTS?
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 PTS?
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 P00430 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 PTS 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 a higher molecular weight PTS band be interpreted?
Interpretation · PTS is described as a homohexamer made of two homotrimers. A higher band could prompt a check for retained assemblies, but these features do not establish that an assembly survives the blot conditions or identify any particular band.
Boster reagents
PTS 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-Phospho-PTS (S19) Rabbit Monoclonal Antibody
Cat # P00430
The catalog reports one anti-PTS antibody: P00430, a human-reactive phospho-PTS (S19) rabbit monoclonal. Its Western blot image compares HUVEC lysate with alkaline-phosphatase-treated HUVEC lysate. The supplied evidence is limited to that comparison.
Which to pick: P00430 is the only listed SKU. Choose it for a phospho-PTS (S19) Western blot in the reported human HUVEC context; the supplied image does not establish performance in other samples.
Source: BosterBio PTS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q03393.
- Human Protein Atlas. PTS tissue expression.
- PMC9554678 — target-verified WB comparison