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- Table of Contents
TYMS / Thymidylate synthase · Western blot design guide
Design a Western Blot for TYMS
Real validated TYMS Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TYMS 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
TYMS 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 | ~35.7 kDa | |
| Observed band | ~36 kDa | |
| Gel | 5–20% (catalog A04320-4) | |
| Positive control | Bone marrow (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated TYMS Western Blot Protocols
The A04320-4 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, monkey COS-7 (catalog A04320-4) |
| Gel % | 5–20% (catalog A04320-4) |
| Load | 30 ug; reducing conditions (catalog A04320-4) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04320-4) |
| Membrane | nitrocellulose membrane (catalog A04320-4) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04320-4) |
| Primary antibody | A04320-4 · 0.5 μg/mL (catalog A04320-4) |
| Primary incubation | overnight at 4°C (catalog A04320-4) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04320-4) |
| Secondary incubation | 1.5 hour at RT (catalog A04320-4) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04320-4) |
| Detection | ECL (catalog A04320-4) |
Section 2
What Is the Expected TYMS Western Blot Band Size?
TYMS is predicted at 35.7 kDa and observed at ~36 kDa; the cause of their small apparent difference is not established.
| Band at ~36 kDa | Empirical TYMS band, close to its 35.7 kDa predicted mass |
| Band near ~72 kDa | Possible retained TYMS homodimer if it survives electrophoresis |
| Several bands at different positions | Could reflect TYMS isoforms 1, 2, and 3; distinct migration is unconfirmed |
| Weak band in one cell fraction | TYMS occurs in the nucleus, cytoplasm, and mitochondrion |
💡Expected TYMS appearanceTYMS has a predicted mass of 35.7 kDa and an empirical band at ~36 kDa in reducing whole-cell lysates; confirm band identity with appropriate antibody and sample controls.
| Predicted TYMS mass | 35.7 kDa, close to the observed ~36 kDa band |
| TYMS homodimer | Could appear near twice the monomer mass if retained during electrophoresis |
| Isoform 1 | Mass relative to other isoforms is not supplied |
| Isoform 2 | Mass relative to other isoforms is not supplied |
| Isoform 3 | Mass relative to other isoforms is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A TYMS homodimer may have survived sample preparation | Compare reducing and nonreducing samples and confirm band identity |
| Band lower than expected | An isoform is possible, but its migration is unknown | Check antibody specificity and compare with the ~36 kDa control band |
| Multiple bands | TYMS has three annotated isoforms, but distinct bands are unconfirmed | Use isoform-specific controls to identify the bands |
| Weak or no signal | TYMS distribution across cell compartments may affect fraction signal | Check loading and compare whole-cell and compartment fractions |
| Fragments below expected size | Fragment identity is not established by the supplied features | Check sample integrity and confirm smaller bands with an independent antibody |
Experiment evidence
Western blot images showing TYMS band
A04320-4 · TYMS
A04320-3 · TYMS
M04320-1 · TYMS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TYMS Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TYMS in Western blot, you can use bone marrow lysate, which HPA rates High.
Positive control: Bone marrow (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 the samples.
⚠️Feasibility: TYMS is intracellular, so lysate controls are feasible, with HPA not-detected adipose tissue as the negative comparator.
HPA tissue expression evidence for TYMS
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
| Testis | preleptotene spermatocytes | High | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | 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 → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TYMS Western Blot Tips
Deeper troubleshooting and optimisation questions for TYMS, answered from its protein features.
How should TYMS 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 TYMS isoforms produce additional bands?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 lacks residues 152–185 and isoform 3 lacks residues 69–151. These deletions could affect apparent size, but the features alone do not establish that separate bands will be visible.
What modification should I consider when interpreting TYMS bands?
PTM · UniProt lists phosphoserine at canonical residue 114. Its presence does not establish a visible band shift. Canonical residue 114 falls within the segment missing from isoform 3, so keep the isoform and coordinate convention explicit when assessing a phospho-specific signal.
No glycosylation sites are listed for TYMS in the supplied features. Do not attribute an unexpected band to glycosylation on this evidence alone.
No glycosylation sites are listed for TYMS in the supplied features. Do not attribute an unexpected band to glycosylation on this evidence alone.
Does this guide establish induction of TYMS?
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 TYMS?
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 A04320-4 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 TYMS 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 does the observed TYMS band compare with predicted mass?
Interpretation · The observed band is about 36 kDa, close to the 35.7 kDa predicted mass. This agreement does not establish which isoform or modification state produced it.
TYMS is annotated as a homodimer and as part of a complex with SHMT1 and DHFR. These associations alone do not identify a higher Western blot band; verify its TYMS identity before assigning it to a dimer or complex.
TYMS is listed in the nucleus, cytoplasm, and mitochondrion, including the matrix and inner membrane. When comparing fractions, identify the fraction used and quantify TYMS consistently within it; a change in fraction abundance need not represent the same change across the whole cell.
Check whether it could correspond to an annotated isoform: isoform 2 lacks canonical residues 152–185, while isoform 3 lacks 69–151. A smaller band alone cannot establish isoform identity.
TYMS is annotated as a homodimer and as part of a complex with SHMT1 and DHFR. These associations alone do not identify a higher Western blot band; verify its TYMS identity before assigning it to a dimer or complex.
TYMS is listed in the nucleus, cytoplasm, and mitochondrion, including the matrix and inner membrane. When comparing fractions, identify the fraction used and quantify TYMS consistently within it; a change in fraction abundance need not represent the same change across the whole cell.
Check whether it could correspond to an annotated isoform: isoform 2 lacks canonical residues 152–185, while isoform 3 lacks 69–151. A smaller band alone cannot establish isoform identity.
Boster reagents
TYMS 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-Thymidylate Synthase/TYMS Antibody Picoband®
Cat # A04320-4
Real WB data
Anti-TYSY Antibody(C-term)
Cat # A04320-3
Real WB data
Anti-Thymidylate Synthase TYMS Monoclonal Antibody
Cat # M04320-1
Three the supplier anti-TYMS antibodies have Western blot images. A04320-4 and M04320-1 show a band near the expected 36 kDa; A04320-3 shows detection in human cell lysates without a reported band size. These images document the listed samples and conditions, not broader validation.
Which to pick: For human samples, compare the three images and reported conditions. Choose M04320-1 if you need an image including human, monkey, rat, and mouse samples. A04320-4 shows human and monkey lysates; A04320-3 shows human cell lysates only.
Source: BosterBio TYMS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P04818.
- Human Protein Atlas. TYMS tissue expression.
- PMC6290025 — target-verified WB comparison
- PMC4695198 — target-verified WB comparison
- PMC12682279 — target-verified WB comparison