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- Table of Contents
TYK2 / Non-receptor tyrosine-protein kinase TYK2 · Western blot design guide
Design a Western Blot for TYK2
Real validated TYK2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TYK2 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
TYK2 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 | ~133.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated TYK2 Western Blot Protocols
The A01642-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human small intestine tissue lysate (catalog A01642-1) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01642-1 · (A) 1 and (B) 2 μg/mL (catalog A01642-1) |
| 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 TYK2 Western Blot Band Size?
Full-length TYK2 is predicted at 133.7 kDa; phosphorylation may affect migration, but no empirical band or visible shift is established.
| Single band near 133.7 kDa | Consistent with predicted full-length TYK2; confirm its identity |
| Close doublet near 133.7 kDa | Could reflect TYK2 phosphorylation states, though a visible shift is unestablished |
| Several nearby bands | Could include different TYK2 phosphorylation states; confirm each band's identity |
| No band near 133.7 kDa | TYK2 was not detected at its predicted position; absence is unproven |
💡Expected TYK2 appearanceFull-length TYK2 has a predicted mass of 133.7 kDa, but no empirical band position is supplied; confirm any candidate band with ordinary band-identity controls.
| UniProt predicted mass | Places full-length TYK2 near 133.7 kDa; apparent migration is unestablished |
| Phosphotyrosine at residue 292 | May affect migration, but no visible shift is demonstrated |
| Phosphoserine at residue 499 | May affect migration, but no visible shift is demonstrated |
| Phosphotyrosine at residue 1054 | May affect migration, but no visible shift is demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TYK2 may be below detection or antibody binding may be inadequate | Check loading and antibody performance with a validated positive control |
| Band higher than expected | Band identity and reason for slower migration are unestablished | Compare with a validated TYK2 control and confirm with a second antibody |
| Band lower than expected | Proteolysis or an unrelated band is possible | Use fresh lysate with protease inhibitors and confirm with a second antibody |
| Multiple bands | TYK2 phosphorylation states are possible, but other bands may be unrelated | Compare phosphatase-treated and untreated lysates with appropriate controls |
| Weak or no signal | TYK2 signal may be below detection | Check sample loading and use a validated positive control |
Experiment evidence
Western blot images showing TYK2 band
A01642-1 · TYK2
A01642-3 · TYK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TYK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TYK2 in Western blot, you can use adrenal gland tissue, which shows high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports TYK2 as high in adrenal gland and not detected in caudate, making tissue controls feasible.
HPA tissue expression evidence for TYK2
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 → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced TYK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for TYK2, answered from its protein features.
Where should the TYK2 band appear?
Band shift · The supplied sequence predicts 133.7 kDa. Use that as a reference for locating the full-length band; no observed band position is provided. The listed modifications alone do not establish an apparent mass shift.
Could TYK2 isoforms explain multiple bands?
Isoforms · The supplied entry lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to TYK2 isoforms.
Which TYK2 phosphorylation sites are listed?
PTM · In UniProt coordinates, phosphotyrosines are listed at 292, 604, 1054, and 1055; phosphoserines at 499, 525, and 884. Check which coordinate convention a site-specific antibody uses before comparing its label with these positions.
UniProt lists phosphotyrosine 1054 as formed by autocatalysis. A phospho-specific measurement at this site addresses a modification, so measure total TYK2 separately when comparing protein abundance.
TYK2 has seven listed modified residues, but their presence alone does not demonstrate a visible shift or explain a measured mass difference. Compare matched samples with and without phosphatase treatment to test whether phosphorylation contributes to an observed shift.
UniProt lists phosphotyrosine 1054 as formed by autocatalysis. A phospho-specific measurement at this site addresses a modification, so measure total TYK2 separately when comparing protein abundance.
TYK2 has seven listed modified residues, but their presence alone does not demonstrate a visible shift or explain a measured mass difference. Compare matched samples with and without phosphatase treatment to test whether phosphorylation contributes to an observed shift.
Does this guide establish induction of TYK2?
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 TYK2 Western blot?
Transfer · TYK2 is predicted to be 133.7 kDa. Choose transfer conditions suitable for a protein of that size, and check the membrane and post-transfer gel to confirm that the TYK2 region transferred efficiently.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01642-1 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 TYK2 Western blot signals be quantified?
Quantitation · Quantify the full-length TYK2 band consistently across samples. If measuring a listed phosphorylation site, report its signal relative to total TYK2 as well; a phospho-specific signal alone does not measure total protein abundance.
How should unexpected TYK2 bands be interpreted?
Interpretation · Compare them with the predicted 133.7 kDa full-length protein and verify antibody specificity. The supplied entry lists one isoform and no alternative sequence; its phosphorylation features alone cannot identify an unexpected band or establish why it migrates differently.
Boster reagents
TYK2 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-TYK2 Antibody
Cat # A01642-1
Real WB data
Anti-TYK2 Antibody Picoband®
Cat # A01642-3
The catalog reports two anti-TYK2 antibodies with WB images: A01642-1 in human small intestine lysate at 1–2 μg/mL, and A01642-3 in human MCF-7 and HeLa lysates at 0.5 μg/mL, with a reported 134 kDa band. The images show human samples only.
Which to pick: Choose A01642-1 if catalogued mouse or rat reactivity matters, though its WB image uses human tissue. For human cell lysates, A01642-3 has images using MCF-7 and HeLa. Neither caption demonstrates mouse or rat WB.
Source: BosterBio TYK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P29597.
- Human Protein Atlas. TYK2 tissue expression.
- PMC11734966 — target-verified WB comparison
- PMC4288000 — target-verified WB comparison
- PMC8766497 — target-verified WB comparison