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- Table of Contents
TIA1 / Cytotoxic granule associated RNA binding protein TIA1 · Western blot design guide
Design a Western Blot for TIA1
Real validated TIA1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TIA1 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
TIA1 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 | ~43 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Isoform-aware interpretation | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated TIA1 Western Blot Protocols
The PA2194 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | JURKAT, RAJI, CEM (catalog PA2194) |
| 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 | PA2194; 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 TIA1 Western Blot Band Size?
TIA1 is predicted at 43 kDa; isoforms could affect band patterns, but distinct migration has not been demonstrated by the supplied evidence.
| Band near 43 kDa | Consistent with the predicted size of TIA1; confirm identity with antibody controls. |
| Additional band below the main band | Could reflect the Short isoform; its migration is not established. |
| Several discrete bands | Could reflect Long, Short, and 3 isoforms if their migration differs. |
| Higher band under incomplete denaturation | Could reflect Zn(2+)-induced TIA1 homooligomerization; its gel migration is unverified. |
💡Expected TIA1 appearanceUniProt predicts TIA1 at 43 kDa, but no empirical band size is supplied; identify a candidate band with a validated positive control and an independent TIA1 antibody.
| UniProt predicted mass | Full-length TIA1 has a predicted mass of 43 kDa; apparent migration is unverified. |
| Long isoform | May differ in size from other isoforms; no isoform mass is supplied. |
| Short isoform | May migrate differently from Long; no isoform mass is supplied. |
| Isoform 3 | May migrate differently from the other isoforms; its relative size is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TIA1 may be poorly recovered from nuclear or stress granule pools. | Check nuclear and cytoplasmic fractions alongside a validated positive lysate. |
| Band higher than expected | Zn(2+)-induced homooligomerization is possible; its effect on this blot is unverified. | Check denaturation conditions and confirm the band with an independent TIA1 antibody. |
| Band lower than expected | A Short isoform is possible, but its band position is unknown. | Compare isoform-aware controls or confirm the band with a second antibody. |
| Multiple bands | Long, Short, and 3 isoforms may differ in migration. | Use isoform-aware controls and a second antibody to assign bands. |
| Weak or no signal | Recovery may vary between nuclear, cytoplasmic, and stress granule pools. | Check fraction recovery and compare with a validated positive lysate. |
Experiment evidence
Western blot images showing TIA1 band
PA2194 · TIA1
M02763-1 · TIA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TIA1 Western blot
🧪For positive controls for TIA1 in Western blot, you can use a TIA1-positive lysate verified independently, since no HPA positive tissue or cell is supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA supplies no tissue expression data, so tissue controls cannot be selected from this evidence.
HPA tissue expression evidence for TIA1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced TIA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TIA1, answered from its protein features.
How should TIA1 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.
Which TIA1 isoforms could produce different bands?
Isoforms · Three isoforms are listed: Long, Short, and 3. Short replaces canonical residues 93–104 with N. Isoform 3 replaces residues 195–214 and lacks residues 215–386. These sequence differences could change band size and antibody recognition; the supplied data do not establish their observed migration.
Does TIA1 have a documented modification site?
PTM · Yes. UniProt lists N-acetylmethionine at position 1. This is a UniProt sequence coordinate; antibody or paper numbering may differ. The feature alone does not demonstrate a visible band shift.
The supplied interaction annotation says TIA1 interacts with FASTK and that the interaction leads to TIA1 phosphorylation. No phosphorylation coordinate is given here. Compare relevant conditions if investigating phosphorylation, but do not assign a band shift to it without supporting evidence.
The supplied interaction annotation says TIA1 interacts with FASTK and that the interaction leads to TIA1 phosphorylation. No phosphorylation coordinate is given here. Compare relevant conditions if investigating phosphorylation, but do not assign a band shift to it without supporting evidence.
Does this guide establish induction of TIA1?
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 TIA1?
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 PA2194 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 TIA1 bands be quantified across samples?
Quantitation · Use the same sample fraction and band selection across samples. Because three isoforms are listed, specify whether quantitation includes one band or multiple bands, and check whether the antibody can recognize the relevant sequences. The supplied features do not identify an antibody epitope.
Should TIA1 migrate at exactly 43 kDa?
Interpretation · The supplied predicted mass is 43 kDa, but no observed band position is available. Use 43 kDa as a starting reference, not an exact migration requirement. The listed features alone do not establish a visible shift.
TIA1 is annotated in the nucleus, cytoplasm, and cytoplasmic stress granules. State whether the measurement uses whole-cell lysate or a fraction, and compare like fractions across samples; redistribution could change a fraction's signal without establishing a change in total TIA1.
Consider the documented isoforms first: Short changes residues 93–104, while isoform 3 replaces residues 195–214 and lacks 215–386. TIA1 also has N-terminal acetylation at UniProt position 1 and a FASTK-linked phosphorylation annotation. None of these features alone identifies an unexpected band; confirm its identity experimentally.
TIA1 is annotated in the nucleus, cytoplasm, and cytoplasmic stress granules. State whether the measurement uses whole-cell lysate or a fraction, and compare like fractions across samples; redistribution could change a fraction's signal without establishing a change in total TIA1.
Consider the documented isoforms first: Short changes residues 93–104, while isoform 3 replaces residues 195–214 and lacks 215–386. TIA1 also has N-terminal acetylation at UniProt position 1 and a FASTK-linked phosphorylation annotation. None of these features alone identifies an unexpected band; confirm its identity experimentally.
Boster reagents
TIA1 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-Nucleolysin TIA-1 isoform p40 TIA1 Antibody Picoband®
Cat # PA2194
Real WB data
Anti-TIA1 Rabbit Monoclonal Antibody
Cat # M02763-1
Both catalogued anti-TIA1 antibodies, PA2194 and M02763-1, list Human and Mouse reactivity and have Western blot images. The supplied evidence consists of product captions; no independent publication evidence is provided.
Which to pick: For mouse lysates, M02763-1 shows an NIH/3T3 blot. For human lysates, both show Jurkat; PA2194 also shows RAJI, CEM, HT1080, and K562. Pick the antibody with the closest shown sample context; these images do not establish performance in every sample.
Source: BosterBio TIA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P31483.
- Human Protein Atlas. TIA1 tissue expression.
- PMC6377165 — target-verified WB comparison
- PMC8437838 — target-verified WB comparison
- PMC5336638 — target-verified WB comparison