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- Table of Contents
TNNT1 / Troponin T, slow skeletal muscle · Western blot design guide
Design a Western Blot for TNNT1
Source-linked TNNT1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TNNT1 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
TNNT1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~32.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Skeletal muscle (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (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 | 3 isoform(s) |
Section 1
Source-Linked TNNT1 Western Blot Protocol Options
The A05799T1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A549, sp2/0, Rat skeletal muscel (catalog A05799T1) |
| 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 | A05799T1; 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 TNNT1 Western Blot Band Size?
TNNT1 is predicted at 32.9 kDa; three isoforms and Ser2 phosphorylation are annotated, but their effects on band migration are unproven.
| Band near 32.9 kDa | Consistent with the predicted TNNT1 mass; confirm identity with controls. |
| Single sharp band | Compatible with TNNT1; the three annotated isoforms need not resolve separately. |
| Several bands at different positions | Isoforms 1, 2, and 3 are possible contributors; band identities require validation. |
| Close doublet | Ser2 phosphorylation by CK2 is a possible contributor, but a mobility effect is unproven. |
💡Expected TNNT1 appearanceTNNT1 has a predicted mass of 32.9 kDa, but no empirical band size is supplied; use an appropriate positive control and identity checks to assess any detected band.
| Predicted TNNT1 mass | The reference sequence has a calculated mass of 32.9 kDa. |
| Splice isoform 1 | Its apparent size relative to the other isoforms is unknown. |
| Splice isoform 2 | Its apparent size relative to the other isoforms is unknown. |
| Splice isoform 3 | Its apparent size relative to the other isoforms is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TNNT1 may be scarce in the sampled material. | Run skeletal muscle lysate as a positive control. |
| Band higher than expected | An isoform may differ in size, but its migration is unknown. | Verify band identity with a second antibody or TNNT1 depletion. |
| Band lower than expected | An isoform or sample degradation may contribute; neither is established. | Compare fresh samples and verify band identity. |
| Multiple bands | Three splice isoforms are annotated, but their band positions are unknown. | Check which bands respond to TNNT1 depletion. |
| Weak or no signal | Low TNNT1 abundance or assay sensitivity may limit detection. | Check a skeletal muscle positive control and antibody conditions. |
| Fragments below expected size | Sample proteolysis is possible, though no fragment is specified. | Prepare fresh lysate with protease inhibitors and compare band patterns. |
Experiment evidence
Western blot images showing TNNT1 band
A05799T1 · TNNT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TNNT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TNNT1 in Western blot, you can use skeletal muscle lysate, which HPA rates high for TNNT1.
Positive control: Skeletal muscle (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: Tissue controls are feasible because HPA rates skeletal muscle high and adipose tissue not detected.
HPA tissue expression evidence for TNNT1
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 |
|---|---|---|---|---|
| Skeletal muscle | myocytes | High | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TNNT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TNNT1, answered from its protein features.
How should TNNT1 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 TNNT1 isoforms could contribute to multiple bands?
Isoforms · UniProt lists isoforms 1, 2 and 3. Relative to the canonical sequence, isoform 2 lacks residues 25–35 and 205–220; isoform 3 lacks residues 205–220. These differences could affect migration, but the features do not establish that the isoforms resolve into separate bands.
Check the antibody’s stated epitope against the listed missing regions. An epitope within residues 25–35 may be absent from isoform 2; one within 205–220 may be absent from isoforms 2 and 3. UniProt does not specify the antibody’s epitope.
Define whether the measurement covers one isoform or all detected isoforms, then use the same band selection across samples. Check antibody recognition of the regions missing from isoforms 2 and 3 before treating band intensity as total TNNT1.
Check the antibody’s stated epitope against the listed missing regions. An epitope within residues 25–35 may be absent from isoform 2; one within 205–220 may be absent from isoforms 2 and 3. UniProt does not specify the antibody’s epitope.
Define whether the measurement covers one isoform or all detected isoforms, then use the same band selection across samples. Check antibody recognition of the regions missing from isoforms 2 and 3 before treating band intensity as total TNNT1.
Could CK2 phosphorylation shift a TNNT1 band?
PTM · UniProt lists phosphoserine at position 2, modified by CK2. That position uses the supplied UniProt sequence numbering; antibody or paper numbering may differ. This feature alone does not show that phosphorylation produces a visible band shift.
Does this guide establish induction of TNNT1?
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 TNNT1?
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 A05799T1 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 TNNT1 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.
What mass should I expect for TNNT1?
Interpretation · The supplied predicted mass is 32.9 kDa; no observed band position is available. Use 32.9 kDa as a reference, not an exact migration target. The listed isoforms and phosphorylation site do not establish a visible shift or explain any measured mass difference.
Compare them with the 32.9 kDa prediction and the listed isoform differences, then check whether the antibody recognizes each isoform. UniProt also lists CK2 phosphorylation at Ser2, but none of these features alone identifies an unexpected band or proves a mobility shift.
Compare them with the 32.9 kDa prediction and the listed isoform differences, then check whether the antibody recognizes each isoform. UniProt also lists CK2 phosphorylation at Ser2, but none of these features alone identifies an unexpected band or proves a mobility shift.
Boster reagents
TNNT1 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-Troponin T1 TNNT1 Antibody
Cat # A05799T1
A05799T1 is a polyclonal anti-TNNT1 antibody listed for human, mouse, and rat reactivity. Its WB image caption reports testing at 1:500 with A549 and sp2/0 whole-cell lysates and rat skeletal muscle tissue lysate; these are the documented sample contexts.
Which to pick: A05799T1 is the only listed TNNT1 antibody and has a WB image. Check its reported 1:500 dilution and sample contexts against your experiment.
Source: BosterBio TNNT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P13805.
- Human Protein Atlas. TNNT1 tissue expression.
- PMC9938930 — target-verified WB comparison
- PMC11320084 — target-verified WB comparison
- PMC10776714 — target-verified WB comparison