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- Table of Contents
TARDBP / TAR DNA-binding protein 43 · Western blot design guide
Design a Western Blot for TARDBP
Real validated TARDBP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TARDBP 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
TARDBP 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 | ~44.7 kDa | |
| Observed band | ~43 kDa | |
| Gel | 5–20% (catalog A01001-3) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Methylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated TARDBP Western Blot Protocols
The A01001-3 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, human HepG2 (catalog A01001-3) |
| Gel % | 5–20% (catalog A01001-3) |
| Load | 30 ug; reducing conditions (catalog A01001-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01001-3) |
| Membrane | nitrocellulose membrane (catalog A01001-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01001-3) |
| Primary antibody | A01001-3 · 0.5 μg/mL (catalog A01001-3) |
| Primary incubation | overnight at 4°C (catalog A01001-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01001-3) |
| Secondary incubation | 1.5 hour at RT (catalog A01001-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01001-3) |
| Detection | ECL (catalog A01001-3) |
Section 2
What Is the Expected TARDBP Western Blot Band Size?
TARDBP is predicted at 44.7 kDa and observed at ~43 kDa; the cause of the difference is not established.
| Band at ~43 kDa | Empirical TARDBP band; predicted mass is 44.7 kDa |
| Band near twice the monomer size | Possible homodimer if it persists during electrophoresis |
| Higher-mass bands | Possible N-terminal-domain-mediated homooligomers if they persist during electrophoresis |
| Several bands at different positions | Could reflect isoforms 1 and 2 if their migration differs enough to resolve |
💡Expected TARDBP appearanceTARDBP has a predicted mass of 44.7 kDa and an observed band at ~43 kDa; the difference is unexplained, so confirm identity with ordinary antibody controls.
| Predicted TARDBP mass | 44.7 kDa is the sequence-based reference; the observed band is ~43 kDa |
| Homodimer formation | Could yield a band near twice the monomer size if the dimer persists during electrophoresis |
| N-terminal-domain-mediated homooligomer formation | Could yield higher-mass bands if oligomers persist during electrophoresis |
| Isoform 1 | Its individual mass and migration relative to isoform 2 are not supplied |
| Isoform 2 | Its individual mass and migration relative to isoform 1 are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TARDBP may be poorly recovered from the nucleus | Check nuclear extraction and include a positive-control lysate |
| Band higher than expected | TARDBP homodimers or homooligomers may persist during electrophoresis | Compare sample denaturation conditions and validate the band with antibody controls |
| Band lower than expected | The observed ~43 kDa band is below the 44.7 kDa prediction for an unestablished reason | Compare with a positive-control lysate and verify antibody specificity |
| Multiple bands | Isoforms 1 and 2 or persistent oligomers are possible, but distinct migration is not established | Compare denaturation conditions and use band-identity controls |
| Weak or no signal | A sampled fraction may contain little TARDBP because it occurs in the nucleus and cytoplasm | Check both fractions and include a positive-control lysate |
Experiment evidence
Western blot images showing TARDBP band
A01001-3 · TARDBP
M01001-2 · TARDBP
M01001 · TARDBP
A01001 · TARDBP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TARDBP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TARDBP in Western blot, you can use adipose tissue, an HPA high-expression sample.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No clean negative tissue is shown; use siRNA knockdown or a KO line.
HPA tissue expression evidence for TARDBP
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Medium | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Medium | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced TARDBP Western Blot Tips
Deeper troubleshooting and optimisation questions for TARDBP, answered from its protein features.
How should TARDBP 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.
How could isoform 2 affect band identification?
Isoforms · Isoform 2 replaces residues 1–18 and lacks residues 19–134 relative to the canonical sequence. It may therefore appear smaller. An antibody targeting the missing region would not detect isoform 2; check the antibody epitope before interpreting a lower band.
Which TARDBP modifications matter when comparing bands?
PTM · UniProt lists phosphoserines at positions 183 and 292 and omega-N-methylarginine at position 293. These are UniProt canonical-sequence coordinates, which may differ from paper or antibody numbering. Their presence alone does not establish a visible band shift.
Does this guide establish induction of TARDBP?
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 TARDBP?
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 A01001-3 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 TARDBP 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.
Why might TARDBP appear near 43 kDa instead of 44.7 kDa?
Interpretation · The supplied apparent band is about 43 kDa, while the predicted mass is 44.7 kDa. These values differ modestly; the listed features do not establish the cause. Identify bands using an appropriate TARDBP control rather than assigning the difference to a specific modification.
TARDBP is listed in both the nucleus and cytoplasm. When comparing localization, analyze the fractions consistently and quantify the TARDBP signal within each fraction. A change in one fraction alone does not establish a change in total TARDBP.
Consider isoform 2, which lacks canonical residues 19–134 and has a different sequence at residues 1–18. Check whether the antibody recognizes a region retained in isoform 2. The supplied features alone cannot identify a particular unexpected band.
TARDBP is reported as a homodimer and as a homooligomer through its N-terminal domain. Those features make oligomerization relevant when investigating higher bands, but they do not prove that an observed Western-blot band is an oligomer. Verify its identity experimentally.
TARDBP is listed in both the nucleus and cytoplasm. When comparing localization, analyze the fractions consistently and quantify the TARDBP signal within each fraction. A change in one fraction alone does not establish a change in total TARDBP.
Consider isoform 2, which lacks canonical residues 19–134 and has a different sequence at residues 1–18. Check whether the antibody recognizes a region retained in isoform 2. The supplied features alone cannot identify a particular unexpected band.
TARDBP is reported as a homodimer and as a homooligomer through its N-terminal domain. Those features make oligomerization relevant when investigating higher bands, but they do not prove that an observed Western-blot band is an oligomer. Verify its identity experimentally.
Boster reagents
TARDBP 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-TDP-43/TARDBP Antibody Picoband®
Cat # A01001-3
Real WB data
Anti-TDP43 Rabbit Monoclonal Antibody
Cat # M01001-2
Real WB data
Anti-TDP43 TARDBP Rabbit Monoclonal Antibody
Cat # M01001
Real WB data
Anti-TDP43 TARDBP Antibody
Cat # A01001
Four the supplier anti-TARDBP antibodies have WB images. Their captions show human, mouse, or rat lysates; reported bands for A01001-3 and M01001 are approximately 43 and 40 kDa, respectively, versus an expected 45 kDa. The supplied evidence does not establish specificity independently.
Which to pick: Match the image to your sample: M01001-2 shows HeLa, mouse brain, and rat brain; M01001 shows additional human cell lines and rodent samples. A01001-3 shows human, rat, and mouse lysates; A01001 shows HeLa at three concentrations. M01001 lists zebrafish reactivity, but its WB image has no zebrafish sample.
Source: BosterBio TARDBP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13148.
- Human Protein Atlas. TARDBP tissue expression.
- PMC13455517 — target-verified WB comparison
- PMC13180789 — target-verified WB comparison
- PMC5289002 — target-verified WB comparison