This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TRADD / Tumor necrosis factor receptor type 1-associated DEATH domain protein · Western blot design guide
Design a Western Blot for TRADD
Real validated TRADD Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TRADD 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
TRADD 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 | ~34.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Esophagus (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Microbial infection-dependent glycosylation | |
| Caveat | Infection-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated TRADD Western Blot Protocols
The M02785 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Hela cell lysate (catalog M02785) |
| 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 | M02785; 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 TRADD Western Blot Band Size?
TRADD is predicted at 34.2 kDa; splice isoforms and infection-associated Arg235/Arg245 GlcNAc are annotated, but their effects on migration are unproven.
| Band near 34.2 kDa | Consistent with the predicted TRADD mass; confirm identity with controls. |
| Two bands at different positions | Isoforms 1 and 2 are possible contributors, but their migration is not established. |
| No visible shift after microbial infection | The annotated GlcNAc modifications at Arg235 and Arg245 need not produce a resolvable shift. |
| Infection-associated mobility change | Modification at Arg235 or Arg245 is possible; confirm the band identity and modification. |
💡Expected TRADD appearanceTRADD has a predicted mass of 34.2 kDa, but no empirical band size is supplied; use TRADD depletion and suitable antibody controls to identify any detected band.
| Predicted TRADD mass | 34.2 kDa is the sequence-based reference, not a measured band position. |
| Microbial infection-associated GlcNAc at Arg235 | May affect mobility if present, but a visible shift is not established. |
| Microbial infection-associated GlcNAc at Arg245 | May affect mobility if present, but a visible shift is not established. |
| Splice isoforms 1 and 2 | May differ in size; their individual masses and band positions are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TRADD may be missed if extraction does not recover its nuclear or cytoplasmic pool. | Check recovery from both compartments and include an antibody control. |
| Band higher than expected | The band identity or migration relative to the 34.2 kDa prediction is unconfirmed. | Check the size marker and test whether TRADD depletion removes the band. |
| Band lower than expected | An alternative isoform is possible, but its size is unknown. | Confirm band identity by TRADD depletion and compare isoform expression. |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are not established. | Check which bands respond to TRADD depletion and assess isoform expression. |
| Weak or no signal | Nuclear and cytoplasmic localization may affect recovery from a fractionated sample. | Check both fractions, loading, and an antibody control. |
Experiment evidence
Western blot images showing TRADD band
M02785 · TRADD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TRADD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TRADD in Western blot, you can use adrenal gland lysate, based on HPA detection.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Esophagus (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA rates adrenal gland expression as medium, so the positive signal may be modest.
HPA tissue expression evidence for TRADD
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Skin | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced TRADD Western Blot Tips
Deeper troubleshooting and optimisation questions for TRADD, answered from its protein features.
How should TRADD 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 TRADD isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–60 of the canonical sequence and may migrate differently. Check whether the antibody recognizes that N-terminal region before assigning bands to either isoform.
Can infection-associated glycosylation affect TRADD bands?
PTM · UniProt lists N-beta-linked GlcNAc on arginine at positions 235 and 245 under microbial infection conditions. These are UniProt canonical coordinates; antibody or paper numbering may differ. Compare infected and control samples if investigating these modifications. Their presence in the feature list does not establish a visible band shift.
Does TNF receptor stimulation increase TRADD abundance?
Induction · The supplied feature says TNFRSF1A stimulation leads to formation of distinct signaling complexes. It does not establish increased TRADD abundance. If testing stimulation, quantify TRADD against an appropriate loading reference and interpret changes separately from complex formation.
How should transfer be checked for TRADD?
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 M02785 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 I quantify TRADD across cell fractions?
Quantitation · TRADD is listed in the cytoplasm, cytoskeleton and nucleus, and shuttles between cytoplasm and nucleus. Quantify comparable fractions with appropriate fraction-specific loading references. A change in one fraction may reflect redistribution rather than a change in total TRADD.
What mass should I expect for TRADD?
Interpretation · The predicted mass is 34.2 kDa for canonical TRADD. No observed band position is supplied, so use 34.2 kDa as a reference, not a confirmed apparent mass. The listed features alone cannot explain a difference between calculated and observed mass.
Consider the two listed isoforms, including isoform 2’s deletion of canonical residues 1–60. Infection-associated GlcNAc modifications are listed at canonical arginine positions 235 and 245, but they do not establish a visible shift. Compare relevant sample conditions and antibody epitope coverage before assigning an unexpected band.
Consider the two listed isoforms, including isoform 2’s deletion of canonical residues 1–60. Infection-associated GlcNAc modifications are listed at canonical arginine positions 235 and 245, but they do not establish a visible shift. Compare relevant sample conditions and antibody epitope coverage before assigning an unexpected band.
Boster reagents
TRADD 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-TRADD Monoclonal Antibody
Cat # M02785
The catalog reports one anti-TRADD antibody for Western blot: M02785, a monoclonal antibody with stated human, mouse, and rat reactivity. Its supplied WB image shows TRADD expression in HeLa cell lysate; no other sample validation is provided.
Which to pick: M02785 is the only listed option. Its WB image uses HeLa cell lysate, making that the documented test context. Human, mouse, and rat reactivity is listed, but the supplied image does not demonstrate WB performance in mouse or rat samples.
Source: BosterBio TRADD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15628.
- Human Protein Atlas. TRADD tissue expression.
- PMC10549964 — target-verified WB comparison
- PMC4768865 — target-verified WB comparison
- PMC3078067 — target-verified WB comparison