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- Table of Contents
TRIM29 / Tripartite motif-containing protein 29 · Western blot design guide
Design a Western Blot for TRIM29
Source-linked TRIM29 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TRIM29 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
TRIM29 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 | ~65.8 kDa | |
| Observed band | ~70 kDa | |
| Gel | 5–20% (catalog A04472-1) | |
| Positive control | Cervix (IHC candidate; verify WB) +4 more | |
| Negative control | Adrenal gland (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 | 2 isoform(s) |
Section 1
Source-Linked TRIM29 Western Blot Protocol Options
The A04472-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human RT4, human Hacat, human SIHA, human A431 (catalog A04472-1) |
| Gel % | 5–20% (catalog A04472-1) |
| Load | 30 ug; reducing conditions (catalog A04472-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04472-1) |
| Membrane | nitrocellulose membrane (catalog A04472-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04472-1) |
| Primary antibody | A04472-1 · 0.5 μg/mL (catalog A04472-1) |
| Primary incubation | overnight at 4°C (catalog A04472-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04472-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04472-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04472-1) |
| Detection | ECL (catalog A04472-1) |
Section 2
What Is the Expected TRIM29 Western Blot Band Size?
TRIM29 is calculated at 65.8 kDa and observed at ~70 kDa; the cause of this difference is not established.
| Band at ~70 kDa | Empirical TRIM29 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 65.8 kDa | Near the calculated TRIM29 mass; identity still requires controls |
| Nearby doublet | Could reflect phosphorylation, but a visible shift is unproven |
| Several bands at different positions | Could reflect Alpha and Beta isoforms, but their migration is unreported |
💡Expected TRIM29 appearanceTRIM29 has a calculated mass of 65.8 kDa and an empirical band at ~70 kDa in reducing whole-cell lysates; the cause of the difference is unknown, so confirm identity with controls.
| Calculated TRIM29 mass | 65.8 kDa is the sequence-based reference; the empirical band is ~70 kDa |
| Alpha and Beta splice isoforms | May differ in size, but their masses and separation are unreported |
| Phosphoserine at residue 21 | May affect migration; no visible shift is established |
| Phosphotyrosine at residue 106 | May affect migration; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TRIM29 occurs in the cytoplasm and lysosome; extraction or detection may be inadequate | Check lysis and compare with a TRIM29-positive whole-cell lysate |
| Band higher than expected | The reported ~70 kDa band exceeds the 65.8 kDa calculated mass for an unestablished reason | Compare with a positive lysate and confirm identity using an independent antibody or TRIM29 depletion |
| Band lower than expected | Alpha and Beta isoforms could differ in size, but their migration is unknown | Check isoform coverage and confirm the band with TRIM29 depletion |
| Multiple bands | Alpha and Beta isoforms or phosphorylation are possible; distinct bands are unproven | Confirm each band with TRIM29 depletion and, if needed, isoform-specific detection |
| Weak or no signal | TRIM29 may be poorly recovered from its cytoplasmic or lysosomal locations | Check extraction and include a TRIM29-positive lysate |
Experiment evidence
Western blot images showing TRIM29 band
A04472-1 · TRIM29
A04472 · TRIM29
M04472-2 · TRIM29
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TRIM29 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TRIM29 in Western blot, you can use cervix tissue lysate.
Positive control: Cervix (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: TRIM29 is intracellular, so tissue lysate is appropriate for detecting it.
HPA tissue expression evidence for TRIM29
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 |
|---|---|---|---|---|
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Prostate | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TRIM29 Western Blot Tips
Deeper troubleshooting and optimisation questions for TRIM29, answered from its protein features.
How should TRIM29 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 Alpha and Beta isoforms produce different bands?
Isoforms · UniProt lists Alpha and Beta. In Beta, residues 569–588 of the canonical sequence (SHYRPFYVNKGNGIGSNEAP) are replaced by VV. They may migrate differently, but separate bands are not guaranteed. Check whether the antibody epitope includes this C-terminal region.
Which reported phosphorylation sites should guide TRIM29 analysis?
PTM · UniProt lists phosphoserine at 21, 28, 58, 104 and 489; phosphotyrosine at 106; and phosphothreonine at 476. These are UniProt canonical-sequence coordinates, which may differ from paper or antibody numbering. Match any site-specific antibody to the stated numbering convention.
The seven reported phosphorylation sites make phosphorylation worth testing, but their presence does not establish a visible shift or explain the ~70 kDa band. Compare matched samples with and without phosphatase treatment if band mobility is the question.
The seven reported phosphorylation sites make phosphorylation worth testing, but their presence does not establish a visible shift or explain the ~70 kDa band. Compare matched samples with and without phosphatase treatment if band mobility is the question.
Does this guide establish induction of TRIM29?
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 TRIM29?
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 A04472-1 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 Alpha and Beta be handled during quantitation?
Quantitation · Define which band or bands are being measured and use the same choice across samples. Because Beta changes the C-terminal residues 569–588, confirm that the antibody recognizes both isoforms before interpreting a combined signal as total TRIM29.
Is a ~70 kDa band consistent with TRIM29's predicted mass?
Interpretation · TRIM29's predicted mass is 65.8 kDa, while the supplied observed band is ~70 kDa. This difference alone does not identify its cause. Check the band against a molecular weight marker and confirm its identity independently.
Consider the Alpha/Beta sequence difference and reported phosphorylation sites, then test band identity. The supplied features list no glycosylation sites, signal peptide or propeptide; they do not establish those processes as explanations for extra bands.
UniProt reports TRIM29 in the cytoplasm and lysosome, with colocalization near intermediate filaments. Record the fraction used and compare like fractions across samples; a change in one fraction's signal need not represent a change in total cellular TRIM29.
Consider the Alpha/Beta sequence difference and reported phosphorylation sites, then test band identity. The supplied features list no glycosylation sites, signal peptide or propeptide; they do not establish those processes as explanations for extra bands.
UniProt reports TRIM29 in the cytoplasm and lysosome, with colocalization near intermediate filaments. Record the fraction used and compare like fractions across samples; a change in one fraction's signal need not represent a change in total cellular TRIM29.
Boster reagents
TRIM29 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-TRIM29 Antibody Picoband®
Cat # A04472-1
Real WB data
Anti-TRIM29 Antibody (Center)
Cat # A04472
Real WB data
Anti-TRIM29 Rabbit Monoclonal Antibody
Cat # M04472-2
Three anti-TRIM29 antibodies are listed with Western blot images: A04472-1 in four human cell lysates, A04472 in SW480 lysate, and M04472-2 in JAR lysate. No publication evidence or independent validation is supplied.
Which to pick: For a human sample, choose A04472-1 if its detailed blot conditions and approximately 70 kDa band are useful; A04472 has a SW480 blot. M04472-2 lists human, mouse, and rat reactivity, but its supplied blot shows only JAR lysate.
Source: BosterBio TRIM29 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14134.
- Human Protein Atlas. TRIM29 tissue expression.
- PMC5053747 — target-verified WB comparison
- PMC7377877 — target-verified WB comparison
- PMC8340474 — target-verified WB comparison