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TAX1BP1 / Tax1-binding protein 1 · Western blot design guide
Design a Western Blot for TAX1BP1
Real validated TAX1BP1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TAX1BP1 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
TAX1BP1 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 | ~90.9 kDa | |
| Observed band | ~91 kDa | |
| Gel | 5–20% (catalog M04336) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| 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 C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated TAX1BP1 Western Blot Protocols
The M04336 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human RT4, human SIHA (catalog M04336) |
| Gel % | 5–20% (catalog M04336) |
| Load | 30 ug; reducing conditions (catalog M04336) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04336) |
| Membrane | nitrocellulose membrane (catalog M04336) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04336) |
| Primary antibody | M04336 · 1:500 (catalog M04336) |
| Primary incubation | overnight at 4°C (catalog M04336) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M04336) |
| Secondary incubation | 1.5 hour at RT (catalog M04336) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04336) |
| Detection | ECL (catalog M04336) |
Section 2
What Is the Expected TAX1BP1 Western Blot Band Size?
TAX1BP1 is predicted at 90.9 kDa and observed at approximately 91 kDa; no cause for the small difference is established.
| Band at approximately 91 kDa | Matches the observed band and the 90.9 kDa predicted mass; confirm identity with a target-specific control |
| Additional bands at different positions | Could reflect isoforms 1, 2, 3, or 4; distinct migration is not established |
| Doublet near 91 kDa | Could reflect differential phosphorylation, but a visible shift is not established |
| Higher-mass band | Could reflect a homooligomer if it survives sample preparation; confirm its identity |
💡Expected TAX1BP1 appearanceTAX1BP1 has a predicted mass of 90.9 kDa and an empirical band at approximately 91 kDa; confirm band identity with a target-specific control.
| UniProt predicted mass | Places the full-length protein near 90.9 kDa, consistent with the observed approximately 91 kDa band |
| Splice isoform 1 | May differ in size from other isoforms; its individual migration is not supplied |
| Splice isoform 2 | May differ in size from other isoforms; its individual migration is not supplied |
| Splice isoform 3 | May differ in size from other isoforms; its individual migration is not supplied |
| Splice isoform 4 | May differ in size from other isoforms; its individual migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A homooligomer may persist through sample preparation | Compare reducing and fully denatured samples, then confirm target identity |
| Band lower than expected | An alternative splice isoform is possible, though its migration is unknown | Check isoform recognition and confirm the band with TAX1BP1 depletion |
| Multiple bands | Isoforms or differential phosphorylation are possible; separate bands are not established | Compare isoform recognition and use TAX1BP1 depletion to identify specific bands |
| Weak or no signal | TAX1BP1 occurs in cytoplasmic and organelle-associated compartments | Check lysate recovery and compare whole-cell with relevant enriched fractions |
| Fragments below expected size | Sample degradation is possible; no signal peptide or propeptide cleavage is annotated | Prepare fresh lysate with protease inhibitors and confirm fragments with TAX1BP1 depletion |
Experiment evidence
Western blot images showing TAX1BP1 band
M04336 · TAX1BP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TAX1BP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TAX1BP1 in Western blot, you can use bone marrow lysate, which HPA scores High.
Positive control: Bone marrow (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: TAX1BP1 is intracellular, and HPA identifies not-detected tissues that can serve as negative controls.
HPA tissue expression evidence for TAX1BP1
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Ovary | follicle cells | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TAX1BP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TAX1BP1, answered from its protein features.
How should TAX1BP1 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 TAX1BP1 isoforms produce additional bands?
Isoforms · UniProt lists four isoforms. Relative to isoform 1, isoform 3 lacks residues 1–184, isoform 4 lacks 1–157, and isoforms 2–4 lack 604–645. These sequence differences could affect migration, but the features do not establish where each isoform runs.
Check the antibody epitope against the listed deletions. An epitope within isoform 1 residues 1–184 may be absent from isoform 3; one within 1–157 may be absent from isoform 4. Residues 604–645 are absent from isoforms 2–4.
Check the antibody epitope against the listed deletions. An epitope within isoform 1 residues 1–184 may be absent from isoform 3; one within 1–157 may be absent from isoform 4. Residues 604–645 are absent from isoforms 2–4.
Can phosphorylation explain a TAX1BP1 band shift?
PTM · UniProt lists phosphoserines at isoform 1 coordinates 124, 138, 225, 593, 609 and 666; residues 593 and 666 are annotated as phosphorylated by IKKA. These sites justify considering phosphorylation, but their presence does not demonstrate a visible shift. Isoform 1 coordinates should not be silently applied to shorter isoforms.
Does this guide establish induction of TAX1BP1?
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 TAX1BP1?
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 M04336 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 TAX1BP1 bands be quantified across samples?
Quantitation · Use the same defined band or band set across samples, and confirm that the antibody recognizes the isoforms being compared. TAX1BP1 is listed in cytoplasm, mitochondrion, preautophagosomal structures and autophagosomes, so compare equivalent sample fractions when measuring abundance.
Does the observed TAX1BP1 band match its predicted mass?
Interpretation · The observed band at about 91 kDa closely matches the predicted 90.9 kDa. This agreement alone does not establish which isoform is present.
Consider the listed isoform deletions and antibody epitope before assigning a band. Phosphorylation is another possibility, including IKKA-annotated sites 593 and 666 in isoform 1 coordinates. Neither feature alone identifies an unexpected band; the listed sites also do not establish a visible mobility shift.
Consider the listed isoform deletions and antibody epitope before assigning a band. Phosphorylation is another possibility, including IKKA-annotated sites 593 and 666 in isoform 1 coordinates. Neither feature alone identifies an unexpected band; the listed sites also do not establish a visible mobility shift.
Boster reagents
TAX1BP1 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-TRAF6BP Rabbit Monoclonal Antibody
Cat # M04336
The catalog reports one anti-TAX1BP1 antibody, M04336, under the TRAF6BP name. Its WB image shows an approximately 91 kDa band in human cell, rat testis, and mouse testis lysates. The supplied evidence covers these tested samples and conditions only.
Which to pick: M04336 is the only listed option. It reports human, mouse, and rat reactivity and has a WB image from HepG2, RT4, SIHA, rat testis, and mouse testis lysates. Match your sample and protocol to those reported conditions.
Source: BosterBio TAX1BP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86VP1.
- Human Protein Atlas. TAX1BP1 tissue expression.
- PMC4599966 — target-verified WB comparison
- PMC13507228 — target-verified WB comparison
- PMC10759449 — target-verified WB comparison