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- Table of Contents
TAB3 / TGF-beta-activated kinase 1 and MAP3K7-binding protein 3 · Western blot design guide
Design a Western Blot for TAB3
Real validated TAB3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TAB3 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
TAB3 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 | ~78.7 kDa | |
| Observed band | ~90 kDa | |
| Gel | 5–20% (catalog A05084-3) | |
| Positive control | Adrenal gland (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated TAB3 Western Blot Protocols
The A05084-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human 293T, human Hela (catalog A05084-3) |
| Gel % | 5–20% (catalog A05084-3) |
| Load | 30 ug; reducing conditions (catalog A05084-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05084-3) |
| Membrane | nitrocellulose membrane (catalog A05084-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05084-3) |
| Primary antibody | A05084-3 · 0.5 μg/mL (catalog A05084-3) |
| Primary incubation | overnight at 4°C (catalog A05084-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05084-3) |
| Secondary incubation | 1.5 hour at RT (catalog A05084-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05084-3) |
| Detection | ECL (catalog A05084-3) |
Section 2
What Is the Expected TAB3 Western Blot Band Size?
TAB3 is predicted at 78.7 kDa and observed near 90 kDa in reducing lysates; the cause of the difference is not established.
| Band near 90 kDa | Empirical TAB3 band in reducing whole-cell lysates; confirm identity with appropriate controls. |
| Band near 78.7 kDa | Near the predicted mass; identity requires confirmation. |
| Several bands | Could reflect isoforms 1 and 2 or different modification states; distinct migration is unproven. |
| Close doublet | Could reflect phosphorylation states; a visible shift is unproven. |
💡Expected TAB3 appearanceTAB3 has a predicted mass of 78.7 kDa, while antibody QC shows a band near 90 kDa in reducing whole-cell lysates; the cause of the difference is unknown, so confirm identity with controls.
| Predicted TAB3 mass | 78.7 kDa is the sequence-based reference, not the observed migration. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Its size relative to isoform 1 and its migration are not supplied. |
| Phosphoserine sites including residues 60 and 101 | Phosphorylation may alter migration, but no visible shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The observed 90 kDa band exceeds the 78.7 kDa prediction for an undetermined reason. | Compare with the QC band and confirm identity by TAB3 depletion. |
| Band lower than expected | An isoform or fragment is possible, but its migration is not established. | Check antibody recognition and confirm the band by TAB3 depletion. |
| Multiple bands | Isoforms 1 and 2 or different phosphorylation states are possible. | Use TAB3 depletion to identify specific bands; compare phosphatase-treated samples if needed. |
| Weak or no signal | TAB3 detection or sample loading may be insufficient. | Check loading and transfer, and include a TAB3-positive control. |
| Fragments below expected size | Sample degradation is possible; no TAB3 cleavage product is specified. | Prepare fresh lysate with protease inhibitors and confirm band identity. |
Experiment evidence
Western blot images showing TAB3 band
A05084-3 · TAB3
M05084 · TAB3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TAB3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TAB3 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for TAB3
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 → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Breast | adipocytes | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced TAB3 Western Blot Tips
Deeper troubleshooting and optimisation questions for TAB3, answered from its protein features.
How should TAB3 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 TAB3 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 602–629 relative to the canonical sequence, so it has a shorter sequence. Check which isoform the antibody recognizes before assigning a second band; the feature does not establish where either isoform migrates.
Which TAB3 site is phosphorylated by MAPKAPK2 and MAPKAPK3?
PTM · UniProt annotates phosphoserine at position 506 by MAPKAPK2 and MAPKAPK3. For a site-specific assay, check that its stated residue numbering matches the UniProt canonical sequence before comparing results with antibody or paper numbering.
UniProt lists phosphoserines at positions 60, 101, 103, 385, 409 and 492, plus phosphothreonine at 404. These are UniProt canonical coordinates; antibody or paper numbering may differ. Match the assay to the intended residue and do not infer a visible mobility shift from site annotation alone.
TAB3 has multiple annotated phosphorylation sites, including serine 506 by MAPKAPK2 and MAPKAPK3. State whether the antibody measures total TAB3 or a particular phosphosite, and quantify those signals separately. A change in a site-specific signal does not by itself establish a change in total TAB3.
UniProt lists phosphoserines at positions 60, 101, 103, 385, 409 and 492, plus phosphothreonine at 404. These are UniProt canonical coordinates; antibody or paper numbering may differ. Match the assay to the intended residue and do not infer a visible mobility shift from site annotation alone.
TAB3 has multiple annotated phosphorylation sites, including serine 506 by MAPKAPK2 and MAPKAPK3. State whether the antibody measures total TAB3 or a particular phosphosite, and quantify those signals separately. A change in a site-specific signal does not by itself establish a change in total TAB3.
Does this guide establish induction of TAB3?
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 TAB3?
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 A05084-3 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 TAB3 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 TAB3 appear near 90 kDa instead of 78.7 kDa?
Interpretation · The supplied apparent band is approximately 90 kDa, while the predicted mass is 78.7 kDa. TAB3 has annotated phosphorylation and alternative splicing, but these features alone do not explain the difference or establish a visible shift. Use the 90 kDa observation as a reference and verify band identity experimentally.
UniProt annotates S-methylcysteine at position 692 under microbial infection. Consider that condition when interpreting a site-specific result, and confirm whether any experimental condition involved infection. The annotation does not show that this modification creates a separate Western-blot band.
Compare it with the approximately 90 kDa observed band and the 78.7 kDa predicted mass. Check whether the antibody recognizes isoform 2, which lacks canonical residues 602–629. TAB3 also has N-acetylalanine at position 2 and annotated phosphorylation, but their presence alone does not identify the unexpected band.
UniProt annotates S-methylcysteine at position 692 under microbial infection. Consider that condition when interpreting a site-specific result, and confirm whether any experimental condition involved infection. The annotation does not show that this modification creates a separate Western-blot band.
Compare it with the approximately 90 kDa observed band and the 78.7 kDa predicted mass. Check whether the antibody recognizes isoform 2, which lacks canonical residues 602–629. TAB3 also has N-acetylalanine at position 2 and annotated phosphorylation, but their presence alone does not identify the unexpected band.
Boster reagents
TAB3 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-TAB3 Antibody Picoband®
Cat # A05084-3
Real WB data
Anti-TAB3 Rabbit Monoclonal Antibody
Cat # M05084
The catalog reports two anti-TAB3 antibodies with Western blot images. A05084-3 was tested in human cell lysates; M05084 in human and mouse cell lysates. Both captions report a band near 90 kDa, above the expected 79 kDa; no further validation is supplied.
Which to pick: For human lysates, either A05084-3 or M05084 has a Western blot image. For mouse lysates, choose M05084, whose image includes Raw264.7 and NIH/3T3 samples. Match your sample and conditions to the reported tests.
Source: BosterBio TAB3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8N5C8.
- Human Protein Atlas. TAB3 tissue expression.
- PMC10236263 — target-verified WB comparison
- PMC5242086 — target-verified WB comparison
- PMC11648720 — target-verified WB comparison