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- Table of Contents
YWHAB / 14-3-3 protein beta/alpha · Western blot design guide
Design a Western Blot for YWHAB
Source-linked YWHAB Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-YWHAB 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
YWHAB 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 | ~28.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Heart muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked YWHAB Western Blot Protocol Options
The M02431-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Hela lysate (catalog M02431-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02431-1; 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 YWHAB Western Blot Band Size?
YWHAB has a predicted 28.1 kDa monomer; its homodimer, isoforms, and modifications could affect bands, but no empirical migration is supplied.
| Band near 28.1 kDa | consistent with the predicted YWHAB monomer; confirm identity with controls |
| Band near 56 kDa | could reflect the annotated homodimer if it survives sample preparation |
| Two bands near the monomer region | could reflect Long and Short isoforms; their separation is unestablished |
| Band slightly offset from 28.1 kDa | N-terminal processing or listed modifications are possible, but their migration effects are unestablished |
💡Expected YWHAB appearanceThe predicted YWHAB monomer is 28.1 kDa; no empirical band size is supplied, so confirm any band with identity controls before assigning an isoform or modification.
| Predicted monomer mass | sets a reference of 28.1 kDa, not a measured migration position |
| Homodimer | could appear near twice the monomer size if preserved during electrophoresis |
| Long isoform | may migrate differently from Short; no isoform mass is supplied |
| Short isoform | may migrate differently from Long; no isoform mass is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytoplasmic YWHAB may be lost during fractionation | check the soluble cytoplasmic fraction and a positive lysate control |
| Band higher than expected | the annotated homodimer may have survived sample preparation | compare fully denatured and less denatured samples and confirm band identity |
| Band lower than expected | the Short isoform or protein breakdown may account for the band | compare isoform controls and freshly prepared lysate |
| Multiple bands | Long and Short isoforms or listed modifications may contribute | use isoform controls and an independent YWHAB antibody to identify the bands |
| Fragments below expected size | YWHAB may have degraded during lysate preparation | prepare fresh lysate with protease inhibitors and compare band identity |
Experiment evidence
Western blot images showing YWHAB band
M02431-1 · YWHAB
M02431-2 · YWHAB
M02431-3 · YWHAB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for YWHAB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for YWHAB in Western blot, you can use caudate tissue, which has high HPA expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Heart muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Tissue controls are feasible because HPA reports high expression in caudate and no detection in heart muscle.
HPA tissue expression evidence for YWHAB
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 |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced YWHAB Western Blot Tips
Deeper troubleshooting and optimisation questions for YWHAB, answered from its protein features.
Where should the YWHAB band appear?
Band shift · The predicted mass is 28.1 kDa. No empirical apparent band is supplied, so use 28.1 kDa as a reference rather than an expected observed position. The listed modifications alone do not establish a visible shift.
Can the Long and Short isoforms be resolved by Western blot?
Isoforms · UniProt lists Long and Short isoforms; Short lacks residues 1–2 of the canonical sequence. Their small sequence difference does not establish that they will resolve as separate bands. Check which isoform the antibody recognizes before assigning bands.
Which phosphorylation sites matter when interpreting YWHAB bands?
PTM · UniProt lists phosphothreonine at position 2 and phosphoserine at positions 60 and 186. Its position 232 modification entry reads ‘Phosphoserine N-acetylmethionine.’ These are UniProt coordinates; check the numbering convention used by the antibody. Site annotations alone cannot identify a shifted band.
UniProt lists N-terminal acetylation at position 1 or, after N-terminal processing, position 2; lysine acetylation at positions 5, 51, 70, and 117; and tyrosine nitration at positions 84 and 106. Compare antibody epitope information with these UniProt coordinates when signal varies. The annotations do not establish an effect on binding.
UniProt lists N-terminal acetylation at position 1 or, after N-terminal processing, position 2; lysine acetylation at positions 5, 51, 70, and 117; and tyrosine nitration at positions 84 and 106. Compare antibody epitope information with these UniProt coordinates when signal varies. The annotations do not establish an effect on binding.
Does this guide establish induction of YWHAB?
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 YWHAB?
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 M02431-1 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 YWHAB be quantified across samples?
Quantitation · Quantify a consistently identified YWHAB band using the same antibody and sample preparation across samples. UniProt places YWHAB in the cytoplasm and reports it in melanosome fractions, so keep the sampled fraction consistent. The feature list alone cannot establish whether multiple bands represent equivalent amounts of YWHAB.
Does a higher band indicate a YWHAB dimer?
Interpretation · UniProt describes YWHAB as a homodimer. That makes dimerization relevant when investigating a higher band, but the feature does not establish that a dimer survives sample preparation or causes that band. Do not assign its identity from position alone.
Start with the 28.1 kDa predicted mass and the Long/Short difference of two N-terminal residues. UniProt also lists several modifications, but none proves a visible shift or explains an observed mass difference. Confirm band identity before attributing it to an isoform or modification.
Start with the 28.1 kDa predicted mass and the Long/Short difference of two N-terminal residues. UniProt also lists several modifications, but none proves a visible shift or explains an observed mass difference. Confirm band identity before attributing it to an isoform or modification.
Boster reagents
YWHAB 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-14-3-3 YWHAB Rabbit Monoclonal Antibody
Cat # M02431-1
Real WB data
Anti-14-3-3 YWHAB Rabbit Monoclonal Antibody
Cat # M02431-2
Real WB data
Anti-14-3-3 alpha + beta YWHAB Rabbit Monoclonal Antibody
Cat # M02431-3
Three the supplier rabbit monoclonal anti-YWHAB antibodies list human, mouse, and rat reactivity and have Western blot images. Captions document specific cell or tissue lysates; no publication evidence is supplied. These examples do not establish performance in every sample type.
Which to pick: Choose M02431-3 for the most detailed WB example: human cell and rat and mouse tissue lysates, with a reported 28 kDa band. M02431-2 shows HeLa, 3T3, and PC-12 lysates; M02431-1 shows HeLa lysate. Match your sample to the documented context.
Source: BosterBio YWHAB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P31946.
- Human Protein Atlas. YWHAB tissue expression.
- PMC7568206 — target-verified WB comparison
- PMC7850404 — target-verified WB comparison
- PMC10296083 — target-verified WB comparison