YWHAQ / 14-3-3 protein theta · Western blot design guide

Design a Western Blot for YWHAQ

Source-linked YWHAQ Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-YWHAQ 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

Western blot protocol sheet for YWHAQ: expected band ~27.8 kDa, hero antibody M03904, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **YWHAQ** Western blot protocol sheet — expected band ~27.8 kDa, antibody M03904, controls and PMC citations. Open the full YWHAQ WB guide →

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YWHAQ 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	~27.8 kDa
	Observed band	~28 kDa
	Gel	12% (catalog M03904)
	Positive control ⓘ	Colon (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 + Acetylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Source-Linked YWHAQ Western Blot Protocol Options

The M03904 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human Hela, human A549, human SH-SY5Y, human U251, rat brain, rat C6, mouse brain (catalog M03904)
Gel %	12% (catalog M03904)
Load	30 ug; reducing conditions (catalog M03904)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03904)
Membrane	nitrocellulose membrane (catalog M03904)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M03904)
Primary antibody	M03904 · 1:5000 (catalog M03904)
Primary incubation	overnight at 4°C (catalog M03904)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog M03904)
Secondary incubation	1.5 hour at RT (catalog M03904)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M03904)
Detection	ECL (catalog M03904)

Section 2

What Is the Expected YWHAQ Western Blot Band Size?

YWHAQ is predicted at 27.8 kDa and observed near 28 kDa; this small difference has no established cause in the supplied evidence.

What am I looking at on my blot?
Band near 28 kDa	Matches the empirical YWHAQ band and its 27.8 kDa predicted monomer mass.
Band near 56 kDa	Could represent a retained YWHAQ homodimer; confirm its identity experimentally.
Close doublet near 28 kDa	Could reflect a modified species, but the annotated modifications do not establish a visible shift.
Band near 28 kDa in whole-cell lysate	Consistent with cytoplasmic YWHAQ detected in the supplied lysate blots.

💡Expected YWHAQ appearanceYWHAQ has a predicted monomer mass of 27.8 kDa and an empirical band near 28 kDa; confirm band identity with appropriate antibody controls.

How each factor affects band size
UniProt predicted monomer mass	Predicts a band near 27.8 kDa.
245-residue YWHAQ polypeptide	Provides the sequence basis for the predicted monomer mass.
YWHAQ homodimer	Could appear near twice the monomer mass if the dimer persists during electrophoresis.
Homodimer dissociation	Leaves monomeric YWHAQ near the predicted mass.

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Cytoplasmic YWHAQ may be below detection in the sample.	Check lysate loading and use a lysate with a documented 28 kDa signal.
Band higher than expected	A retained homodimer could migrate near twice the monomer mass.	Compare preparation conditions and confirm YWHAQ identity with an independent antibody.
Band lower than expected	A smaller band has no established YWHAQ processing explanation in the supplied features.	Check band identity with an independent antibody and inspect sample handling.
Multiple bands	Annotated modifications or a retained homodimer are possible, but their migration effects are unproven.	Compare sample preparation conditions and verify each band's identity.
Weak or no signal	YWHAQ abundance or assay sensitivity may be insufficient.	Check transfer and antibody conditions against a documented positive lysate.

Source: UniProt P27348 Human Protein Atlas product datasheet (M03904)

Experiment evidence

Western blot images showing YWHAQ band

M03904 · YWHAQ

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for YWHAQ Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for YWHAQ in Western blot, you can use colon tissue lysate.

Positive control: Colon (IHC candidate; verify WB)

Negative control: Adipose tissue (IHC candidate; verify WB)

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: A colon lysate is a feasible positive control for cytoplasmic YWHAQ; adipose tissue is a candidate negative control based on HPA non-detection.

HPA tissue expression evidence for YWHAQ

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
Colon	peripheral nerve/ganglion	High	Protein (IHC)	HPA →
Adrenal gland	glandular cells	Medium	Protein (IHC)	HPA →
Appendix	glandular cells	Medium	Protein (IHC)	HPA →
Breast	glandular 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	Not detected	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	Not detected	Protein (IHC)	HPA →
Heart muscle	cardiomyocytes	Not detected	Protein (IHC)	HPA →
Skeletal muscle	myocytes	Not detected	Protein (IHC)	HPA →
Smooth muscle	smooth muscle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced YWHAQ Western Blot Tips

Deeper troubleshooting and optimisation questions for YWHAQ, answered from its protein features.

How should YWHAQ 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 YWHAQ isoforms explain multiple bands?

Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign additional bands to YWHAQ isoforms on this evidence alone.

Which phosphorylation sites matter when interpreting YWHAQ bands?

PTM · UniProt lists phosphoserine at positions 92 and 232. If testing phosphorylation, compare site-specific and total YWHAQ signals. These are UniProt coordinates; antibody or paper numbering may differ. The sites do not establish a visible band shift.

UniProt lists N-acetylmethionine at position 1 and N6-acetyllysine at positions 3, 49 (alternate), 68, and 115. Check antibody specificity before interpreting a modified-protein signal. These are UniProt coordinates and do not by themselves explain a mass difference.

Does this guide establish induction of YWHAQ?

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 YWHAQ?

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 M03904 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 total YWHAQ be quantified alongside modified YWHAQ?

Quantitation · Measure the approximately 28 kDa total YWHAQ band consistently across samples, and assess modification-specific signals separately. Because phosphorylation, acetylation, and nitration are listed, a change in a modification-specific signal alone does not establish a change in total YWHAQ abundance.

Should YWHAQ migrate near its predicted mass?

Interpretation · The predicted mass is 27.8 kDa, and the supplied observed band is approximately 28 kDa. These values agree closely. The listed modifications alone do not establish a visible shift.

UniProt lists 3'-nitrotyrosine at positions 82 and 104. A nitration-specific signal should be interpreted separately from total YWHAQ. These UniProt coordinates may differ from antibody or paper numbering; the listed sites do not establish a visible shift.

YWHAQ is annotated as a homodimer, but that annotation alone does not identify a higher Western-blot band. Compare it with the approximately 28 kDa observed band and verify its identity before attributing it to dimerization or a listed modification.

Boster reagents

YWHAQ 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

Western blot analysis of YWHAQ using anti-YWHAQ antibody (M03904). Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat C6 whole cell lysates, Lane 7: mouse brain tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-YWHAQ antigen affinity purified monoclonal antibody (M03904) at 1:5000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for YWHAQ at approximately 28 kDa. The expected band size for YWHAQ is at 28 kDa.

Anti-14-3-3 Theta YWHAQ Rabbit Monoclonal Antibody

Cat # M03904

View product →

The catalog reports one anti-YWHAQ antibody, M03904, with reported human, mouse, and rat reactivity. Its Western blot image shows an approximately 28 kDa band in the listed cell and brain lysates. No independent publication or orthogonal validation evidence is supplied.

Which to pick: M03904 is the only listed option. Its WB image includes human HeLa, A549, SH-SY5Y, and U251 cells; rat brain and C6 cells; and mouse brain. Use those tested samples and the reported 1:5000 primary dilution to assess fit for your experiment.

Source: BosterBio YWHAQ gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry P27348.
Human Protein Atlas. YWHAQ tissue expression.
PMC3174057 — target-verified WB comparison