TST / Thiosulfate sulfurtransferase · Western blot design guide

Design a Western Blot for TST

Real validated TST Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TST 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 TST: expected band ~33.4 kDa, hero antibody A00965-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **TST** Western blot protocol sheet — expected band ~33.4 kDa, antibody A00965-1, controls and PMC citations. Open the full TST WB guide →

Navigate to: Protocols Band Controls Antibodies

TST 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	~33.4 kDa
	Observed band	~34 kDa
	Gel	5–20% (catalog A00965-1)
	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	Glycosylated + Phosphorylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Real Curated TST Western Blot Protocols

The A00965-1 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 293T, human HepG2, human MCF-7 (catalog A00965-1)
Gel %	5–20% (catalog A00965-1)
Load	30 ug; reducing conditions (catalog A00965-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00965-1)
Membrane	nitrocellulose membrane (catalog A00965-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A00965-1)
Primary antibody	A00965-1 · 0.5 μg/mL (catalog A00965-1)
Primary incubation	overnight at 4°C (catalog A00965-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A00965-1)
Secondary incubation	1.5 hour at RT (catalog A00965-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A00965-1)
Detection	ECL (catalog A00965-1)

Section 2

What Is the Expected TST Western Blot Band Size?

TST is predicted at 33.4 kDa and observed at ~34 kDa; the cause of this small difference is not established.

What am I looking at on my blot?
Band at ~34 kDa	Empirical TST band, close to its 33.4 kDa predicted mass.
Single sharp band near 33.4 kDa	Consistent with the predicted size of monomeric TST.
Weak or absent band in whole-cell lysate	Mitochondrial matrix localization may limit the detectable signal.
No separate band attributable to glycosylation	Ser35 O-linked GlcNAc does not establish a resolvable mobility shift.

💡Expected TST appearanceTST has a predicted mass of 33.4 kDa and an empirical band at ~34 kDa; confirm band identity with antibody and sample controls because the supplied features do not establish the cause of the small difference.

How each factor affects band size
Predicted monomer mass	33.4 kDa predicted; the empirical TST band is ~34 kDa.
Ser35 O-linked GlcNAc	A single listed site does not establish a visible size shift.
Ser38 phosphorylation	May affect mobility, but no distinct band is established.
Lys163 acetylation	No resolvable size effect is established.
Lys14 alternate acetylation or succinylation	No distinct migration pattern is established.

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	TST is localized to the mitochondrial matrix.	Check a mitochondria-enriched fraction and a mitochondrial marker.
Band higher than expected	The listed modifications do not establish the cause of a substantial shift.	Compare with the ~34 kDa reference and verify identity using an independent antibody or TST depletion.
Band lower than expected	A smaller band has no established processing explanation in the supplied features.	Check sample integrity and verify identity using TST depletion.
Broad smear instead of sharp band	The single Ser35 GlcNAc site does not establish a smear.	Check sample quality and loading, then compare with a positive-control lysate.
Multiple bands	Additional bands are not explained by named isoforms or disulfide-linked TST.	Use TST depletion or an independent antibody to identify the specific band.
Weak or no signal	Whole-cell samples may have limited detectable mitochondrial TST.	Check loading and transfer, then test a mitochondria-enriched sample.
Fragments below expected size	Sample degradation is possible; no cleavage product is specified.	Prepare fresh lysate with protease inhibitors and verify band identity.

Source: UniProt Q16762 Human Protein Atlas product datasheet (A00965-1)

Experiment evidence

Western blot images showing TST band

A00965-1 · TST

← swipe / scroll for more images →

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

Sample controls for TST Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TST in Western blot, you can use adrenal gland tissue, which HPA rates high for TST.

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 such as Ponceau alongside TST.

⚠️Feasibility: No HPA not-detected tissue is listed, so a knockdown or KO sample is the clearest negative control.

HPA tissue expression evidence for TST

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	High	Protein (IHC)	HPA →
Appendix	glandular cells	High	Protein (IHC)	HPA →
Bronchus	respiratory epithelial 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
Bone marrow	hematopoietic cells	Low	Protein (IHC)	HPA →
Epididymis	glandular cells	Low	Protein (IHC)	HPA →
Esophagus	squamous epithelial cells	Low	Protein (IHC)	HPA →
Heart muscle	cardiomyocytes	Low	Protein (IHC)	HPA →
Hippocampus	glial cells	Low	Protein (IHC)	HPA →

Section 3

Advanced TST Western Blot Tips

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

How should TST 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 TST isoforms explain multiple bands?

Isoforms · Only one isoform is supplied, with no alternative sequence. These features do not support assigning additional bands to TST isoforms.

Which TST glycosylation site is relevant to antibody recognition?

PTM · UniProt lists O-linked GlcNAc at serine 35. If antibody binding varies between samples, check whether its epitope includes this region. UniProt coordinates may differ from antibody or paper numbering.

UniProt lists phosphoserine 38. Check whether an antibody epitope spans this site when comparing signals. The listed site alone does not establish a separate Western blot band.

UniProt lists acetylation at lysines 14, 136, 163, 175, 224, 236 and 237; succinylation is listed as an alternate modification at 14, 136, 175, 224 and 237. Compare the antibody epitope with these UniProt coordinates. These annotations do not establish distinct bands.

Does this guide establish induction of TST?

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 TST?

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 A00965-1 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 TST 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 TST run near 34 kDa rather than 33.4 kDa?

Interpretation · The observed band is approximately 34 kDa, close to the predicted 33.4 kDa. TST has listed modifications, but their presence alone does not establish a visible shift or explain the small difference in apparent mass.

TST is listed in the mitochondrial matrix. Use comparable sample preparation and loading when quantifying the band near 34 kDa; differences in mitochondrial content could affect its signal.

Start with the approximately 34 kDa observed band and the 33.4 kDa predicted mass. TST is listed as a monomer, with one isoform and no annotated signal peptide or propeptide. Its modifications alone do not identify an unexpected band; verify antibody specificity before assigning it to TST.

Boster reagents

TST 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 TST using anti-TST antibody (A00965-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human 293T whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: rat kidney tissue lysates, Lane 5: rat liver tissue lysates, Lane 6: mouse liver 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-TST antigen affinity purified polyclonal antibody (Catalog # A00965-1) at 0.5 μg/mL 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TST at approximately 34 kDa. The expected band size for TST is at 33 kDa.

Anti-TST Antibody Picoband®

Cat # A00965-1

View product →

A00965-1 is listed for human, mouse, and rat TST. Its Western blot image reports an approximately 34 kDa band, compared with an expected 33 kDa, in human cell, rat tissue, and mouse liver lysates. No publication evidence was supplied.

Which to pick: A00965-1 is the only listed antibody. It has a Western blot image using human 293T, HepG2, and MCF-7 cells; rat kidney and liver; and mouse liver. Match your sample to these tested contexts when choosing it.

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

References

UniProt Consortium. UniProt entry Q16762.
Human Protein Atlas. TST tissue expression.
PMC12664448 — target-verified WB comparison
PMC5567475 — target-verified WB comparison
PMC9947110 — target-verified WB comparison