SMTN · Western blot design guide

SMTN Western Blot Planning Guide

Plan a SMTN Western blot around the catalog-observed 99.1 kDa band, image-backed A04895 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use

Western blot protocol sheet for SMTN (SMTN): expected band 99.1 kDa, antibody A04895, and PMC-cited SDS-PAGE protocol steps — SMTN Western blot protocol sheet — expected band 99.1 kDa, antibody A04895, controls and PMC citations. Open the full SMTN WB guide →

Navigate to: Protocols Band Controls Antibodies

SMTN 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	99.1 kDa
	Observed band	Not reported — verify product WB image
	Gel	8-10%
	Positive control ⓘ	Smooth muscle
	Negative control ⓘ	Appendix

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	99.1 kDa
ⓘ	Localization	Cytoplasm, cytoskeleton
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Section 1

Real Curated SMTN Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysate	Smooth muscle
Gel %	8-10%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, standard transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A04895 at datasheet starting dilution
Primary incubation	Overnight at 4 °C with gentle agitation
Secondary	Species-matched HRP conjugate at validated dilution
Wash	3 × 5 min in TBST
Detection	Chemiluminescent substrate
Exposure	Bracket exposures to avoid saturation

Section 2

What Is the Expected SMTN Western Blot Band Size?

Use the product-observed 99.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
99.1 kDa	Matches the authoritative product WB observation.
99.1 kDa calculated	Use as UniProt context, not as a replacement observed band.
Unexpected additional signal	Do not assign identity without orthogonal positive/negative controls.

💡Expected SMTN appearancePlan around 99.1 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	99.1 kDa; use this as the primary experimental expectation.
Calculated mass	99.1 kDa from UniProt P53814; retain as context.
Gel selection	8-10%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A04895.

Why is my band missing or off?
Situation	Likely cause	Next action
99.1 kDa	Matches the authoritative product WB observation.	Confirm with orthogonal controls and the linked product record.
Additional band	May reflect processing, modification, or non-specific signal.	Run a dilution series and compare positive/negative controls.
Weak signal	Target abundance or transfer may be limiting.	Verify transfer, increase positive-control abundance, and bracket exposure.

Sources: UniProt P53814 Human Protein Atlas product datasheet (A04895)

Experiment evidence

Western blot images showing SMTN band

A04895 · SMTN

← swipe / scroll for more images →

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

Sample controls for SMTN Western blot

🧪Use Smooth muscle as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.

Positive control: Smooth muscle (High)

Negative control: Appendix (Not detected)

HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for SMTN

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.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Smooth muscle	Reported tissue cells	High	Protein (HPA)	HPA →
Testis	Reported tissue cells	Medium	Protein (HPA)	HPA →
Kidney	Reported tissue cells	Medium	Protein (HPA)	HPA →
Heart muscle	Reported tissue cells	Medium	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Appendix	Reported tissue cells	Not detected	Protein (HPA)	HPA →
Bone marrow	Reported tissue cells	Not detected	Protein (HPA)	HPA →

Section 3

Advanced SMTN Western Blot Tips

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

Which band should guide the blot?

Use 99.1 kDa, the observation attached to the authoritative A04895 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 99.1 kDa expectation.

Which positive control should I start with?

Start with Smooth muscle, the lead HPA protein-expression candidate.

Which negative control is defensible?

Use Appendix as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 8-10% consistently across the quick facts, protocol table, and poster.

What transfer method to use for SMTN Western blot?

Use the transfer method in the recommended protocol and verify transfer before blocking.

How should A04895 be started?

Start at the linked datasheet condition and run a three-point primary-antibody dilution test.

Which publication-derived protocols can I compare?

Compare only the supplied target-verified records: PMC9025042, PMC12750176, PMC12886777.

Boster reagents

SMTN Western Blot Reagents

Human/Mouse/Rat-reactive SMTN Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for SMTN using A04895; observed band 99.1 kDa

Anti-Smoothelin/SMTN Antibody Picoband®

Cat # A04895

View product →

Real WB data Western blot validation image for SMTN using A04895-2; observed band 99.1 kDa

Anti-SMTN Antibody Picoband®

Cat # A04895-2