PFN1 · Western blot design guide

PFN1 Western Blot Planning Guide

Plan a PFN1 Western blot around the catalog-observed 15.1 kDa band, image-backed PB9313 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 PFN1 (PFN1): expected band 15.1 kDa, antibody PB9313, and PMC-cited SDS-PAGE protocol steps — PFN1 Western blot protocol sheet — expected band 15.1 kDa, antibody PB9313, controls and PMC citations. Open the full PFN1 WB guide →

Navigate to: Protocols Band Controls Antibodies

PFN1 Western Blot Experimental Design Guide

Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running

	Expected band	15.1 kDa
	Observed band	Not reported — verify product WB image
	Gel	15%
	Positive control ⓘ	Spleen
	Negative control ⓘ	Target knockdown/knockout

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	15.1 kDa
ⓘ	Localization	Cytoplasm, cytoskeleton
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Section 1

Real Curated PFN1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Lung
Gel %	15%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, short transfer
Membrane	0.2 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	PB9313 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 PFN1 Western Blot Band Size?

Use the product-observed 15.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
15.1 kDa	Matches the authoritative product WB observation.
15.1 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 PFN1 appearancePlan around 15.1 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	15.1 kDa; use this as the primary experimental expectation.
Calculated mass	15.1 kDa from UniProt P07737; retain as context.
Gel selection	15%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with PB9313.

Why is my band missing or off?
Situation	Likely cause	Next action
15.1 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 P07737 Human Protein Atlas product datasheet (PB9313)

Experiment evidence

Western blot images showing PFN1 band

PB9313 · PFN1

← swipe / scroll for more images →

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

Sample controls for PFN1 Western blot

🧪Use Spleen as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.

Positive control: Spleen (High)

Negative control: Target knockdown/knockout

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

HPA tissue expression evidence for PFN1

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
Spleen	Reported tissue cells	High	Protein (HPA)	HPA →
Lung	Reported tissue cells	High	Protein (HPA)	HPA →
Lymph node	Reported tissue cells	High	Protein (HPA)	HPA →
Tonsil	Reported tissue cells	High	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source

Section 3

Advanced PFN1 Western Blot Tips

Deeper troubleshooting and optimisation questions for PFN1, answered from its protein features.

Which band should guide the blot?

Use 15.1 kDa, the observation attached to the authoritative PB9313 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 15.1 kDa expectation.

Which positive control should I start with?

Start with Spleen, the lead HPA protein-expression candidate.

Which negative control is defensible?

Use a target knockdown/knockout control when no HPA Not detected tissue is available.

Which gel should I use?

Use 15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for PFN1 Western blot?

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

How should PB9313 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: PMC6125445, PMC2783708.

Boster reagents

PFN1 Western Blot Reagents

Human/Mouse/Rat-reactive PFN1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for PFN1 using PB9313; observed band 15.1 kDa

Anti-Profilin 1/PFN1 Antibody Picoband®

Cat # PB9313

View product →

Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.

Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.