PRKAA1 · Western blot design guide

PRKAA1 Western Blot Planning Guide

Plan a PRKAA1 Western blot around the catalog-observed 64 kDa band, image-backed A00994-3 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 PRKAA1 (PRKAA1): expected band 64 kDa, antibody A00994-3, and PMC-cited SDS-PAGE protocol steps — PRKAA1 Western blot protocol sheet — expected band 64 kDa, antibody A00994-3, controls and PMC citations. Open the full PRKAA1 WB guide →

Navigate to: Protocols Band Controls Antibodies

PRKAA1 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	64 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Colon
	Negative control ⓘ	Bone marrow

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	64 kDa
ⓘ	Localization	Cytoplasm / Nucleus
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Section 1

Real Curated PRKAA1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Adrenal gland
Gel %	12-15%
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	A00994-3 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 PRKAA1 Western Blot Band Size?

Use the product-observed 64 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
64 kDa	Matches the authoritative product WB observation.
64 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 PRKAA1 appearancePlan around 64 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	64 kDa; use this as the primary experimental expectation.
Calculated mass	64 kDa from UniProt Q13131; retain as context.
Gel selection	12-15%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A00994-3.

Why is my band missing or off?
Situation	Likely cause	Next action
64 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 Q13131 Human Protein Atlas product datasheet (A00994-3)

Experiment evidence

Western blot images showing PRKAA1 band

A00994-3 · PRKAA1

← swipe / scroll for more images →

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

Sample controls for PRKAA1 Western blot

🧪Use Colon as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.

Positive control: Colon (Medium)

Negative control: Bone marrow (Not detected)

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

HPA tissue expression evidence for PRKAA1

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
Colon	Reported tissue cells	Medium	Protein (HPA)	HPA →
Adrenal gland	Reported tissue cells	Medium	Protein (HPA)	HPA →
Appendix	Reported tissue cells	Medium	Protein (HPA)	HPA →
Bronchus	Reported tissue cells	Medium	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

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

Section 3

Advanced PRKAA1 Western Blot Tips

Deeper troubleshooting and optimisation questions for PRKAA1, answered from its protein features.

Which band should guide the blot?

Use 64 kDa, the observation attached to the authoritative A00994-3 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 64 kDa expectation.

Which positive control should I start with?

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

Which negative control is defensible?

Use Bone marrow as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 12-15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for PRKAA1 Western blot?

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

How should A00994-3 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: PMC5559581, PMC8726653, PMC9059102.

Boster reagents

PRKAA1 Western Blot Reagents

Human/Mouse/Rat-reactive PRKAA1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for PRKAA1 using A00994-3; observed band 64 kDa

Anti-AMPK alpha 1/PRKAA1 Antibody Picoband®

Cat # A00994-3

View product →

Real WB data Western blot validation image for PRKAA1 using A00994-6; observed band 64 kDa