RPA1 · Western blot design guide

RPA1 Western Blot Planning Guide

Plan a RPA1 Western blot around the catalog-observed 68.1 kDa band, image-backed M01317-2 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 RPA1 (RPA1): expected band 68.1 kDa, antibody M01317-2, and PMC-cited SDS-PAGE protocol steps — RPA1 Western blot protocol sheet — expected band 68.1 kDa, antibody M01317-2, controls and PMC citations. Open the full RPA1 WB guide →

Navigate to: Protocols Band Controls Antibodies

RPA1 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	68.1 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Adipose tissue
	Negative control ⓘ	Target knockdown/knockout

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	68.1 kDa
ⓘ	Localization	Nucleus / Nucleus, PML body
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse

Section 1

Real Curated RPA1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Adipose tissue
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	M01317-2 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 RPA1 Western Blot Band Size?

Use the product-observed 68.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
68.1 kDa	Matches the authoritative product WB observation.
68.1 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 RPA1 appearancePlan around 68.1 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	68.1 kDa; use this as the primary experimental expectation.
Calculated mass	68.1 kDa from UniProt P27694; 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 M01317-2.

Why is my band missing or off?
Situation	Likely cause	Next action
68.1 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 P27694 Human Protein Atlas product datasheet (M01317-2)

Experiment evidence

Western blot images showing RPA1 band

M01317-2 · RPA1

← swipe / scroll for more images →

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

Sample controls for RPA1 Western blot

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

Positive control: Adipose tissue (High)

Negative control: Target knockdown/knockout

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

HPA tissue expression evidence for RPA1

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
Adipose tissue	Reported tissue cells	High	Protein (HPA)	HPA →
Breast	Reported tissue cells	High	Protein (HPA)	HPA →
Adrenal gland	Reported tissue cells	High	Protein (HPA)	HPA →
Bone marrow	Reported tissue cells	High	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source

Section 3

Advanced RPA1 Western Blot Tips

Deeper troubleshooting and optimisation questions for RPA1, answered from its protein features.

Which band should guide the blot?

Use 68.1 kDa, the observation attached to the authoritative M01317-2 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 68.1 kDa expectation.

Which positive control should I start with?

Start with Adipose tissue, 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 12-15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for RPA1 Western blot?

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

How should M01317-2 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: PMC12218172, PMC5986631, PMC13104110.

Boster reagents

RPA1 Western Blot Reagents

Human/Mouse-reactive RPA1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for RPA1 using M01317-2; observed band 68.1 kDa

Anti-RPA70 RPA1 Antibody Picoband® (monoclonal, 11H4)

Cat # M01317-2

View product →

Real WB data Western blot validation image for RPA1 using PB9886; observed band 68.1 kDa

Anti-RPA70/RPA1 Antibody Picoband®

Cat # PB9886