RPS6KB1 · Western blot design guide

RPS6KB1 Western Blot Planning Guide

Plan a RPS6KB1 Western blot around the catalog-observed 59.1 kDa band, image-backed A01475-2 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 RPS6KB1 (RPS6KB1): expected band 59.1 kDa, antibody A01475-2, and PMC-cited SDS-PAGE protocol steps — RPS6KB1 Western blot protocol sheet — expected band 59.1 kDa, antibody A01475-2, controls and PMC citations. Open the full RPS6KB1 WB guide →

Navigate to: Protocols Band Controls Antibodies

RPS6KB1 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	59.1 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Cerebellum
	Negative control ⓘ	Soft tissue

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	59.1 kDa
ⓘ	Localization	Synapse, synaptosome / Mitochondrion outer membrane
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse

Section 1

Real Curated RPS6KB1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Breast
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	A01475-2 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 RPS6KB1 Western Blot Band Size?

Use the product-observed 59.1 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
59.1 kDa	Matches the authoritative product WB observation.
59.1 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 RPS6KB1 appearancePlan around 59.1 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	59.1 kDa; use this as the primary experimental expectation.
Calculated mass	59.1 kDa from UniProt P23443; 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 A01475-2.

Why is my band missing or off?
Situation	Likely cause	Next action
59.1 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 P23443 Human Protein Atlas product datasheet (A01475-2)

Experiment evidence

Western blot images showing RPS6KB1 band

A01475-2 · RPS6KB1

← swipe / scroll for more images →

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

Sample controls for RPS6KB1 Western blot

🧪Use Cerebellum as the first positive-control candidate and Soft tissue as the HPA Not detected negative candidate.

Positive control: Cerebellum (High)

Negative control: Soft tissue (Not detected)

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

HPA tissue expression evidence for RPS6KB1

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
Cerebellum	Reported tissue cells	High	Protein (HPA)	HPA →
Breast	Reported tissue cells	High	Protein (HPA)	HPA →
Bronchus	Reported tissue cells	High	Protein (HPA)	HPA →
Cerebral cortex	Reported tissue cells	High	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

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

Section 3

Advanced RPS6KB1 Western Blot Tips

Deeper troubleshooting and optimisation questions for RPS6KB1, answered from its protein features.

Which band should guide the blot?

Use 59.1 kDa, the observation attached to the authoritative A01475-2 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 59.1 kDa expectation.

Which positive control should I start with?

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

Which negative control is defensible?

Use Soft tissue 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 RPS6KB1 Western blot?

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

How should A01475-2 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: PMC5549961, PMC7371735, PMC5050127.

Boster reagents

RPS6KB1 Western Blot Reagents

Human/Mouse-reactive RPS6KB1 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for RPS6KB1 using A01475-2; observed band 59.1 kDa

Anti-S6K1/RPS6KB1 Antibody Picoband®

Cat # A01475-2

View product →

Real WB data Western blot validation image for RPS6KB1 using A01475-3; observed band 59.1 kDa

Anti-p70 S6K/RPS6KB1 Antibody Picoband®

Cat # A01475-3