RNF8 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 55.5 kDa observed
Observed band 55.5 kDa observed
Gel 12-15%
Positive control ⓘ Cerebellum
Negative control ⓘ Bone marrow
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass55.5 kDa
ⓘLocalizationNucleus / Cytoplasm
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Mouse
Section 1

Real Curated RNF8 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateCerebellum
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA00707-2 at datasheet starting dilution
Primary incubationOvernight at 4 °C with gentle agitation
SecondarySpecies-matched HRP conjugate at validated dilution
Wash3 × 5 min in TBST
DetectionChemiluminescent substrate
ExposureBracket exposures to avoid saturation
Section 2

What Is the Expected RNF8 Western Blot Band Size?

Use the product-observed 55.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
55.5 kDaMatches the authoritative product WB observation.
55.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected RNF8 appearancePlan around 55.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band55.5 kDa; use this as the primary experimental expectation.
Calculated mass55.5 kDa from UniProt O76064; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00707-2.
Why is my band missing or off?
SituationLikely causeNext action
55.5 kDaMatches the authoritative product WB observation.Confirm with orthogonal controls and the linked product record.
Additional bandMay reflect processing, modification, or non-specific signal.Run a dilution series and compare positive/negative controls.
Weak signalTarget abundance or transfer may be limiting.Verify transfer, increase positive-control abundance, and bracket exposure.

Sample controls for RNF8 Western blot

🧪Use Cerebellum as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Cerebellum (High)
Negative control: Bone marrow (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for RNF8

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart on each tissue page — 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 →
Skin Reported tissue cells High Protein (HPA) HPA →
Kidney Reported tissue cells High Protein (HPA) HPA →
Stomach Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Lymph node Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced RNF8 Western Blot Tips

Deeper troubleshooting and optimisation questions for RNF8, answered from its protein features.

Which band should guide the blot?
Use 55.5 kDa, the observation attached to the authoritative A00707-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 55.5 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, 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 RNF8 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00707-2 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents

RNF8 Western Blot Reagents

Human/Mouse-reactive RNF8 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for RNF8 using A00707-2; observed band 55.5 kDa
Anti-RNF8 Antibody Picoband®
Cat # A00707-2
Real WB dataWestern blot validation image for RNF8 using A00707-3; observed band 55.5 kDa
Anti-RNF8 Antibody Picoband®
Cat # A00707-3

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.