SUB1 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 14.4 kDa observed
Observed band 14.4 kDa observed
Gel 15%
Positive control ⓘ Breast
Negative control ⓘ Adipose tissue+4 more · see all
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass14.4 kDa
ⓘLocalizationNucleus
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Mouse / Rat
Section 1

Real Curated SUB1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAppendix
Gel %15%
Load20-30 µg total protein per lane
TransferSemi-dry, short transfer
Membrane0.2 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryM02698-1 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 SUB1 Western Blot Band Size?

Use the product-observed 14.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
14.4 kDaMatches the authoritative product WB observation.
14.4 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected SUB1 appearancePlan around 14.4 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band14.4 kDa; use this as the primary experimental expectation.
Calculated mass14.4 kDa from UniProt P53999; retain as context.
Gel selection15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with M02698-1.
Why is my band missing or off?
SituationLikely causeNext action
14.4 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 SUB1 Western blot

🧪Use Breast as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Breast (High)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for SUB1

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
Breast Reported tissue cells High Protein (HPA) HPA →
Cervix Reported tissue cells High Protein (HPA) HPA →
Cerebellum Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →
Skeletal muscle Reported tissue cells Low Protein (HPA) HPA →
Smooth muscle Reported tissue cells Low Protein (HPA) HPA →

Undetected expression · recommended negative controls

The available HPA profile lists no reliably-negative tissue for SUB1 (a broadly-expressed target). Use a SUB1 knockdown (siRNA/shRNA) or knockout lysate as the negative control and confirm signal loss versus the parental lysate.
Tissue Cell type Level Evidence Source
Section 3

Advanced SUB1 Western Blot Tips

Deeper troubleshooting and optimisation questions for SUB1, answered from its protein features.

Which band should guide the blot?
Use 14.4 kDa, the observation attached to the authoritative M02698-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 14.4 kDa expectation.
Which positive control should I start with?
Start with Breast, 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 SUB1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should M02698-1 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

SUB1 Western Blot Reagents

Human/Mouse/Rat-reactive SUB1 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for SUB1 using M02698-1; observed band 14.4 kDa
Anti-PC4/SUB1 Antibody Picoband® (monoclonal, 2D13E3)
Cat # M02698-1
Real WB dataWestern blot validation image for SUB1 using A02698-1; observed band 14.4 kDa
Anti-PC4/SUB1 Antibody Picoband®
Cat # A02698-1
Real WB dataWestern blot validation image for SUB1 using M02698-2; observed band 14.4 kDa
Anti-PC4/SUB1 Antibody Picoband® (monoclonal, 6B5B10)
Cat # M02698-2
Real WB dataWestern blot validation image for SUB1 using M02698-3; observed band 14.4 kDa
Anti-PC4/SUB1 Antibody Picoband® (monoclonal, 8D9D1)
Cat # M02698-3
Real WB dataWestern blot validation image for SUB1 using PB10098; observed band 14.4 kDa
Anti-PC4/SUB1 Antibody Picoband®
Cat # PB10098

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.