APEX1 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 35.6 kDa observed
Observed band 35.6 kDa observed
Gel 12-15%
Positive control Adrenal gland
Negative control Adipose tissue+4 more · see all
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass35.6 kDa
LocalizationNucleus / Nucleus, nucleolus
Processing / PTMRecord-dependent
ReactivityHuman / Rat / Mouse
Section 1

Real Curated APEX1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
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
PrimaryM00627 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 APEX1 Western Blot Band Size?

Use the product-observed 35.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
35.6 kDaMatches the authoritative product WB observation.
35.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected APEX1 appearancePlan around 35.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band35.6 kDa; use this as the primary experimental expectation.
Calculated mass35.6 kDa from UniProt P27695; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with M00627.
Why is my band missing or off?
SituationLikely causeNext action
35.6 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 APEX1 Western blot

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

HPA tissue expression evidence for APEX1

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
Adrenal gland Reported tissue cells High Protein (HPA) HPA →
Breast Reported tissue cells High Protein (HPA) HPA →
Colon Reported tissue cells High Protein (HPA) HPA →
Cerebellum Reported tissue cells High Protein (HPA) HPA →
Heart 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 APEX1 (a broadly-expressed target). Use a APEX1 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 APEX1 Western Blot Tips

Deeper troubleshooting and optimisation questions for APEX1, answered from its protein features.

Which band should guide the blot?
Use 35.6 kDa, the observation attached to the authoritative M00627 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 35.6 kDa expectation.
Which positive control should I start with?
Start with Adrenal gland, 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 APEX1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should M00627 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

APEX1 Western Blot Reagents

Human/Rat/Mouse-reactive APEX1 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for APEX1 using M00627; observed band 35.6 kDa
Anti-APE1 APEX1 Antibody Picoband® (monoclonal, 5C11)
Cat # M00627
Real WB dataWestern blot validation image for APEX1 using PA1494; observed band 35.6 kDa
Anti-APE1/APEX1 Antibody Picoband®
Cat # PA1494
Real WB dataWestern blot validation image for APEX1 using PB9128; observed band 35.6 kDa
Anti-APE1/APEX1 Antibody Picoband®
Cat # PB9128

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.