ID2 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 14.9 kDa observed
Observed band 14.9 kDa observed
Gel 15%
Positive control No defensible HPA tissue; use knockout/knockdown
Negative control Use knockout/knockdown or orthogonal negative
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass14.9 kDa
LocalizationCytoplasm / Nucleus
Processing / PTMRecord-dependent
ReactivityHuman / Rat / Mouse
Section 1

Real Curated ID2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateKnown-positive human lysate
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
PrimaryA00417-1 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 ID2 Western Blot Band Size?

Use the product-observed 14.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
14.9 kDaMatches the authoritative product WB observation.
14.9 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected ID2 appearancePlan around 14.9 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band14.9 kDa; use this as the primary experimental expectation.
Calculated mass14.9 kDa from UniProt Q02363; retain as context.
Gel selection15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00417-1.
Why is my band missing or off?
SituationLikely causeNext action
14.9 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 ID2 Western blot

🧪No defensible HPA positive control was available; do not invent one.
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for ID2

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

Undetected expression · recommended negative controls

The Human Protein Atlas exposes no structured IHC tissue profile for ID2, so tissue-level positive/negative controls cannot be listed here. Use a documented ID2-positive lysate (recombinant ID2 or a known-expressing line) as the positive control and a ID2 knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
Tissue Cell type Level Evidence Source
Section 3

Advanced ID2 Western Blot Tips

Deeper troubleshooting and optimisation questions for ID2, answered from its protein features.

Which band should guide the blot?
Use 14.9 kDa, the observation attached to the authoritative A00417-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 14.9 kDa expectation.
Which positive control should I start with?
No defensible positive tissue was supplied; do not invent one.
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 ID2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00417-1 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

ID2 Western Blot Reagents

Human/Rat/Mouse-reactive ID2 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for ID2 using A00417-1; observed band 14.9 kDa
Anti-ID2 Antibody Picoband®
Cat # A00417-1
Real WB dataWestern blot validation image for ID2 using A00417-3; observed band 14.9 kDa
Anti-ID2 Antibody Picoband®
Cat # A00417-3

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.