IGFBP3 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 31.7 kDa observed
Observed band 31.7 kDa observed
Gel 12-15%
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 mass31.7 kDa
LocalizationSecreted / Nucleus
Processing / PTMRecord-dependent
ReactivityHuman / Rat / Mouse
Section 1

Real Curated IGFBP3 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 %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
PrimaryA00435-2 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 IGFBP3 Western Blot Band Size?

Use the product-observed 31.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
31.7 kDaMatches the authoritative product WB observation.
31.7 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected IGFBP3 appearancePlan around 31.7 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band31.7 kDa; use this as the primary experimental expectation.
Calculated mass31.7 kDa from UniProt P17936; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00435-2.
Why is my band missing or off?
SituationLikely causeNext action
31.7 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 IGFBP3 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 IGFBP3

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 IGFBP3, so tissue-level positive/negative controls cannot be listed here. Use a documented IGFBP3-positive lysate (recombinant IGFBP3 or a known-expressing line) as the positive control and a IGFBP3 knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
Tissue Cell type Level Evidence Source
Section 3

Advanced IGFBP3 Western Blot Tips

Deeper troubleshooting and optimisation questions for IGFBP3, answered from its protein features.

Which band should guide the blot?
Use 31.7 kDa, the observation attached to the authoritative A00435-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 31.7 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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for IGFBP3 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00435-2 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

IGFBP3 Western Blot Reagents

Human/Rat/Mouse-reactive IGFBP3 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for IGFBP3 using A00435-2; observed band 31.7 kDa
Anti-IGFBP3 Antibody Picoband®
Cat # A00435-2
Real WB dataWestern blot validation image for IGFBP3 using PA1498; observed band 31.7 kDa
Anti-IGFBP3 Antibody Picoband®
Cat # PA1498

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.