LIF 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 22 kDa observed
Observed band 22 kDa observed
Gel 12-15%
Positive control Adipose tissue
Negative control Smooth muscle
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass22 kDa
LocalizationSecreted
Processing / PTMRecord-dependent
ReactivityMouse / Rat / Human
Section 1

Real Curated LIF Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdipose tissue
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, short transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryPA1562 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 LIF Western Blot Band Size?

Use the product-observed 22 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
22 kDaMatches the authoritative product WB observation.
22 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected LIF appearancePlan around 22 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band22 kDa; use this as the primary experimental expectation.
Calculated mass22 kDa from UniProt P15018; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with PA1562.
Why is my band missing or off?
SituationLikely causeNext action
22 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 LIF Western blot

🧪Use Adipose tissue as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Adipose tissue (Medium)
Negative control: Smooth muscle (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for LIF

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
Adipose tissue Reported tissue cells Medium Protein (HPA) HPA →
Breast Reported tissue cells Medium Protein (HPA) HPA →
Adrenal gland Reported tissue cells Medium Protein (HPA) HPA →
Appendix Reported tissue cells Medium Protein (HPA) HPA →
Bone marrow Reported tissue cells Low Protein (HPA) HPA →

Undetected expression · recommended negative controls

Tissue Cell type Level Evidence Source
Smooth muscle Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced LIF Western Blot Tips

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

Which band should guide the blot?
Use 22 kDa, the observation attached to the authoritative PA1562 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 22 kDa expectation.
Which positive control should I start with?
Start with Adipose tissue, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Smooth muscle 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 LIF Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PA1562 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

LIF Western Blot Reagents

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

Real WB dataWestern blot validation image for LIF using PA1562; observed band 22 kDa
Anti-Leukemia inhibitory factor LIF Antibody Picoband®
Cat # PA1562
Real WB dataWestern blot validation image for LIF using PB9036; observed band 22 kDa
Anti-LIF Antibody Picoband®
Cat # PB9036

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.