KERA 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 40.5 kDa observed
Observed band 40.5 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 mass40.5 kDa
ⓘLocalizationSecreted, extracellular space, extracellular matrix
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Mouse
Section 1

Real Curated KERA 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
PrimaryPB9653 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 KERA Western Blot Band Size?

Use the product-observed 40.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
40.5 kDaMatches the authoritative product WB observation.
40.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected KERA appearancePlan around 40.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band40.5 kDa; use this as the primary experimental expectation.
Calculated mass40.5 kDa from UniProt O60938; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with PB9653.
Why is my band missing or off?
SituationLikely causeNext action
40.5 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 KERA 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 KERA

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

Advanced KERA Western Blot Tips

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

Which band should guide the blot?
Use 40.5 kDa, the observation attached to the authoritative PB9653 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 40.5 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 KERA Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9653 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

KERA Western Blot Reagents

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

Real WB dataWestern blot validation image for KERA using PB9653; observed band 40.5 kDa
Anti-Keratocan Antibody Picoband®
Cat # PB9653

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.