CD44 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 81.5 kDa observed
Observed band 81.5 kDa observed
Gel 8-10%
Positive control Cervix
Negative control Endometrium
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass81.5 kDa
LocalizationCell membrane / Cell projection, microvillus
Processing / PTMRecord-dependent
ReactivityHuman / Mouse / Rat
Section 1

Real Curated CD44 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAppendix
Gel %8-10%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA00052 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 CD44 Western Blot Band Size?

Use the product-observed 81.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
81.5 kDaMatches the authoritative product WB observation.
81.5 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected CD44 appearancePlan around 81.5 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band81.5 kDa; use this as the primary experimental expectation.
Calculated mass81.5 kDa from UniProt P16070; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00052.
Why is my band missing or off?
SituationLikely causeNext action
81.5 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 CD44 Western blot

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

HPA tissue expression evidence for CD44

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
Cervix Reported tissue cells High Protein (HPA) HPA →
Breast Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced CD44 Western Blot Tips

Deeper troubleshooting and optimisation questions for CD44, answered from its protein features.

Which band should guide the blot?
Use 81.5 kDa, the observation attached to the authoritative A00052 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 81.5 kDa expectation.
Which positive control should I start with?
Start with Cervix, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Endometrium as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for CD44 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00052 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

CD44 Western Blot Reagents

Human/Mouse/Rat-reactive CD44 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for CD44 using A00052; observed band 81.5 kDa
Anti-CD44 Antibody Picoband®
Cat # A00052
Real WB dataWestern blot validation image for CD44 using A00052-2; observed band 81.5 kDa
Anti-CD44 Antibody Picoband®
Cat # A00052-2
Real WB dataWestern blot validation image for CD44 using M00052-3; observed band 81.5 kDa
Anti-CD44 Picoband® Antibody (monoclonal, 7H7)
Cat # M00052-3
Real WB dataWestern blot validation image for CD44 using PA1021-2; observed band 81.5 kDa
Anti-CD44 antigen CD44 Antibody Picoband®
Cat # PA1021-2
Real WB dataWestern blot validation image for CD44 using PB9333; observed band 81.5 kDa
Anti-CD44 Antibody Picoband®
Cat # PB9333

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.