DDX21 · Western blot design guide

DDX21 Western Blot Planning Guide

Plan a DDX21 Western blot around the catalog-observed 61.6 kDa band, image-backed A04605-2 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for DDX21 (DDX21): expected band 61.6 kDa, antibody A04605-2, and guide-derived SDS-PAGE protocol steps
DDX21 Western blot protocol sheet — expected band 61.6 kDa, antibody A04605-2, controls and PMC citations. Open the full DDX21 WB guide →

DDX21 Western Blot Experimental Design Guide

Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running
Expected band 61.6 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Colon
Negative control ⓘ Soft tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 61.6 kDa
ⓘ Localization Nucleus, nucleolus
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human
Section 1

Real Curated DDX21 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAppendix
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
PrimaryA04605-2 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 DDX21 Western Blot Band Size?

Use the product-observed 61.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
61.6 kDaMatches the authoritative product WB observation.
61.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected DDX21 appearancePlan around 61.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band61.6 kDa; use this as the primary experimental expectation.
Calculated mass61.6 kDa from UniProt Q9NY93; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A04605-2.
Why is my band missing or off?
SituationLikely causeNext action
61.6 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 DDX21 Western blot

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

HPA tissue expression evidence for DDX21

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Colon Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells High Protein (HPA) HPA →
Duodenum Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Soft tissue Reported tissue cells Not detected Protein (HPA) HPA →
Ovary Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced DDX21 Western Blot Tips

Deeper troubleshooting and optimisation questions for DDX21, answered from its protein features.

Which band should guide the blot?
Use 61.6 kDa, the observation attached to the authoritative A04605-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 61.6 kDa expectation.
Which positive control should I start with?
Start with Colon, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Soft tissue 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 DDX21 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04605-2 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

DDX21 Western Blot Reagents

Human-reactive DDX21 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for DDX21 using A04605-2; observed band 61.6 kDa
Anti-DDX21 Antibody Picoband®
Cat # A04605-2

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.

References

  1. UniProt Q9NY93
  2. Human Protein Atlas — DDX21
  3. A04605-2 product record
  4. PMC13284300 — Ubiquitination of DDX21 by HERC2 induces a dormancy-like phenotype via the NUCKS1-p21/p27 axis to promote radio-resistance in colorectal cancer cells (Cell death & disease, 2026)
  5. PMC6754283 — Activation of PARP-1 by snoRNAs Controls Ribosome Biogenesis and Cell Growth via the RNA Helicase DDX21 (Molecular cell, 2019)
  6. PMC10877017 — DEAD-box RNA helicase 21 interacts with porcine circovirus type 2 Cap protein and facilitates viral replication (Frontiers in microbiology, 2024)