DPH5 · Western blot design guide

DPH5 Western Blot Planning Guide

Plan a DPH5 Western blot around the catalog-observed 31.7 kDa band, image-backed A11989-1 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 DPH5 (DPH5): expected band 31.7 kDa, antibody A11989-1, and guide-derived SDS-PAGE protocol steps
DPH5 Western blot protocol sheet — expected band 31.7 kDa, antibody A11989-1, controls and PMC citations. Open the full DPH5 WB guide →

DPH5 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 band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Adrenal gland
Negative control ⓘ Lymph node
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 31.7 kDa
ⓘ Localization Not reported
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human
Section 1

Real Curated DPH5 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
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
PrimaryA11989-1 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 DPH5 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 DPH5 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 Q9H2P9; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A11989-1.
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 DPH5 Western blot

🧪Use Adrenal gland as the first positive-control candidate and Lymph node as the HPA Not detected negative candidate.
Positive control: Adrenal gland (Medium)
Negative control: Lymph node (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for DPH5

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
Adrenal gland Reported tissue cells Medium Protein (HPA) HPA →
Cerebellum Reported tissue cells Medium Protein (HPA) HPA →
Caudate Reported tissue cells Medium Protein (HPA) HPA →
Appendix Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

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

Advanced DPH5 Western Blot Tips

Deeper troubleshooting and optimisation questions for DPH5, answered from its protein features.

Which band should guide the blot?
Use 31.7 kDa, the observation attached to the authoritative A11989-1 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?
Start with Adrenal gland, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Lymph node 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 DPH5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A11989-1 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

DPH5 Western Blot Reagents

Human-reactive DPH5 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for DPH5 using A11989-1; observed band 31.7 kDa
Anti-DPH5 Antibody Picoband®
Cat # A11989-1

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 Q9H2P9
  2. Human Protein Atlas — DPH5
  3. A11989-1 product record
  4. PMC5371833 — Influence of DPH1 and DPH5 Protein Variants on the Synthesis of Diphthamide, the Target of ADPRibosylating Toxins (Toxins, 2017)
  5. PMC11955877 — Comparative Analysis of Two NNMT Bisubstrate Inhibitors through Chemoproteomic Studies: Uncovering the Role of Unconventional SAM Analogue Moiety for Improved Selectivity (ACS chemical biology, 2024)