DDAH2 · Western blot design guide

DDAH2 Western Blot Planning Guide

Plan a DDAH2 Western blot around the catalog-observed 29.6 kDa band, image-backed PB10001 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 DDAH2 (DDAH2): expected band 29.6 kDa, antibody PB10001, and PMC-cited SDS-PAGE protocol steps

Band Protocols Controls Antibodies

DDAH2 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	29.6 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Breast
	Negative control ⓘ	Soft tissue

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	29.6 kDa
ⓘ	Localization	Cytoplasm / Mitochondrion
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Sample controls for DDAH2 Western blot

🧪Use Breast as the first positive-control candidate and Soft tissue as the HPA Not detected negative candidate.

Positive control: Breast (Medium)

Negative control: Soft tissue (Not detected)

HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for DDAH2

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
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	Medium	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Soft tissue	Reported tissue cells	Not detected	Protein (HPA)	HPA →

Section 1

What Is the Expected DDAH2 Western Blot Band Size?

Use the product-observed 29.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
29.6 kDa	Matches the authoritative product WB observation.
29.6 kDa calculated	Use as UniProt context, not as a replacement observed band.
Unexpected additional signal	Do not assign identity without orthogonal positive/negative controls.

💡Expected DDAH2 appearancePlan around 29.6 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	29.6 kDa; use this as the primary experimental expectation.
Calculated mass	29.6 kDa from UniProt O95865; retain as context.
Gel selection	12-15%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with PB10001.

Why is my band missing or off?
Situation	Likely cause	Next action
29.6 kDa	Matches the authoritative product WB observation.	Confirm with orthogonal controls and the linked product record.
Additional band	May reflect processing, modification, or non-specific signal.	Run a dilution series and compare positive/negative controls.
Weak signal	Target abundance or transfer may be limiting.	Verify transfer, increase positive-control abundance, and bracket exposure.

Sources: UniProt O95865 Human Protein Atlas product datasheet (PB10001)

Experiment evidence

Western blot images showing DDAH2 band

PB10001 · DDAH2 — observed 29.6 kDa WB validation

PB10001 · DDAH2Observed 29.6 kDa · product-specific WB image

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Section 2

Real Curated DDAH2 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Adrenal gland
Gel %	12-15%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, short transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	PB10001 at datasheet starting dilution
Primary incubation	Overnight at 4 °C with gentle agitation
Secondary	Species-matched HRP conjugate at validated dilution
Wash	3 × 5 min in TBST
Detection	Chemiluminescent substrate
Exposure	Bracket exposures to avoid saturation

Section 4

Advanced DDAH2 Western Blot Tips

Deeper troubleshooting and optimisation questions for DDAH2, answered from its protein features.

Which band should guide the blot?

Use 29.6 kDa, the observation attached to the authoritative PB10001 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 29.6 kDa expectation.

Which positive control should I start with?

Start with Breast, 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 DDAH2 Western blot?

Use the transfer method in the recommended protocol and verify transfer before blocking.

How should PB10001 be started?

Start at the linked datasheet condition and run a three-point primary-antibody dilution test.

Which publication-derived protocols can I compare?

Compare only the supplied target-verified records: PMC5427783, PMC3818215, PMC5245815.

Boster reagents

DDAH2 Western Blot Reagents

Human/Mouse/Rat-reactive DDAH2 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for DDAH2 using PB10001; observed band 29.6 kDa

Anti-DDAH2 Antibody Picoband®

Cat # PB10001

Reactivity: Human, Mouse, Rat

View product →

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.