EVA1A · Western blot design guide

EVA1A Western Blot Planning Guide

Plan an EVA1A Western blot around the catalog-observed 17.5 kDa band, image-backed A11580-1 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 EVA1A (EVA1A): expected band 17.5 kDa, antibody A11580-1, and PMC-cited SDS-PAGE protocol steps

Band Protocols Controls Antibodies

EVA1A 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	17.5 kDa
	Observed band	Not reported — verify product WB image
	Gel	15%
	Positive control ⓘ	Liver
	Negative control ⓘ	Appendix

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	17.5 kDa
ⓘ	Localization	Endoplasmic reticulum membrane / Lysosome membrane
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Sample controls for EVA1A Western blot

🧪Use Liver as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.

Positive control: Liver (High)

Negative control: Appendix (Not detected)

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

HPA tissue expression evidence for EVA1A

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
Liver	Reported tissue cells	High	Protein (HPA)	HPA →
Testis	Reported tissue cells	High	Protein (HPA)	HPA →
Kidney	Reported tissue cells	Medium	Protein (HPA)	HPA →
Adrenal gland	Reported tissue cells	Low	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

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

Section 1

What Is the Expected EVA1A Western Blot Band Size?

Use the product-observed 17.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
17.5 kDa	Matches the authoritative product WB observation.
17.5 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 EVA1A appearancePlan around 17.5 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	17.5 kDa; use this as the primary experimental expectation.
Calculated mass	17.5 kDa from UniProt Q9H8M9; retain as context.
Gel selection	15%; shared with the recommended protocol and poster.
Specificity check	Compare the lead HPA positive and negative controls with A11580-1.

Why is my band missing or off?
Situation	Likely cause	Next action
17.5 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 Q9H8M9 Human Protein Atlas product datasheet (A11580-1)

Experiment evidence

Western blot images showing EVA1A band

A11580-1 · EVA1A — observed 17.5 kDa WB validation

A11580-1 · EVA1AObserved 17.5 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 EVA1A Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Liver
Gel %	15%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, short transfer
Membrane	0.2 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A11580-1 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 EVA1A Western Blot Tips

Deeper troubleshooting and optimisation questions for EVA1A, answered from its protein features.

Which band should guide the blot?

Use 17.5 kDa, the observation attached to the authoritative A11580-1 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 17.5 kDa expectation.

Which positive control should I start with?

Start with Liver, the lead HPA protein-expression candidate.

Which negative control is defensible?

Use Appendix as an orthogonal HPA Not detected candidate.

Which gel should I use?

Use 15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for EVA1A Western blot?

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

How should A11580-1 be started?

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

Which publication-derived protocols can I compare?

Compare only the supplied target-verified records: PMC12645495, PMC6039435, PMC5108317.

Boster reagents

EVA1A Western Blot Reagents

Human/Mouse/Rat-reactive EVA1A Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for EVA1A using A11580-1; observed band 17.5 kDa

Anti-TMEM166/EVA1A Antibody Picoband®

Cat # A11580-1

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.