MT-ND4 · Western blot design guide

MT-ND4 Western Blot Planning Guide

Plan a MT-ND4 Western blot around the catalog-observed 51.6 kDa band, image-backed A04180-1 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 MT-ND4 (MT-ND4): expected band 51.6 kDa, antibody A04180-1, and PMC-cited SDS-PAGE protocol steps — MT-ND4 Western blot protocol sheet — expected band 51.6 kDa, antibody A04180-1, controls and PMC citations. Open the full MT-ND4 WB guide →

Navigate to: Protocols Band Controls Antibodies

MT-ND4 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	51.6 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Kidney
	Negative control ⓘ	Bone marrow

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	51.6 kDa
ⓘ	Localization	Mitochondrion inner membrane
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human

Section 1

Real Curated MT-ND4 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysate	Duodenum
Gel %	12-15%
Load	20-30 µg total protein per lane
Transfer	Semi-dry, standard transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A04180-1 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 2

What Is the Expected MT-ND4 Western Blot Band Size?

Use the product-observed 51.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
51.6 kDa	Matches the authoritative product WB observation.
51.6 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 MT-ND4 appearancePlan around 51.6 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	51.6 kDa; use this as the primary experimental expectation.
Calculated mass	51.6 kDa from UniProt P03905; 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 A04180-1.

Why is my band missing or off?
Situation	Likely cause	Next action
51.6 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 P03905 Human Protein Atlas product datasheet (A04180-1)

Experiment evidence

Western blot images showing MT-ND4 band

A04180-1 · MT-ND4

← swipe / scroll for more images →

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

Sample controls for MT-ND4 Western blot

🧪Use Kidney as the first positive-control candidate and Bone marrow as the HPA Not detected negative candidate.

Positive control: Kidney (High)

Negative control: Bone marrow (Not detected)

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

HPA tissue expression evidence for MT-ND4

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

Tissue	Cell type	Level	Evidence	Source
Kidney	Reported tissue cells	High	Protein (HPA)	HPA →
Stomach	Reported tissue cells	High	Protein (HPA)	HPA →
Small intestine	Reported tissue cells	High	Protein (HPA)	HPA →
Parathyroid gland	Reported tissue cells	High	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

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

Section 3

Advanced MT-ND4 Western Blot Tips

Deeper troubleshooting and optimisation questions for MT-ND4, answered from its protein features.

Which band should guide the blot?

Use 51.6 kDa, the observation attached to the authoritative A04180-1 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 51.6 kDa expectation.

Which positive control should I start with?

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

Which negative control is defensible?

Use Bone marrow 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 MT-ND4 Western blot?

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

How should A04180-1 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: PMC13216193, PMC9326446, PMC12559409.

Boster reagents

MT-ND4 Western Blot Reagents

Human-reactive MT-ND4 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for MT-ND4 using A04180-1; observed band 51.6 kDa

Anti-MT-ND4 Antibody Picoband®

Cat # A04180-1

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.