BCKDHA · Western blot design guide

BCKDHA Western Blot Planning Guide

Plan a BCKDHA Western blot around the catalog-observed 50.5 kDa band, image-backed A04561-1 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 BCKDHA (BCKDHA): expected band 50.5 kDa, antibody A04561-1, and PMC-cited SDS-PAGE protocol steps

Band Protocols Controls Antibodies

BCKDHA 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	50.5 kDa
	Observed band	Not reported — verify product WB image
	Gel	12-15%
	Positive control ⓘ	Adrenal gland
	Negative control ⓘ	Target knockdown/knockout

Important caveats

Reasons your observed band may differ from the expected size.

ⓘ	Calculated mass	50.5 kDa
ⓘ	Localization	Mitochondrion matrix
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Sample controls for BCKDHA Western blot

🧪Use Adrenal gland as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.

Positive control: Adrenal gland (High)

Negative control: Target knockdown/knockout

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

HPA tissue expression evidence for BCKDHA

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
Adrenal gland	Reported tissue cells	High	Protein (HPA)	HPA →
Breast	Reported tissue cells	High	Protein (HPA)	HPA →
Appendix	Reported tissue cells	High	Protein (HPA)	HPA →
Bone marrow	Reported tissue cells	High	Protein (HPA)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source

No HPA Not detected tissue is available; use a target knockdown/knockout negative control.

Section 1

What Is the Expected BCKDHA Western Blot Band Size?

Use the product-observed 50.5 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
50.5 kDa	Matches the authoritative product WB observation.
50.5 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 BCKDHA appearancePlan around 50.5 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	50.5 kDa; use this as the primary experimental expectation.
Calculated mass	50.5 kDa from UniProt P12694; 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 A04561-1.

Why is my band missing or off?
Situation	Likely cause	Next action
50.5 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 P12694 Human Protein Atlas product datasheet (A04561-1)

Experiment evidence

Western blot images showing BCKDHA band

A04561-1 · BCKDHA — observed 50.5 kDa WB validation

A04561-1 · BCKDHAObserved 50.5 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 BCKDHA 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, standard transfer
Membrane	0.45 µm PVDF
Blocking	5% non-fat milk or 5% BSA in TBST
Primary	A04561-1 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 BCKDHA Western Blot Tips

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

Which band should guide the blot?

Use 50.5 kDa, the observation attached to the authoritative A04561-1 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 50.5 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 a target knockdown/knockout control when no HPA Not detected tissue is available.

Which gel should I use?

Use 12-15% consistently across the quick facts, protocol table, and poster.

What transfer method to use for BCKDHA Western blot?

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

How should A04561-1 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: PMC13042768, PMC9174284, PMC11194096.

Boster reagents

BCKDHA Western Blot Reagents

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

Real WB data Western blot validation image for BCKDHA using A04561-1; observed band 50.5 kDa

Anti-BCKDHA Antibody Picoband®

Cat # A04561-1

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.