BCAT2 · Western blot design guide

BCAT2 Western Blot Planning Guide

Plan a BCAT2 Western blot around the catalog-observed 44.3 kDa band, image-backed A06369-1 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 BCAT2 (BCAT2): expected band 44.3 kDa observed, antibody A06369-1, and PMC-cited SDS-PAGE protocol steps
BCAT2 Western blot protocol sheet — expected band 44.3 kDa observed, antibody A06369-1, controls and PMC citations. Open the full BCAT2 WB guide →

BCAT2 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 44.3 kDa observed
Observed band 44.3 kDa observed
Gel 12-15%
Positive control ⓘ Adrenal gland
Negative control ⓘ Bone marrow
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 44.3 kDa
ⓘ Localization Mitochondrion
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated BCAT2 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
PrimaryA06369-1 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 BCAT2 Western Blot Band Size?

Use the product-observed 44.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
44.3 kDaMatches the authoritative product WB observation.
44.3 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected BCAT2 appearancePlan around 44.3 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band44.3 kDa; use this as the primary experimental expectation.
Calculated mass44.3 kDa from UniProt O15382; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A06369-1.
Why is my band missing or off?
SituationLikely causeNext action
44.3 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 BCAT2 Western blot

🧪Use Adrenal gland as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Adrenal gland (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 BCAT2

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 High Protein (HPA) HPA →
Cervix Reported tissue cells High Protein (HPA) HPA →
Breast Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Cerebral cortex Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced BCAT2 Western Blot Tips

Deeper troubleshooting and optimisation questions for BCAT2, answered from its protein features.

Which band should guide the blot?
Use 44.3 kDa, the observation attached to the authoritative A06369-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 44.3 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 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 BCAT2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A06369-1 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

BCAT2 Western Blot Reagents

Human/Mouse/Rat-reactive BCAT2 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for BCAT2 using A06369-1; observed band 44.3 kDa
Anti-BCAT2 Antibody Picoband®
Cat # A06369-1

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 O15382
  2. Human Protein Atlas — BCAT2
  3. A06369-1 product record
  4. PMC8945524 — Ruscogenin Alleviates Myocardial Ischemia-Induced Ferroptosis through the Activation of BCAT1/BCAT2 (Antioxidants (Basel, Switzerland), 2022)
  5. PMC5398413 — Genomic deletion of malic enzyme 2 confers collateral lethality in pancreatic cancer (Nature, 2017)
  6. PMC4447349 — Maternal Diabetes Leads to Adaptation in Embryonic Amino Acid Metabolism during Early Pregnancy (PloS one, 2015)