CYP2B6 · Western blot design guide

CYP2B6 Western Blot Planning Guide

Plan a CYP2B6 Western blot around the catalog-observed 56.3 kDa band, image-backed A00861-2 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 CYP2B6 (CYP2B6): expected band 56.3 kDa, antibody A00861-2, and guide-derived SDS-PAGE protocol steps
CYP2B6 Western blot protocol sheet — expected band 56.3 kDa, antibody A00861-2, controls and PMC citations. Open the full CYP2B6 WB guide →

CYP2B6 Western Blot Experimental Design Guide

Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running
Expected band 56.3 kDa
Observed band Not reported — verify product WB image
Gel 12-15%
Positive control ⓘ Liver
Negative control ⓘ Adipose tissue
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 56.3 kDa
ⓘ Localization Endoplasmic reticulum membrane / Microsome membrane
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human
Section 1

Real Curated CYP2B6 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateLiver
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
PrimaryA00861-2 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 CYP2B6 Western Blot Band Size?

Use the product-observed 56.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
56.3 kDaMatches the authoritative product WB observation.
56.3 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected CYP2B6 appearancePlan around 56.3 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band56.3 kDa; use this as the primary experimental expectation.
Calculated mass56.3 kDa from UniProt P20813; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00861-2.
Why is my band missing or off?
SituationLikely causeNext action
56.3 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 CYP2B6 Western blot

🧪Use Liver as the first positive-control candidate and Adipose tissue as the HPA Not detected negative candidate.
Positive control: Liver (High)
Negative control: Adipose tissue (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for CYP2B6

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
Liver Reported tissue cells High Protein (HPA) HPA →
Small intestine Reported tissue cells Medium Protein (HPA) HPA →
Kidney Reported tissue cells Low Protein (HPA) HPA →
Duodenum Reported tissue cells Low Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Reported tissue cells Not detected Protein (HPA) HPA →
Adrenal gland Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced CYP2B6 Western Blot Tips

Deeper troubleshooting and optimisation questions for CYP2B6, answered from its protein features.

Which band should guide the blot?
Use 56.3 kDa, the observation attached to the authoritative A00861-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 56.3 kDa expectation.
Which positive control should I start with?
Start with Liver, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Adipose 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 CYP2B6 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00861-2 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

CYP2B6 Western Blot Reagents

Human-reactive CYP2B6 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for CYP2B6 using A00861-2; observed band 56.3 kDa
Anti-Cytochrome P450 2B6/CYP2B6 Antibody Picoband®
Cat # A00861-2

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 P20813
  2. Human Protein Atlas — CYP2B6
  3. A00861-2 product record
  4. PMC2410158 — CYP3A4, CYP2C9 and CYP2B6 expression and ifosfamide turnover in breast cancer tissue microsomes (British journal of cancer, 2004)
  5. PMC13304254 — Pharmacogenetic CYP2B6 variants affect steroid hormone metabolism in human breast cancer cells (British journal of clinical pharmacology, 2026)
  6. PMC10502999 — METTL3 exacerbates insulin resistance in hepatocytes by regulating m(6)A modification of cytochrome P450 2B6 (Nutrition & metabolism, 2023)