CYP7A1 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 57.7 kDa observed
Observed band 57.7 kDa observed
Gel 12-15%
Positive control ⓘ No defensible HPA tissue; use knockout/knockdown
Negative control ⓘ Use knockout/knockdown or orthogonal negative
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass57.7 kDa
ⓘLocalizationEndoplasmic reticulum membrane / Microsome membrane
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Rat
Section 1

Real Curated CYP7A1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateKnown-positive human lysate
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
PrimaryRP1079 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 CYP7A1 Western Blot Band Size?

Use the product-observed 57.7 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
57.7 kDaMatches the authoritative product WB observation.
57.7 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected CYP7A1 appearancePlan around 57.7 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band57.7 kDa; use this as the primary experimental expectation.
Calculated mass57.7 kDa from UniProt P22680; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with RP1079.
Why is my band missing or off?
SituationLikely causeNext action
57.7 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 CYP7A1 Western blot

🧪No defensible HPA positive control was available; do not invent one.
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for CYP7A1

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

Undetected expression · recommended negative controls

The Human Protein Atlas exposes no structured IHC tissue profile for CYP7A1, so tissue-level positive/negative controls cannot be listed here. Use a documented CYP7A1-positive lysate (recombinant CYP7A1 or a known-expressing line) as the positive control and a CYP7A1 knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
Tissue Cell type Level Evidence Source
Section 3

Advanced CYP7A1 Western Blot Tips

Deeper troubleshooting and optimisation questions for CYP7A1, answered from its protein features.

Which band should guide the blot?
Use 57.7 kDa, the observation attached to the authoritative RP1079 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 57.7 kDa expectation.
Which positive control should I start with?
No defensible positive tissue was supplied; do not invent one.
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 CYP7A1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should RP1079 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

CYP7A1 Western Blot Reagents

Human/Rat-reactive CYP7A1 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for CYP7A1 using RP1079; observed band 57.7 kDa
Anti-CYP7A1 Antibody Picoband®
Cat # RP1079
Real WB dataWestern blot validation image for CYP7A1 using A01601; observed band 57.7 kDa
Anti-CYP7A1 Antibody Picoband®
Cat # A01601

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.