TP53BP1 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 213.6 kDa observed
Observed band 213.6 kDa observed
Gel 4-12% gradient
Positive control ⓘ Adrenal gland
Negative control ⓘ Use knockout/knockdown or orthogonal negative
Important caveats
Reasons your observed band may differ from the expected size.
ⓘCalculated mass213.6 kDa
ⓘLocalizationNucleus / Chromosome
ⓘProcessing / PTMRecord-dependent
ⓘReactivityHuman / Mouse / Rat
Section 1

Real Curated TP53BP1 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateAdrenal gland
Gel %4-12% gradient
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA00397-1 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 TP53BP1 Western Blot Band Size?

Use the product-observed 213.6 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
213.6 kDaMatches the authoritative product WB observation.
213.6 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected TP53BP1 appearancePlan around 213.6 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band213.6 kDa; use this as the primary experimental expectation.
Calculated mass213.6 kDa from UniProt Q12888; retain as context.
Gel selection4-12% gradient; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A00397-1.
Why is my band missing or off?
SituationLikely causeNext action
213.6 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 TP53BP1 Western blot

🧪Use Adrenal gland as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Adrenal gland (High)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for TP53BP1

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 →
Caudate Reported tissue cells High Protein (HPA) HPA →
Appendix Reported tissue cells High Protein (HPA) HPA →
Bone marrow Reported tissue cells Medium Protein (HPA) HPA →
Liver Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

The available HPA profile lists no reliably-negative tissue for TP53BP1 (a broadly-expressed target). Use a TP53BP1 knockdown (siRNA/shRNA) or knockout lysate as the negative control and confirm signal loss versus the parental lysate.
Tissue Cell type Level Evidence Source
Section 3

Advanced TP53BP1 Western Blot Tips

Deeper troubleshooting and optimisation questions for TP53BP1, answered from its protein features.

Which band should guide the blot?
Use 213.6 kDa, the observation attached to the authoritative A00397-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 213.6 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 4-12% gradient consistently across the quick facts, protocol table, and poster.
What transfer method to use for TP53BP1 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A00397-1 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

TP53BP1 Western Blot Reagents

Human/Mouse/Rat-reactive TP53BP1 Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for TP53BP1 using A00397-1; observed band 213.6 kDa
Anti-TP53BP1 Antibody Picoband®
Cat # A00397-1
Real WB dataWestern blot validation image for TP53BP1 using A00397; observed band 213.6 kDa
Anti-53BP1/TP53BP1 Antibody Picoband®
Cat # A00397
Real WB dataWestern blot validation image for TP53BP1 using PA1976; observed band 213.6 kDa
Anti-53BP1/TP53BP1 Antibody Picoband®
Cat # PA1976

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.