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- Table of Contents
SETDB2 · Western blot design guide
SETDB2 Western Blot Planning Guide
Plan a SETDB2 Western blot around the catalog-observed 81.9 kDa band, image-backed A08290-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
SETDB2 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 | 81.9 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Adrenal gland | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 81.9 kDa |
| ⓘ | Localization | Nucleus / Chromosome |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated SETDB2 Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Adrenal gland |
| Gel % | 8-10% |
| 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 | A08290-2 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 2
What Is the Expected SETDB2 Western Blot Band Size?
Use the product-observed 81.9 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 81.9 kDa | Matches the authoritative product WB observation. |
| 81.9 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 SETDB2 appearancePlan around 81.9 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 81.9 kDa; use this as the primary experimental expectation. |
| Calculated mass | 81.9 kDa from UniProt Q96T68; retain as context. |
| Gel selection | 8-10%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A08290-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 81.9 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. |
Experiment evidence
Western blot images showing SETDB2 band
A08290-2 · SETDB2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SETDB2 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 SETDB2
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
| 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 |
|---|
Section 3
Advanced SETDB2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SETDB2, answered from its protein features.
Which band should guide the blot?
Use 81.9 kDa, the observation attached to the authoritative A08290-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 81.9 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 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for SETDB2 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A08290-2 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
SETDB2 Western Blot Reagents
Human/Mouse/Rat-reactive SETDB2 Western blot reagents with authoritative product imagery.
Real WB data
Anti-CLLD8/SETDB2 Picoband® Antibody
Cat # A08290-2
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
- UniProt Q96T68
- Human Protein Atlas — SETDB2
- A08290-2 product record
- PMC7294945 — SETDB2 promoted breast cancer stem cell maintenance by interaction with and stabilization of ΔNp63α protein (International journal of biological sciences, 2020)
- PMC5341872 — Oncogenic roles of the SETDB2 histone methyltransferase in gastric cancer (Oncotarget, 2016)
- PMC10909981 — SETDB2 interacts with BUBR1 to induce accurate chromosome segregation independently of its histone methyltransferase activity (FEBS open bio, 2024)