This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
RPTOR · Western blot design guide
RPTOR Western Blot Planning Guide
Plan a RPTOR Western blot around the catalog-observed 149 kDa band, image-backed A01463-3 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
RPTOR 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 | 149 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 8-10% | |
| Positive control | Adrenal gland | |
| Negative control | Fallopian tube |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 149 kDa |
| ⓘ | Localization | Lysosome membrane / Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated RPTOR 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 | Wet/tank, extended transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A01463-3 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 RPTOR Western Blot Band Size?
Use the product-observed 149 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 149 kDa | Matches the authoritative product WB observation. |
| 149 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 RPTOR appearancePlan around 149 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 149 kDa; use this as the primary experimental expectation. |
| Calculated mass | 149 kDa from UniProt Q8N122; 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 A01463-3. |
| Situation | Likely cause | Next action |
|---|---|---|
| 149 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 RPTOR band
A01463-3 · RPTOR
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RPTOR Western blot
🧪Use Adrenal gland as the first positive-control candidate and Fallopian tube as the HPA Not detected negative candidate.
Positive control: Adrenal gland (High)
Negative control: Fallopian tube (Not detected)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for RPTOR
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 |
|---|---|---|---|---|
| Fallopian tube | Reported tissue cells | Not detected | Protein (HPA) | HPA → |
Section 3
Advanced RPTOR Western Blot Tips
Deeper troubleshooting and optimisation questions for RPTOR, answered from its protein features.
Which band should guide the blot?
Use 149 kDa, the observation attached to the authoritative A01463-3 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 149 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 Fallopian tube as an orthogonal HPA Not detected candidate.
Which gel should I use?
Use 8-10% consistently across the quick facts, protocol table, and poster.
What transfer method to use for RPTOR Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A01463-3 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
Compare only the supplied target-verified records: PMC13054871, PMC9344631.
Boster reagents
RPTOR Western Blot Reagents
Human/Mouse/Rat-reactive RPTOR Western blot reagents with authoritative product imagery.
Real WB data
Anti-Raptor/RPTOR Antibody Picoband®
Cat # A01463-3
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 Q8N122
- Human Protein Atlas — RPTOR
- A01463-3 product record
- PMC13054871 — Site-Specific and Quantitative O-GlcNAc Proteomics for Hepatocellular Carcinoma.
- PMC9344631 — AKT phosphorylation as a predictive biomarker for PI3K/mTOR dual inhibition-induced proteolytic cleavage of mTOR companion proteins in small cell lung cancer.