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- Table of Contents
BRAF · Western blot design guide
BRAF Western Blot Planning Guide
Plan a BRAF Western blot around the catalog-observed 84.4 kDa band, image-backed PB9164 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
BRAF 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 | 84.4 kDa observed | |
| Observed band | 84.4 kDa observed | |
| Gel | 8-10% | |
| Positive control | Cerebellum | |
| Negative control | Use knockout/knockdown or orthogonal negative |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 84.4 kDa |
| ⓘ | Localization | Nucleus / Cytoplasm |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated BRAF Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| 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 | PB9164 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 BRAF Western Blot Band Size?
Use the product-observed 84.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 84.4 kDa | Matches the authoritative product WB observation. |
| 84.4 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 BRAF appearancePlan around 84.4 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 84.4 kDa; use this as the primary experimental expectation. |
| Calculated mass | 84.4 kDa from UniProt P15056; 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 PB9164. |
| Situation | Likely cause | Next action |
|---|---|---|
| 84.4 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 BRAF band
PB9164 · BRAFObserved 84.4 kDa · product-specific WB image
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BRAF Western blot
🧪Use Cerebellum as the first positive-control candidate and retain an orthogonal negative control.
Positive control: Cerebellum (High)
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for BRAF
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 |
|---|---|---|---|---|
| Cerebellum | Reported tissue cells | High | Protein (HPA) | HPA → |
| Breast | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Bronchus | Reported tissue cells | High | Protein (HPA) | HPA → |
| Heart muscle | Reported tissue cells | Low | Protein (HPA) | HPA → |
| Skeletal muscle | Reported tissue cells | Low | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
The available HPA profile lists no reliably-negative tissue for BRAF (a broadly-expressed target). Use a BRAF 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 BRAF Western Blot Tips
Deeper troubleshooting and optimisation questions for BRAF, answered from its protein features.
Which band should guide the blot?
Use 84.4 kDa, the observation attached to the authoritative PB9164 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 84.4 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, 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 BRAF Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should PB9164 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
BRAF Western Blot Reagents
Human/Mouse/Rat-reactive BRAF Western blot reagents with authoritative product imagery.
Real WB data
Anti-B Raf/BRAF Antibody Picoband®
Cat # PB9164
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.