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- Table of Contents
CASK · Western blot design guide
CASK Western Blot Planning Guide
Plan a CASK Western blot around the catalog-observed 20.3 kDa band, image-backed A02468-2 evidence, HPA controls, and verified protocol records.
Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
CASK Western Blot Experimental Design Guide
Expected bands, documented protocol parameters, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | 20.3 kDa | |
| Observed band | Not reported — verify product WB image | |
| Gel | 12-15% | |
| Positive control | Small intestine | |
| Negative control | Target knockdown/knockout |
Important caveats
Reasons your observed band may differ from the expected size.
| ⓘ | Calculated mass | 20.3 kDa |
| ⓘ | Localization | Secreted |
| ⓘ | Processing / PTM | Record-dependent |
| ⓘ | Reactivity | Human / Mouse / Rat |
Section 1
Real Curated CASK Western Blot Protocols
Start with the molecular-weight rule, then compare verified publication-derived conditions.
| Sample / lysate | Appendix |
| Gel % | 12-15% |
| Load | 20-30 µg total protein per lane |
| Transfer | Semi-dry, short transfer |
| Membrane | 0.45 µm PVDF |
| Blocking | 5% non-fat milk or 5% BSA in TBST |
| Primary | A02468-2 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 CASK Western Blot Band Size?
Use the product-observed 20.3 kDa band as the primary planning value and retain the UniProt calculated mass as context.
| 20.3 kDa | Matches the authoritative product WB observation. |
| 20.3 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 CASK appearancePlan around 20.3 kDa and keep the calculated mass as supporting context.
| Catalog-observed band | 20.3 kDa; use this as the primary experimental expectation. |
| Calculated mass | 20.3 kDa from UniProt P07498; retain as context. |
| Gel selection | 12-15%; shared with the recommended protocol and poster. |
| Specificity check | Compare the lead HPA positive and negative controls with A02468-2. |
| Situation | Likely cause | Next action |
|---|---|---|
| 20.3 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 CASK band
A02468-2 · CASK
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CASK Western blot
🧪Use Small intestine as the first positive-control candidate; no defensible HPA Not detected tissue was available, so use a target knockdown/knockout negative control.
Positive control: Small intestine (High)
Negative control: Target knockdown/knockout
HPA protein score determines control status; other expression data is supporting context only.
HPA tissue expression evidence for CASK
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 |
|---|---|---|---|---|
| Small intestine | Reported tissue cells | High | Protein (HPA) | HPA → |
| Colon | Reported tissue cells | High | Protein (HPA) | HPA → |
| Appendix | Reported tissue cells | High | Protein (HPA) | HPA → |
| Duodenum | Reported tissue cells | High | Protein (HPA) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|
Section 3
Advanced CASK Western Blot Tips
Deeper troubleshooting and optimisation questions for CASK, answered from its protein features.
Which band should guide the blot?
Use 20.3 kDa, the observation attached to the authoritative A02468-2 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 20.3 kDa expectation.
Which positive control should I start with?
Start with Small intestine, 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 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for CASK Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A02468-2 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
CASK Western Blot Reagents
Human/Mouse/Rat-reactive CASK Western blot reagents with authoritative product imagery.
Real WB data
Anti-CASK Antibody Picoband®
Cat # A02468-2
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 P07498
- Human Protein Atlas — CASK
- A02468-2 product record
- PMC7235163 — CASK, the Soluble Glomerular Permeability Factor, Is Secreted by Macrophages in Patients With Recurrent Focal and Segmental Glomerulo-Sclerosis (Frontiers in immunology, 2020)
- PMC4220808 — Mutated CaV2.1 channels dysregulate CASK/P2X3 signaling in mouse trigeminal sensory neurons of R192Q Cacna1a knock-in mice (Molecular pain, 2013)
- PMC4818453 — X-linked intellectual disability gene CASK regulates postnatal brain growth in a non-cell autonomous manner (Acta neuropathologica communications, 2016)