SLC12A5 · Western blot design guide

SLC12A5 Western Blot Planning Guide

Plan a SLC12A5 Western blot around the catalog-observed 126.2 kDa band, image-backed A04629-1 evidence, HPA controls, and verified protocol records.

Evidence assembled July 2026 · For research use; verify linked source records and product datasheet before use
Western blot protocol sheet for SLC12A5 (SLC12A5): expected band 126.2 kDa, antibody A04629-1, and guide-derived SDS-PAGE protocol steps
SLC12A5 Western blot protocol sheet — expected band 126.2 kDa, antibody A04629-1, controls and PMC citations. Open the full SLC12A5 WB guide →

SLC12A5 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 126.2 kDa
Observed band Not reported — verify product WB image
Gel 8-10%
Positive control ⓘ Cerebellum
Negative control ⓘ Appendix
Important caveats
Reasons your observed band may differ from the expected size.
ⓘ Calculated mass 126.2 kDa
ⓘ Localization Cell membrane / Cell projection, dendrite
ⓘ Processing / PTM Record-dependent
ⓘ Reactivity Human / Mouse / Rat
Section 1

Real Curated SLC12A5 Western Blot Protocols

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

Recommended Western blot protocol parameters
Sample / lysateCerebellum
Gel %8-10%
Load20-30 µg total protein per lane
TransferWet/tank, extended transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA04629-1 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 SLC12A5 Western Blot Band Size?

Use the product-observed 126.2 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
126.2 kDaMatches the authoritative product WB observation.
126.2 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected SLC12A5 appearancePlan around 126.2 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band126.2 kDa; use this as the primary experimental expectation.
Calculated mass126.2 kDa from UniProt Q9H2X9; retain as context.
Gel selection8-10%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A04629-1.
Why is my band missing or off?
SituationLikely causeNext action
126.2 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 SLC12A5 Western blot

🧪Use Cerebellum as the first positive-control candidate and Appendix as the HPA Not detected negative candidate.
Positive control: Cerebellum (Medium)
Negative control: Appendix (Not detected)
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for SLC12A5

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

TissueCell typeLevelEvidenceSource
Cerebellum Reported tissue cells Medium Protein (HPA) HPA →
Cerebral cortex Reported tissue cells Medium Protein (HPA) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Appendix Reported tissue cells Not detected Protein (HPA) HPA →
Bone marrow Reported tissue cells Not detected Protein (HPA) HPA →
Section 3

Advanced SLC12A5 Western Blot Tips

Deeper troubleshooting and optimisation questions for SLC12A5, answered from its protein features.

Which band should guide the blot?
Use 126.2 kDa, the observation attached to the authoritative A04629-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 126.2 kDa expectation.
Which positive control should I start with?
Start with Cerebellum, the lead HPA protein-expression candidate.
Which negative control is defensible?
Use Appendix 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 SLC12A5 Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A04629-1 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

SLC12A5 Western Blot Reagents

Human/Mouse/Rat-reactive SLC12A5 Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for SLC12A5 using A04629-1; observed band 126.2 kDa
Anti-KCC2/SLC12A5 Antibody Picoband®
Cat # A04629-1

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

  1. UniProt Q9H2X9
  2. Human Protein Atlas — SLC12A5
  3. A04629-1 product record
  4. PMC10134347 — SLC12A5 promotes hepatocellular carcinoma growth and ferroptosis resistance by inducing ER stress and cystine transport changes (Cancer medicine, 2023)
  5. PMC4819609 — Increased expression of Solute carrier family 12 member 5 via gene amplification contributes to tumour progression and metastasis and associates with poor survival in colorectal cancer (Gut, 2016)