SEMA6A · Western blot design guide

SEMA6A Western Blot Planning Guide

Plan a SEMA6A Western blot around the catalog-observed 114.4 kDa band, image-backed A06967-1 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 SEMA6A (SEMA6A): expected band 114.4 kDa, antibody A06967-1, and PMC-cited SDS-PAGE protocol steps — SEMA6A Western blot protocol sheet — expected band 114.4 kDa, antibody A06967-1, controls and PMC citations. Open the full SEMA6A WB guide →

Navigate to: Protocols Band Controls Antibodies

SEMA6A 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	114.4 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	114.4 kDa
ⓘ	Localization	Cell membrane
ⓘ	Processing / PTM	Record-dependent
ⓘ	Reactivity	Human / Mouse / Rat

Section 1

Real Curated SEMA6A Western Blot Protocols

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

Recommended Western blot protocol parameters
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	A06967-1 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 SEMA6A Western Blot Band Size?

Use the product-observed 114.4 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
114.4 kDa	Matches the authoritative product WB observation.
114.4 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 SEMA6A appearancePlan around 114.4 kDa and keep the calculated mass as supporting context.

How each factor affects band size
Catalog-observed band	114.4 kDa; use this as the primary experimental expectation.
Calculated mass	114.4 kDa from UniProt Q9H2E6; 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 A06967-1.

Why is my band missing or off?
Situation	Likely cause	Next action
114.4 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.

Sources: UniProt Q9H2E6 Human Protein Atlas product datasheet (A06967-1)

Experiment evidence

Western blot images showing SEMA6A band

A06967-1 · SEMA6A

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for SEMA6A 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 SEMA6A

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 SEMA6A Western Blot Tips

Deeper troubleshooting and optimisation questions for SEMA6A, answered from its protein features.

Which band should guide the blot?

Use 114.4 kDa, the observation attached to the authoritative A06967-1 WB record.

How should calculated mass be interpreted?

Treat the UniProt calculated mass as context; it does not replace the catalog-observed 114.4 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 SEMA6A Western blot?

Use the transfer method in the recommended protocol and verify transfer before blocking.

How should A06967-1 be started?

Start at the linked datasheet condition and run a three-point primary-antibody dilution test.

Which publication-derived protocols can I compare?

Compare only the supplied target-verified records: PMC9899268, PMC6281666, PMC6746772.

Boster reagents

SEMA6A Western Blot Reagents

Human/Mouse/Rat-reactive SEMA6A Western blot reagents with authoritative product imagery.

Real WB data Western blot validation image for SEMA6A using A06967-1; observed band 114.4 kDa

Anti-Sema6A Antibody Picoband®

Cat # A06967-1

View product →

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.