SHISA6 / Protein shisa-6 · Western blot design guide

Design a Western Blot for SHISA6

Source-linked SHISA6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SHISA6 WB antibodies. Everything you need to plan the experiment before you commit precious samples.

Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use

Western blot protocol sheet for SHISA6: expected band ~55.8 kDa, hero antibody A14975-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SHISA6** Western blot protocol sheet — expected band ~55.8 kDa, antibody A14975-1, controls and PMC citations. Open the full SHISA6 WB guide →

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SHISA6 Western Blot Experimental Design Guide

Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running

	Expected band	~55.8 kDa
	Observed band	56 kDa
	Gel	5–20% (catalog A14975-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Glycosylated + Phosphorylated
	Caveat	Modification-state controls
	Gene-set association	MSigDB C7 membership
	Isoform	3 isoform(s)

Section 1

Source-Linked SHISA6 Western Blot Protocol Options

The A14975-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	rat brain, mouse brain (catalog A14975-1)
Gel %	5–20% (catalog A14975-1)
Load	50ug; reducing conditions (catalog A14975-1)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A14975-1)
Membrane	Nitrocellulose membrane (catalog A14975-1)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A14975-1)
Primary antibody	A14975-1 · 0.5 μg/mL (catalog A14975-1)
Primary incubation	overnight at 4°C (catalog A14975-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A14975-1)
Secondary incubation	1.5 hour at RT (catalog A14975-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A14975-1)
Detection	ECL (catalog A14975-1)

Section 2

What Is the Expected SHISA6 Western Blot Band Size?

SHISA6 is predicted at 55.8 kDa and observed near 56 kDa; its annotated glycosylation, signal peptide, and isoforms have no demonstrated separate migration pattern.

What am I looking at on my blot?
Band near 56 kDa	Matches the observed SHISA6 band in reducing brain lysates
Band above 56 kDa	Could reflect N-linked glycosylation at Asn32 or Asn59; a shift is unproven
Band below the precursor position	Could reflect signal-peptide cleavage; a separate mature band is unproven
Several bands at different positions	Could include isoforms 1, 2, and 3; distinct migration is unproven

💡Expected SHISA6 appearanceSHISA6 is predicted at 55.8 kDa and observed near 56 kDa in reducing brain lysates; confirm band identity with an independent antibody or SHISA6 loss control.

How each factor affects band size
UniProt predicted mass	55.8 kDa, consistent with the observed band near 56 kDa
N-linked glycosylation at Asn32	May alter apparent size; no visible shift is established
N-linked glycosylation at Asn59	May alter apparent size; no visible shift is established
Signal peptide at residues 1–25	Cleavage may lower mature protein mass relative to the precursor; separate migration is unproven
Splice isoforms 1, 2, and 3	May differ in size; distinct band positions are unestablished

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane SHISA6 may be poorly recovered	Check membrane protein extraction and include a brain lysate positive control
Band higher than expected	N-linked glycosylation or an unrelated antibody target may contribute	Compare glycosidase-treated and untreated samples and verify identity with SHISA6 loss control
Band lower than expected	Signal-peptide cleavage may contribute, but a separate mature band is unproven	Compare with the 56 kDa reference band and verify identity with an independent antibody
Multiple bands	Isoforms 1, 2, and 3 are annotated, but their migration is unknown	Use an independent antibody or SHISA6 loss control to identify SHISA6 bands
Broad smear instead of sharp band	Variable N-linked glycosylation is possible but unverified	Compare glycosidase-treated and untreated samples and check sample quality
Weak or no signal	Low recovery of membrane-associated SHISA6 is possible	Check extraction efficiency and antibody detection with brain lysate

Source: UniProt Q6ZSJ9 Human Protein Atlas product datasheet (A14975-1)

Experiment evidence

Western blot images showing SHISA6 band

A14975-1 · SHISA6

← swipe / scroll for more images →

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

Sample controls for SHISA6 Western blot

🧪For positive controls for SHISA6 in Western blot, you can use an HPA-validated tissue or cell line once one is identified.

Positive control: No high/medium HPA tissue identified

Negative control: Suggested KO / knockdown lysate

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside SHISA6.

⚠️Feasibility: HPA provides no tissue data here, so suitable positive and negative samples cannot be selected from this evidence.

HPA tissue expression evidence for SHISA6

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.

Higher expression tissues · candidate positive controls from IHC

Tissue	Cell type	Level	Evidence	Source
No high/medium HPA tissues identified in the supplied evidence.

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
No lower-expression tissue rows available in the supplied evidence.

Section 3

Advanced SHISA6 Western Blot Tips

Deeper troubleshooting and optimisation questions for SHISA6, answered from its protein features.

How should SHISA6 band migration be interpreted?

Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.

Could SHISA6 isoforms produce different bands?

Isoforms · UniProt lists isoforms 1, 2, and 3. At canonical residue 266, isoforms 2 and 3 replace P with different longer sequences. Consider isoform expression when evaluating multiple bands; the features do not establish where each isoform runs.

Which glycosylation sites matter when interpreting SHISA6 bands?

PTM · UniProt annotates N-linked glycosylation at Asn32 and Asn59. These are canonical UniProt coordinates. Glycosylation may affect migration, but the site annotations alone cannot assign a particular band to a glycosylated form.

UniProt lists phosphoserine at 391, 397, and 409, and phosphothreonine at 433 and 477. These are canonical UniProt coordinates; check the numbering convention before comparing them with antibody documentation. Site annotation alone does not establish a visible band shift.

Does this guide establish induction of SHISA6?

Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.

How should transfer be checked for SHISA6?

Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A14975-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.

How should multiple SHISA6 bands be quantified?

Quantitation · State whether quantitation uses the observed 56 kDa band alone or includes other bands. SHISA6 has three isoforms and annotated glycosylation and phosphorylation sites, so a change in one band cannot by itself establish a change in total SHISA6.

Why does SHISA6 run near 56 kDa?

Interpretation · The observed 56 kDa band is close to the predicted 55.8 kDa. SHISA6 has a signal peptide at residues 1–25 and N-linked glycosylation sites at Asn32 and Asn59, but these annotations alone do not establish a visible shift or explain the small difference.

Check candidate bands against the observed 56 kDa band and the predicted 55.8 kDa mass. Isoforms 2 and 3 differ at canonical residue 266; UniProt also lists N-linked glycosylation at 32 and 59 and five phosphorylation sites. These features provide possibilities, not proof of any unexpected band's identity.

Boster reagents

SHISA6 Western Blot Antibodies

Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.

Real WB data

Western blot analysis of SHISA6 using anti-SHISA6 antibody (A14975-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SHISA6 antigen affinity purified polyclonal antibody (Catalog # A14975-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SHISA6 at approximately 56KD. The expected band size for SHISA6 is at 56KD.

Anti-SHISA6 Antibody Picoband®

Cat # A14975-1

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The catalog reports one anti-SHISA6 antibody, A14975-1, with stated human, mouse, and rat reactivity. Its Western blot image uses rat and mouse brain lysates and shows a band near the expected 56 kDa; human samples are not shown.

Which to pick: A14975-1 is the only listed option. Its WB image provides a starting reference for rat or mouse brain lysates under the reported conditions. Human reactivity is listed, but the supplied image does not demonstrate a human sample.

Source: BosterBio SHISA6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q6ZSJ9.
Human Protein Atlas. SHISA6 tissue expression.
PMC4778035 — target-verified WB comparison
PMC7175376 — target-verified WB comparison