SHISA7 / Protein shisa-7 · Western blot design guide

Design a Western Blot for SHISA7

Source-linked SHISA7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SHISA7 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 SHISA7: expected band ~56.2 kDa, hero antibody A18878-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SHISA7** Western blot protocol sheet — expected band ~56.2 kDa, antibody A18878-1, controls and PMC citations. Open the full SHISA7 WB guide →

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SHISA7 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	~56.2 kDa
	Observed band	70 kDa
	Gel	5–20% (catalog A18878-1)
	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	1 isoform(s)

Section 1

Source-Linked SHISA7 Western Blot Protocol Options

The A18878-1 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 A18878-1)
Gel %	5–20% (catalog A18878-1)
Load	50ug; reducing conditions (catalog A18878-1)
Transfer	a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A18878-1)
Membrane	Nitrocellulose membrane (catalog A18878-1)
Blocking	5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A18878-1)
Primary antibody	A18878-1 · 0.5 μg/mL (catalog A18878-1)
Primary incubation	overnight at 4°C (catalog A18878-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A18878-1)
Secondary incubation	1.5 hour at RT (catalog A18878-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A18878-1)
Detection	ECL (catalog A18878-1)

Section 2

What Is the Expected SHISA7 Western Blot Band Size?

SHISA7 is predicted at 56.2 kDa and observed near 70 kDa in reducing brain lysates; the cause of the difference is not established.

What am I looking at on my blot?
Band near 70 kDa	Empirical SHISA7 band in reducing brain lysates; confirm identity with antibody controls
Band near 56.2 kDa	Near the predicted precursor mass; identity requires confirmation
Band below the precursor	Signal-peptide cleavage could yield a smaller mature protein
Band above the predicted mass	N-linked glycosylation at Asn23 or Asn59 may affect migration; the cause of the 70 kDa band is unproven

💡Expected SHISA7 appearanceSHISA7 has a predicted precursor mass of 56.2 kDa, while reducing brain lysates show an empirical band near 70 kDa; its migration difference is unexplained, so confirm band identity with ordinary antibody controls.

How each factor affects band size
Predicted precursor mass	56.2 kDa is the sequence-based reference, not the observed migration
N-linked glycosylation at Asn23	May increase apparent mass if occupied; the size effect is unknown
N-linked glycosylation at Asn59	May increase apparent mass if occupied; the size effect is unknown
Signal peptide at residues 1–19	Cleavage can make the mature protein smaller than the precursor; its migration is unknown

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Membrane-associated SHISA7 may be poorly recovered in a soluble lysate	Check membrane extraction and include a positive brain lysate control
Band higher than expected	N-linked glycosylation may affect migration, but the 70 kDa difference is unexplained	Compare with the 70 kDa brain-lysate reference and check identity using antibody controls
Band lower than expected	Signal-peptide cleavage could reduce size, but the resulting band size is unknown	Check epitope recognition and compare with a positive control
Multiple bands	Different glycosylation or processing states are possible but unverified	Use glycosidase treatment and antibody specificity controls to identify the bands
Weak or no signal	Postsynaptic membrane localization may limit recovery	Enrich the membrane fraction and verify transfer with a positive control

Source: UniProt A6NL88 Human Protein Atlas product datasheet (A18878-1)

Experiment evidence

Western blot images showing SHISA7 band

A18878-1 · SHISA7

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

Sample controls for SHISA7 Western blot

🧪For positive controls for SHISA7 in Western blot, you can use no HPA-IHC candidate positive sample because HPA provides no expression data.

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 the samples.

⚠️Feasibility: HPA provides no tissue controls, and SHISA7 is a postsynaptic membrane protein, so a clear signal may require synapse-enriched material.

HPA tissue expression evidence for SHISA7

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 SHISA7 Western Blot Tips

Deeper troubleshooting and optimisation questions for SHISA7, answered from its protein features.

How should SHISA7 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 SHISA7 isoforms explain multiple bands?

Isoforms · The supplied features list one isoform and no alternative sequence. They therefore provide no basis for assigning additional bands to SHISA7 isoforms. Verify unexpected bands before interpreting them as SHISA7.

Could glycosylation affect the SHISA7 band?

PTM · UniProt lists N-linked glycosylation at Asn23 and Asn59, using canonical sequence coordinates. Compare treated and untreated samples if testing glycosylation; a change would need experimental confirmation. The listed sites alone do not establish a visible shift.

UniProt lists phosphoserine at position 417 and phosphothreonine at position 512. These are canonical sequence coordinates and may differ from paper or antibody numbering. Their presence does not establish a visible band shift; compare matched samples experimentally if phosphorylation is the question.

Does this guide establish induction of SHISA7?

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.

What transfer method to use for SHISA7 Western blot?

Transfer · SHISA7 is a single-pass membrane protein with an observed band near 70 kDa. Check that your transfer conditions retain and transfer protein at that apparent size, using a membrane stain or total-protein check. The supplied features do not specify a transfer method.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A18878-1 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 SHISA7 bands be quantified?

Quantitation · Measure the same verified SHISA7 band across samples and normalize for loading. Keep sample preparation consistent: SHISA7 is a postsynaptic membrane protein localized at GABAergic inhibitory synapses, so differences in membrane or synaptic material can affect the measured signal.

Why might SHISA7 appear at 70 kDa instead of 56.2 kDa?

Interpretation · The supplied Western blots show an apparent 70 kDa band, while the predicted mass is 56.2 kDa. SHISA7 has a signal peptide at residues 1–19 and two N-linked glycosylation sites, but these features alone do not establish the cause of the difference. Use the observed band as a reference and verify its identity experimentally.

The features list one isoform, a signal peptide at residues 1–19, N-linked sites at Asn23 and Asn59, and phosphorylation at Ser417 and Thr512. These suggest properties to investigate, but none identifies an unexpected band by itself. Compare its behavior with the established 70 kDa band and verify its identity.

Boster reagents

SHISA7 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 SHISA7 using anti-SHISA7 antibody (A18878-1). 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-SHISA7 antigen affinity purified polyclonal antibody (Catalog # A18878-1) 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 SHISA7 at approximately 70KD. The expected band size for SHISA7 is at 70KD.

Anti-SHISA7 Antibody Picoband®

Cat # A18878-1

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The catalog reports one anti-SHISA7 antibody for Western blot, A18878-1, with stated human, mouse, and rat reactivity. Its WB image shows an approximately 70 kDa band in rat and mouse brain lysates; the supplied evidence does not show a human sample.

Which to pick: A18878-1 is the only listed option. Its WB image documents rat and mouse brain lysates at 50 µg per lane using 0.5 µg/mL primary antibody. Human reactivity is listed, but no human WB sample is shown.

Source: BosterBio SHISA7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry A6NL88.
Human Protein Atlas. SHISA7 tissue expression.
PMC5737659 — target-verified WB comparison