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- Table of Contents
SHANK2 / SH3 and multiple ankyrin repeat domains protein 2 · Western blot design guide
Design a Western Blot for SHANK2
Real validated SHANK2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SHANK2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
SHANK2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~201.3 kDa | |
| Observed band | ~200 kDa | |
| Gel | 8% (catalog A03782-2) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
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 | 4 isoform(s) |
Section 1
Real Curated SHANK2 Western Blot Protocols
The A03782-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, mouse brain (catalog A03782-2) |
| Gel % | 8% (catalog A03782-2) |
| Load | 30 ug; reducing conditions (catalog A03782-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03782-2) |
| Membrane | nitrocellulose membrane (catalog A03782-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03782-2) |
| Primary antibody | A03782-2 · 0.5 μg/mL (catalog A03782-2) |
| Primary incubation | overnight at 4°C (catalog A03782-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03782-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03782-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03782-2) |
| Detection | ECL (catalog A03782-2) |
Section 2
What Is the Expected SHANK2 Western Blot Band Size?
SHANK2 is predicted at 201.3 kDa and observed at ~200 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~200 kDa | Empirical SHANK2 band, close to the 201.3 kDa predicted mass |
| Band near 201.3 kDa | Consistent with the predicted SHANK2 size; confirm identity with controls |
| Several bands at different masses | Could include SHANK2 splice isoforms 1, 2, 3, and 4; their migration is unspecified |
| Faint band in a soluble fraction | SHANK2 also localizes to membranes and the postsynaptic density |
💡Expected SHANK2 appearanceUniProt predicts SHANK2 at 201.3 kDa, and antibody QC reports a band at ~200 kDa; confirm its identity with a positive control and an independent specificity control.
| UniProt predicted mass | 201.3 kDa predicted; the empirical band is ~200 kDa |
| O-linked GlcNAc at Thr1667 | Adds one monosaccharide; a visible shift is not established |
| Splice isoform 1 | May differ in size from other isoforms; mass and migration are unspecified |
| Splice isoform 2 | May differ in size from other isoforms; mass and migration are unspecified |
| Splice isoform 3 | May differ in size from other isoforms; mass and migration are unspecified |
| Splice isoform 4 | May differ in size from other isoforms; mass and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SHANK2 may be poorly represented in the sampled lysate despite its synaptic localization | Check a brain lysate positive control and consider a synapse-enriched fraction |
| Band higher than expected | A different isoform or nonspecific binding is possible; the cause is unestablished | Compare with the ~200 kDa control band and verify identity independently |
| Band lower than expected | An isoform or unrelated band is possible; no fragment size is established | Compare with the ~200 kDa control band and check with a second antibody or depletion control |
| Multiple bands | SHANK2 has four splice isoforms, but their band positions are unknown | Use an independent specificity control to identify SHANK2 bands |
| Weak or no signal | SHANK2 abundance in the sampled fraction or transfer of the large protein may limit detection | Check a brain lysate positive control and transfer near 200 kDa |
Experiment evidence
Western blot images showing SHANK2 band
A03782-2 · SHANK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SHANK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SHANK2 in Western blot, you can use appendix tissue lysate, which has medium HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA reports only medium expression in the positive tissue, so SHANK2 detection may require sufficient lysate.
HPA tissue expression evidence for SHANK2
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 |
|---|---|---|---|---|
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SHANK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SHANK2, answered from its protein features.
How should SHANK2 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 SHANK2 isoforms produce different bands?
Isoforms · Yes. UniProt lists four isoforms. Isoform 2 lacks reference residues 870–1849 and should be substantially shorter. Isoforms 1, 2, and 4 have altered N-terminal sequences; isoform 4 also lacks residues 763–769. Check whether the antibody epitope is retained before assigning a band to an isoform.
Which phosphorylation sites matter when interpreting SHANK2 bands?
PTM · UniProt lists phosphoserines at reference positions 831, 960, 1099, 1709, and 1713, and phosphothreonines at 860 and 1278. These are UniProt reference coordinates; antibody or paper numbering may differ. Phosphorylation could affect detection or migration, but these entries alone do not establish a visible band shift. Reference residue 860 is replaced in isoform 2.
UniProt lists O-linked GlcNAc on threonine 1667 in reference-sequence numbering. Isoform 2 lacks the reference region containing this site. The annotation alone cannot establish whether glycosylation changes the apparent band position.
UniProt lists O-linked GlcNAc on threonine 1667 in reference-sequence numbering. Isoform 2 lacks the reference region containing this site. The annotation alone cannot establish whether glycosylation changes the apparent band position.
Does this guide establish induction of SHANK2?
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 SHANK2 Western blot?
Transfer · SHANK2 is predicted at 201.3 kDa. Choose a transfer setup suited to proteins around 200 kDa, and confirm transfer in that region with a total-protein stain or another transfer check.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03782-2 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 SHANK2 bands be quantified?
Quantitation · Define whether the measurement targets full-length SHANK2 or multiple isoforms before selecting bands. Because isoform 2 lacks reference residues 870–1849, an antibody against that region would exclude it. Quantify only bands consistent with the antibody's recognized sequence and report which bands were included.
How should the observed SHANK2 band compare with predicted mass?
Interpretation · The predicted mass is 201.3 kDa, close to the reported apparent band near 200 kDa. Use that region as a starting point for identifying full-length SHANK2; the listed modifications do not establish a visible shift.
First compare its position and antibody epitope with the four annotated isoforms. Isoform 2 lacks reference residues 870–1849; isoform 4 lacks 763–769. The listed phosphorylation and O-GlcNAc sites are possible variables, but their presence does not identify an unexpected band or prove a mobility shift.
First compare its position and antibody epitope with the four annotated isoforms. Isoform 2 lacks reference residues 870–1849; isoform 4 lacks 763–769. The listed phosphorylation and O-GlcNAc sites are possible variables, but their presence does not identify an unexpected band or prove a mobility shift.
Boster reagents
SHANK2 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
Anti-SHANK2 Antibody Picoband®
Cat # A03782-2
The catalog reports one anti-SHANK2 antibody for Western blot, A03782-2, with stated human, mouse, and rat reactivity. Its WB image uses rat and mouse brain lysates; the reported band is approximately 200 kDa versus an expected 159 kDa.
Which to pick: A03782-2 is the only listed option. Its WB image supports use in rat and mouse brain lysates under the reported conditions. Human reactivity is listed, but no human WB sample is shown in the supplied caption.
Source: BosterBio SHANK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UPX8.
- Human Protein Atlas. SHANK2 tissue expression.
- PMC10265811 — target-verified WB comparison
- PMC5302174 — target-verified WB comparison
- PMC6018399 — target-verified WB comparison