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SHANK1 / SH3 and multiple ankyrin repeat domains protein 1 · Western blot design guide
Design a Western Blot for SHANK1
Source-linked SHANK1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SHANK1 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
SHANK1 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 | ~225 kDa | |
| Observed band | ~220–250 kDa | |
| Gel | 8% (catalog A05690-2) | |
| Positive control | Cerebellum (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 | Phosphorylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SHANK1 Western Blot Protocol Options
The A05690-2 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 A05690-2) |
| Gel % | 8% (catalog A05690-2) |
| Load | 30 ug; reducing conditions (catalog A05690-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05690-2) |
| Membrane | nitrocellulose membrane (catalog A05690-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05690-2) |
| Primary antibody | A05690-2 · 0.5 μg/mL (catalog A05690-2) |
| Primary incubation | overnight at 4°C (catalog A05690-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05690-2) |
| Secondary incubation | 1.5 hour at RT (catalog A05690-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05690-2) |
| Detection | ECL (catalog A05690-2) |
Section 2
What Is the Expected SHANK1 Western Blot Band Size?
SHANK1 is predicted at 225 kDa and observed at ~220–250 kDa in reducing brain lysates; the cause of the range is not established.
| Band at ~220–250 kDa | Empirical SHANK1 band in brain lysate; confirm identity with antibody controls |
| Band near 225 kDa | Consistent with the predicted SHANK1 mass |
| Several bands of different sizes | Could reflect isoforms 1, 2, and 3; their migration is not established |
| Higher band under conditions that preserve complexes | Could reflect SHANK1 homomultimerization via its SAM domain |
| Weak band in whole-cell lysate | SHANK1 localization to the postsynaptic density may limit its abundance in the sample |
💡Expected SHANK1 appearanceSHANK1 has a predicted mass of 225 kDa and an empirical band at ~220–250 kDa in reducing brain lysates; confirm band identity with antibody and sample controls.
| Predicted SHANK1 mass | 225 kDa predicted; the empirical brain-lysate band is ~220–250 kDa |
| Splice isoform 1 | May differ in size from other isoforms; its individual mass is not supplied |
| Splice isoform 2 | May differ in size from other isoforms; its individual mass is not supplied |
| Splice isoform 3 | May differ in size from other isoforms; its individual mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SHANK1 is localized to the postsynaptic density and may be scarce in the sampled lysate | Check a brain lysate positive control and consider synaptic enrichment |
| Band higher than expected | SHANK1 may homomultimerize via its SAM domain, although persistence during electrophoresis is unproven | Compare reducing and nonreducing samples and verify band identity |
| Band lower than expected | An alternative splice isoform could differ in size, but isoform masses are unavailable | Check antibody specificity and compare samples with a validated positive control |
| Multiple bands | Isoforms 1, 2, and 3 are annotated, but distinct blot bands are not established | Verify which bands are SHANK1 with antibody specificity controls |
| Weak or no signal | SHANK1 may be poorly represented in a sample lacking postsynaptic material | Run a brain lysate positive control and check sample loading |
Experiment evidence
Western blot images showing SHANK1 band
A05690-2 · SHANK1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SHANK1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SHANK1 in Western blot, you can use cerebellum lysate, a high-expression HPA tissue.
Positive control: Cerebellum (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: SHANK1 is associated with the postsynaptic density, so signal in whole-tissue lysate may be diluted.
HPA tissue expression evidence for SHANK1
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Skin | fibroblasts | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SHANK1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SHANK1, answered from its protein features.
What should be checked when extra SHANK1 bands appear?
Band shift · Compare bands with the 225 kDa predicted mass and the annotated isoforms. Isoform 2 has a large N-terminal deletion, while isoform 3 lacks nine residues in canonical UniProt coordinates. Check antibody epitope coverage and whether smaller bands could reflect degradation. The feature list alone cannot identify an unexpected band.
How might SHANK1 isoforms affect band interpretation?
Isoforms · Three isoforms are listed. In canonical UniProt coordinates, isoform 2 lacks residues 1–613 and replaces residues 614–654; isoform 3 lacks residues 646–654. An antibody against residues 1–613 would miss isoform 2. Check the antibody epitope before assigning a smaller band to an isoform.
Which SHANK1 phosphorylation sites matter for phospho-specific detection?
PTM · Annotated sites are Tyr186 and Ser540, Ser671, Ser791, Ser890, Ser1287, and Ser1436 in the supplied UniProt sequence. Match the antibody’s site and numbering convention to these coordinates. These annotations alone do not predict a visible band shift.
Does this guide establish induction of SHANK1?
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 SHANK1 Western blot?
Transfer · SHANK1 is predicted at 225 kDa, so verify that the transfer captures proteins in this high-mass range. A wet transfer is a practical starting point; optimize transfer conditions and check the membrane and post-transfer gel for incomplete transfer.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05690-2 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 can SHANK1 bands be quantified consistently?
Quantitation · Choose an antibody epitope shared by the isoforms you intend to measure, or quantify isoform bands separately. Keep the same band boundaries across samples and confirm the signal is within a usable measurement range. The annotated isoform differences make antibody coverage important for interpreting total SHANK1 signal.
How should a 220–250 kDa SHANK1 band compare with predicted mass?
Interpretation · Canonical SHANK1 is predicted at 225 kDa, within the observed 220–250 kDa range. Its annotated modifications and isoforms may affect migration, but their presence alone does not establish the cause of any apparent mass difference.
The supplied UniProt coordinates include omega-N-methylarginine at 544, 950, 1051, 1090, 1101, 1423, 1895, 2016, 2036, and 2074, plus asymmetric dimethylarginine at 1253. Confirm which isoform an antibody detects before interpreting a band; annotated methylation alone does not explain its apparent mass.
The supplied UniProt coordinates include omega-N-methylarginine at 544, 950, 1051, 1090, 1101, 1423, 1895, 2016, 2036, and 2074, plus asymmetric dimethylarginine at 1253. Confirm which isoform an antibody detects before interpreting a band; annotated methylation alone does not explain its apparent mass.
Boster reagents
SHANK1 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-SHANK1 Antibody Picoband®
Cat # A05690-2
A05690-2 is a rabbit polyclonal anti-SHANK1 antibody listed for human, mouse, and rat. Its Western blot image shows rat and mouse brain lysates with a band around 220–250 kDa; the supplied evidence does not show a human sample.
Which to pick: A05690-2 is the only listed option. Its WB image uses rat and mouse brain lysates at 0.5 μg/mL. For human samples, reactivity is listed, but no human WB example is supplied.
Source: BosterBio SHANK1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y566.
- Human Protein Atlas. SHANK1 tissue expression.
- PMC6795689 — target-verified WB comparison
- PMC2871978 — target-verified WB comparison
- PMC2745956 — target-verified WB comparison