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- Table of Contents
HAND1 / Heart- and neural crest derivatives-expressed protein 1 · Western blot design guide
Design a Western Blot for HAND1
Real validated HAND1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HAND1 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
HAND1 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 | ~23.6 kDa | |
| Observed band | ~24 kDa | |
| Gel | 12% (catalog A06496-3) | |
| Positive control | Caudate (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated HAND1 Western Blot Protocols
The A06496-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SH-SY5Y (catalog A06496-3) |
| Gel % | 12% (catalog A06496-3) |
| Load | 30 ug; reducing conditions (catalog A06496-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06496-3) |
| Membrane | nitrocellulose membrane (catalog A06496-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06496-3) |
| Primary antibody | A06496-3 · 0.5 μg/mL (catalog A06496-3) |
| Primary incubation | overnight at 4°C (catalog A06496-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06496-3) |
| Secondary incubation | 1.5 hour at RT (catalog A06496-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06496-3) |
| Detection | ECL (catalog A06496-3) |
Section 2
What Is the Expected HAND1 Western Blot Band Size?
HAND1 is predicted at 23.6 kDa and observed at ~24 kDa; the small difference has no established cause.
| Band at ~24 kDa | Matches the empirical HAND1 band and its 23.6 kDa predicted mass |
| Faint ~24 kDa band | May reflect limited recovery of nuclear HAND1 |
| Higher band near twice the monomer size | Could reflect a retained HAND1 homodimer; band identity needs confirmation |
| Close doublet near ~24 kDa | Could reflect different phosphorylation states; their migration is not established |
💡Expected HAND1 appearanceExpect a band at the empirical ~24 kDa, close to HAND1's 23.6 kDa predicted mass; confirm identity with appropriate antibody and sample controls.
| UniProt predicted mass | Places the 215-residue protein near 23.6 kDa; the empirical band is ~24 kDa |
| HAND1 homodimer formation | Could yield a higher band if the complex survives sample preparation; persistence on SDS-PAGE is unproven |
| PLK4 phosphorylation at Thr107 | May alter migration, but a visible shift is not established |
| PLK4 phosphorylation at Ser109 | May alter migration, but a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HAND1 may be poorly recovered | Check nuclear extraction and use a positive control |
| Band higher than expected | A retained HAND1 complex is possible | Compare denaturing conditions and confirm band identity |
| Band lower than expected | Protein degradation is possible; no cleavage feature is listed | Prepare fresh lysate with protease inhibitors and confirm band identity |
| Multiple bands | Phosphorylation states are possible, but their migration is unproven | Compare phosphatase-treated samples and verify specificity |
| Weak or no signal | Limited nuclear HAND1 recovery or low sample abundance | Check loading, nuclear recovery and a positive control |
Experiment evidence
Western blot images showing HAND1 band
A06496-3 · HAND1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HAND1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HAND1 in Western blot, you can use caudate tissue, which has high HPA expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HAND1 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for HAND1
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 |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HAND1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HAND1, answered from its protein features.
Where should the HAND1 band appear?
Band shift · HAND1 has a predicted mass of 23.6 kDa, and the supplied Western blot observation is approximately 24 kDa. Use that region as the starting point for band identification.
Could HAND1 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It does not support assigning additional bands to named HAND1 isoforms.
Could phosphorylation shift the HAND1 band?
PTM · UniProt lists phosphothreonine at position 107 and phosphoserine at position 109, both by PLK4. These modifications may affect migration, but their presence alone does not establish a visible shift or explain a mass difference. The positions use UniProt numbering, which may differ from antibody or paper numbering.
Does this guide establish induction of HAND1?
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 HAND1 Western blot?
Transfer · HAND1 is a 215-residue protein with a predicted mass of 23.6 kDa. Check transfer and membrane retention around 24 kDa using a molecular-weight marker. The supplied features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06496-3 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 HAND1 bands be quantified?
Quantitation · Quantify the approximately 24 kDa band consistently across samples. If comparing nuclear fractions, account for HAND1's reported nucleoplasmic and nucleolar locations; PLK4 phosphorylation is linked to release from the nucleolus through disrupted MDFIC interaction.
Can HAND1 localization change without a new band?
Interpretation · Yes. The supplied annotation links PLK4 phosphorylation to disruption of HAND1's interaction with MDFIC and release from the nucleolus. A change in nucleolar versus nucleoplasmic signal need not produce a resolvable change in band position.
HAND1 can form homodimers and heterodimers with other bHLH proteins, but this annotation does not establish that an unexpected Western blot band is a dimer. Compare it with the approximately 24 kDa band and verify its identity before assigning it to a HAND1 complex.
HAND1 can form homodimers and heterodimers with other bHLH proteins, but this annotation does not establish that an unexpected Western blot band is a dimer. Compare it with the approximately 24 kDa band and verify its identity before assigning it to a HAND1 complex.
Boster reagents
HAND1 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-HAND1 Antibody Picoband®
Cat # A06496-3
A06496-3 is a human-reactive anti-HAND1 antibody with a Western blot image from human SH-SY5Y whole-cell lysate. The caption reports a band near the expected 24 kDa using 0.5 μg/mL primary antibody. Evidence is limited to this reported sample and blot.
Which to pick: Only A06496-3 is listed. Choose it for a human HAND1 Western blot when the SH-SY5Y lysate example is relevant; its caption provides a 24 kDa band and assay conditions. Validation in other specimens or species is not shown.
Source: BosterBio HAND1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O96004.
- Human Protein Atlas. HAND1 tissue expression.
- PMC9898290 — target-verified WB comparison
- PMC10845658 — target-verified WB comparison
- PMC9760445 — target-verified WB comparison