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- Table of Contents
HAND2 / Heart- and neural crest derivatives-expressed protein 2 · Western blot design guide
Design a Western Blot for HAND2
Real validated HAND2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HAND2 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
HAND2 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 | ~23.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Placenta (IHC candidate; verify WB) +4 more | |
| Negative control | Lymph node (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated HAND2 Western Blot Protocols
The A04652 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04652) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04652) |
| Primary antibody | A04652 · 1:1000 (catalog A04652) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04652) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04652) |
Section 2
What Is the Expected HAND2 Western Blot Band Size?
HAND2 has a predicted 23.7 kDa monomer; isoforms and homodimerization could affect patterns, but no empirical migration or distinct isoform bands are demonstrated.
| Band near 23.7 kDa | Consistent with predicted HAND2 monomer size; confirm identity with a specific control |
| Several bands of different sizes | Isoforms 1 and 2 could contribute if they migrate differently; their sizes are unknown |
| Band near twice the monomer size under native conditions | Could reflect a retained HAND2 homodimer; identity needs confirmation |
| Little or no band in a cytoplasmic fraction | HAND2 is annotated as nuclear |
💡Expected HAND2 appearanceUniProt predicts a 23.7 kDa HAND2 monomer, but no empirical band size is supplied; confirm any candidate band with a positive control and HAND2 depletion.
| Predicted HAND2 monomer mass | Provides a 23.7 kDa reference, not a measured migration position |
| Splice isoform 1 | Could migrate differently from isoform 2; its individual size is unknown |
| Splice isoform 2 | Could migrate differently from isoform 1; its individual size is unknown |
| HAND2 homodimer formation | Could yield a band near twice the monomer size if the complex survives electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HAND2 may be poorly represented in the preparation | Check a nuclear fraction and a HAND2-positive control |
| Band higher than expected | A HAND2 complex may persist under native conditions | Compare denaturing conditions and confirm identity by HAND2 depletion |
| Band lower than expected | The band may represent an isoform or an unrelated protein; no isoform size is supplied | Compare isoform-specific expression and HAND2 depletion |
| Multiple bands | Isoforms 1 and 2 may differ in migration, but distinct bands are unproven | Test each band with HAND2 depletion or isoform-specific controls |
| Weak or no signal | Nuclear localization may limit HAND2 recovered from whole-cell lysate | Enrich the nuclear fraction and include a positive control |
| Fragments below expected size | Fragment identity is not established by the supplied HAND2 features | Use fresh lysate with protease inhibitors and test band identity by HAND2 depletion |
Experiment evidence
Western blot images showing HAND2 band
A04652 · HAND2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HAND2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HAND2 in Western blot, you can use placenta, the highest-scoring HPA tissue.
Positive control: Placenta (IHC candidate; verify WB)
Negative control: Lymph node (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because HAND2 is nuclear, a nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for HAND2
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 |
|---|---|---|---|---|
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Thyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced HAND2 Western Blot Tips
Deeper troubleshooting and optimisation questions for HAND2, answered from its protein features.
How should HAND2 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 HAND2 isoforms produce different bands?
Isoforms · Yes. UniProt lists two isoforms. Relative to the canonical sequence, isoform 2 lacks residues 21–26 and 71–101, so it has a shorter sequence. These features do not establish where either isoform migrates on a blot.
Check the antibody epitope against the two deleted regions. An antibody recognizing canonical residues 21–26 or 71–101 may not detect isoform 2. Use UniProt coordinates when comparing epitope positions; antibody documentation may use a different numbering convention.
Check the antibody epitope against the two deleted regions. An antibody recognizing canonical residues 21–26 or 71–101 may not detect isoform 2. Use UniProt coordinates when comparing epitope positions; antibody documentation may use a different numbering convention.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of HAND2?
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 HAND2 Western blot?
Transfer · HAND2 is predicted to be 23.7 kDa. Choose transfer conditions suitable for a protein of that size, then check transfer and membrane retention with a stain or size marker. The supplied features do not specify a membrane, transfer time or voltage.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04652 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 HAND2 bands be quantified?
Quantitation · HAND2 is annotated as nuclear and has two isoforms. Compare like samples and fractions, and define whether quantification covers one band or both putative isoforms. Confirm band identity before combining signals; sequence differences alone do not establish either band’s position.
Should HAND2 migrate at its predicted 23.7 kDa?
Interpretation · The canonical 217-residue sequence has a predicted mass of 23.7 kDa, but no observed band position is supplied. The listed features do not establish a visible shift or explain a difference between predicted and apparent mass.
First consider the shorter isoform 2, which lacks canonical residues 21–26 and 71–101. HAND2 also forms homo- and heterodimers with other bHLH proteins, but that annotation alone does not show that a dimer survives blot preparation. The supplied features cannot assign an unexpected band’s identity.
UniProt places HAND2 in the nucleus. A nuclear fraction is therefore a reasonable sample to examine, alongside an appropriate fraction control. Its annotated location does not establish how abundant HAND2 will be in a particular sample.
First consider the shorter isoform 2, which lacks canonical residues 21–26 and 71–101. HAND2 also forms homo- and heterodimers with other bHLH proteins, but that annotation alone does not show that a dimer survives blot preparation. The supplied features cannot assign an unexpected band’s identity.
UniProt places HAND2 in the nucleus. A nuclear fraction is therefore a reasonable sample to examine, alongside an appropriate fraction control. Its annotated location does not establish how abundant HAND2 will be in a particular sample.
Boster reagents
HAND2 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-HAND2 Antibody
Cat # A04652
the supplier A04652 is an anti-HAND2 antibody listed for human, mouse, and rat. Its Western blot image uses extracts from various cell lines at 1:1000 dilution, with 25 µg per lane. The caption does not identify the cell lines or their species.
Which to pick: A04652 is the only listed option and has a Western blot image. Check its reported reactivity against your sample species; the image caption does not establish which species were tested.
Source: BosterBio HAND2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P61296.
- Human Protein Atlas. HAND2 tissue expression.
- PMC9308366 — target-verified WB comparison
- PMC10845658 — target-verified WB comparison
- PMC3046314 — target-verified WB comparison