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CACNA1H / Voltage-dependent T-type calcium channel subunit alpha-1H · Western blot design guide
Design a Western Blot for CACNA1H
Source-linked CACNA1H Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CACNA1H 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
CACNA1H 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 | ~259.2 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Testis (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 | |
| Caveat | Blocking peptide control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CACNA1H Western Blot Protocol Options
The A01406-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 293 cell lysate (catalog A01406-1) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Western blot analysis of CACNA1H in 293 cell lysate with CACNA1H antibody at 1 μg/ml in (A) the absence and (B) the presence of blocking peptide (catalog A01406-1) |
| Primary antibody | A01406-1 · 1 μg/ml (catalog A01406-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected CACNA1H Western Blot Band Size?
CACNA1H is predicted at 259.2 kDa; glycosylation and isoforms could affect migration, but no empirical band or visible size effect is established.
| Band near 259.2 kDa | Consistent with the predicted full-length CACNA1H size; confirm identity with a blocking peptide or other specificity control. |
| Band above 259.2 kDa | N-linked glycosylation could affect migration if the annotated sites are occupied; the size effect is unmeasured. |
| Several high-mass bands | Isoforms 1 and 2 could contribute if they migrate differently; distinct bands are not established. |
| Broad high-mass band | Could reflect variable N-linked glycosylation; site annotations alone do not establish a smear. |
| Weak band in soluble lysate | CACNA1H is a multi-pass cell-membrane protein and may be underrepresented in the soluble fraction. |
💡Expected CACNA1H appearanceUniProt predicts 259.2 kDa, but no empirical band size is supplied; use blocking peptide and membrane-enriched samples to assess the identity of any band near that size.
| Predicted molecular mass | Full-length sequence has a calculated mass of 259.2 kDa. |
| N-linked glycosylation site Asn192 | Occupancy could alter apparent migration; its contribution is unknown. |
| N-linked glycosylation site Asn271 | Occupancy could alter apparent migration; its contribution is unknown. |
| N-linked glycosylation site Asn1466 | Occupancy could alter apparent migration; its contribution is unknown. |
| Splice isoforms 1 and 2 | Their apparent sizes could differ, but their relative sizes and separation are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Multi-pass membrane CACNA1H may be poorly recovered in the soluble lysate. | Check membrane-enriched material and verify extraction of membrane proteins. |
| Band higher than expected | Occupancy of annotated N-linked glycosylation sites could affect migration. | Compare treated and untreated samples and confirm identity with blocking peptide. |
| Band lower than expected | A smaller splice isoform or an unrelated band is possible; neither is established by band size alone. | Use blocking peptide and an independent antibody to check identity. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unconfirmed. | Compare glycosidase-treated and untreated samples and check sample quality. |
| Multiple bands | Isoforms 1 and 2 could differ in migration, or some bands could be unrelated. | Test blocking-peptide competition and compare with an independent antibody. |
| Weak or no signal | Recovery of this membrane protein may be low. | Check membrane-protein extraction and sample loading. |
Experiment evidence
Western blot images showing CACNA1H band
A01406-1 · CACNA1H
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CACNA1H Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CACNA1H in Western blot, you can use testis lysate.
Positive control: Testis (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: As a multi-pass membrane protein, CACNA1H may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for CACNA1H
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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (ciliary rootlets) | Medium | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (ciliary rootlets) | 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 → |
| Pancreas | exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CACNA1H Western Blot Tips
Deeper troubleshooting and optimisation questions for CACNA1H, answered from its protein features.
How should CACNA1H 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.
Can the two CACNA1H isoforms be distinguished on a blot?
Isoforms · Isoform 2 replaces STFPSPE at UniProt positions 1587–1593 with K. This small sequence change does not establish that the isoforms resolve as separate bands. Check whether the antibody epitope includes this region before interpreting isoform-specific detection.
Could glycosylation affect CACNA1H band interpretation?
PTM · UniProt lists N-linked glycosylation at Asn192, Asn271, and Asn1466, using UniProt coordinates. These sites make glycosylation relevant when assessing band patterns, but their presence alone does not demonstrate a visible shift or account for an observed mass difference.
Does this guide establish induction of CACNA1H?
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 CACNA1H Western blot?
Transfer · CACNA1H is a predicted 259.2 kDa multi-pass membrane protein. Select and validate transfer conditions for a protein of this size; inspect the gel after transfer and confirm signal on the membrane to assess recovery.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01406-1 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 CACNA1H signal be quantified?
Quantitation · CACNA1H localizes to the cell membrane, and interaction with STAC increases its membrane expression. Compare like-for-like samples and normalize to an appropriate loading measure. If comparing membrane fractions, account for differences in membrane recovery before attributing signal changes to CACNA1H abundance.
Should CACNA1H run at its predicted 259.2 kDa?
Interpretation · 259.2 kDa is the predicted mass, and no empirical band position is supplied. Use it as a reference when identifying a high-molecular-weight band; the listed features alone cannot establish an apparent mass or explain a difference.
Consider the listed isoform 2 sequence change and N-linked glycosylation sites when reviewing band patterns, but neither feature assigns an unexpected band on its own. Check antibody specificity and compare bands against the predicted 259.2 kDa mass without treating that value as an established migration position.
UniProt reports that STAC interacts with the N-terminal cytoplasmic domain and increases CACNA1H expression at the cell membrane. A membrane-fraction signal may therefore depend on STAC status; interpret such comparisons alongside the fractionation and loading measurements.
Consider the listed isoform 2 sequence change and N-linked glycosylation sites when reviewing band patterns, but neither feature assigns an unexpected band on its own. Check antibody specificity and compare bands against the predicted 259.2 kDa mass without treating that value as an established migration position.
UniProt reports that STAC interacts with the N-terminal cytoplasmic domain and increases CACNA1H expression at the cell membrane. A membrane-fraction signal may therefore depend on STAC status; interpret such comparisons alongside the fractionation and loading measurements.
Boster reagents
CACNA1H 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-CACNA1H Antibody
Cat # A01406-1
A01406-1 is listed for human, mouse, and rat CACNA1H. Its Western blot image uses 293 cell lysate at 1 μg/ml, with and without blocking peptide. No publication evidence or additional specimen testing is supplied.
Which to pick: A01406-1 is the only listed antibody and has a Western blot image. Its documented test uses 293 cell lysate; check suitability for your species and sample type before use.
Source: BosterBio CACNA1H gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95180.
- Human Protein Atlas. CACNA1H tissue expression.
- PMC8225817 — target-verified WB comparison
- PMC9643518 — target-verified WB comparison
- PMC10224624 — target-verified WB comparison