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DYNC1H1 / Cytoplasmic dynein 1 heavy chain 1 · Western blot design guide
Design a Western Blot for DYNC1H1
Source-linked DYNC1H1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DYNC1H1 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
DYNC1H1 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 | ~532.4 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Testis (IHC candidate; verify WB) +4 more | |
| Negative control | Esophagus (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked DYNC1H1 Western Blot Protocol Options
The A03059-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysates from Hela, PC-3 cell line and rat brain tissue lysate (from left to right), (catalog A03059-1) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03059-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-rabbit IgG, 1:10000 (catalog A03059-1) |
| 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 DYNC1H1 Western Blot Band Size?
DYNC1H1 has a predicted monomer mass of 532.4 kDa; homodimerization and documented modifications are possible influences, but their effects on migration are unproven.
| Band near 532.4 kDa | consistent with the predicted DYNC1H1 monomer |
| Band near 1,065 kDa | could reflect a homodimer retained during sample preparation |
| Weak band in soluble lysate | cytoskeletal DYNC1H1 may be underrepresented in the soluble fraction |
| No distinct modification-dependent shift | the listed phosphorylation and acetylation sites do not establish a resolvable size change |
💡Expected DYNC1H1 appearanceThe predicted DYNC1H1 monomer is 532.4 kDa; no empirical band size is supplied, so confirm any detected band with molecular-weight markers and an independent antibody or depletion control.
| UniProt predicted monomer mass | places the expected full-length band near 532.4 kDa |
| Homodimer formation | could yield a band near twice the monomer mass if the complex survives sample preparation |
| Documented phosphoserine and phosphothreonine sites | do not establish a resolvable change in apparent size |
| Documented acetylation sites | do not establish a resolvable change in apparent size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytoskeletal DYNC1H1 may be poorly recovered | check the insoluble fraction and verify extraction |
| Band higher than expected | a homodimer may persist during sample preparation | compare denaturing conditions and confirm identity with an independent antibody |
| Band lower than expected | possible breakdown of the large DYNC1H1 chain | compare antibodies against different regions and use a depletion control |
| Weak or no signal | incomplete recovery of cytoskeletal protein | optimize extraction and check lysate loading |
| Fragments below expected size | possible breakdown of the 4,646-residue protein | prepare lysate with protease inhibitors and compare antibodies against different regions |
| Multiple bands | band identities are unresolved; the supplied record lists no named alternative isoforms | check specificity with an independent antibody or depletion control |
Experiment evidence
Western blot images showing DYNC1H1 band
A03059-1 · DYNC1H1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DYNC1H1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DYNC1H1 in Western blot, you can use testis lysate, which has high HPA expression.
Positive control: Testis (IHC candidate; verify WB)
Negative control: Esophagus (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA identifies testis as positive and esophagus as not detected, making tissue controls feasible.
HPA tissue expression evidence for DYNC1H1
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 | spermatogonia cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | 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 |
|---|---|---|---|---|
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced DYNC1H1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DYNC1H1, answered from its protein features.
Where should the DYNC1H1 band appear relative to its predicted mass?
Band shift · The predicted mass is 532.4 kDa. No observed band position is supplied, so there is no measured difference to explain. The listed modifications alone do not establish a visible shift.
Could annotated isoforms explain multiple DYNC1H1 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no annotated isoform explanation for multiple bands.
Which DYNC1H1 phosphorylation sites could matter when interpreting bands?
PTM · UniProt lists phosphoserine at positions 70, 1230, 4162, and 4368, and phosphothreonine at 4366. These are UniProt coordinates; antibody or paper numbering may differ. Site annotation alone does not establish a visible band shift.
UniProt lists N-acetylserine at position 2 and N6-acetyllysine at positions 1125, 3480, and 4283. These are UniProt coordinates; check the numbering convention when comparing antibody or paper labels. These annotations do not by themselves explain an apparent mass difference.
UniProt lists N-acetylserine at position 2 and N6-acetyllysine at positions 1125, 3480, and 4283. These are UniProt coordinates; check the numbering convention when comparing antibody or paper labels. These annotations do not by themselves explain an apparent mass difference.
Does this guide establish induction of DYNC1H1?
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 DYNC1H1 Western blot?
Transfer · DYNC1H1 is a 532.4 kDa protein. Use transfer conditions validated for proteins of this size, and check whether the high molecular weight protein transferred before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03059-1 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.
What should be checked when quantifying DYNC1H1 bands?
Quantitation · DYNC1H1 is a 532.4 kDa cytoplasmic cytoskeletal protein. Confirm that the band being measured is DYNC1H1 and that transfer is adequate at its size. The supplied modification sites do not establish which, if any, separate bands should be combined for total protein quantitation.
How should unexpected lower DYNC1H1 bands be interpreted?
Interpretation · The supplied features list no propeptide, signal peptide, or alternative sequence that explains a smaller product. Compare unexpected bands with the 532.4 kDa prediction, and verify their identity before assigning them to DYNC1H1.
UniProt describes DYNC1H1 as a homodimer within the cytoplasmic dynein 1 complex. That annotation alone does not establish that a higher band is a dimer; interpret it in light of the sample preparation and confirm its identity.
UniProt describes DYNC1H1 as a homodimer within the cytoplasmic dynein 1 complex. That annotation alone does not establish that a higher band is a dimer; interpret it in light of the sample preparation and confirm its identity.
Boster reagents
DYNC1H1 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-DYNC1H1 Antibody (C-term)
Cat # A03059-1
A03059-1 is an anti-DYNC1H1 C-terminal antibody listed for human and rat. Its WB image shows HeLa and PC-3 cell lysates and rat brain tissue lysate at 35 µg per lane, with the primary antibody diluted 1:1000. Evidence is limited to these reported samples.
Which to pick: A03059-1 is the only listed antibody and has a WB image. It is the practical choice for the shown human cell lines or rat brain tissue; the supplied evidence does not establish performance in other samples.
Source: BosterBio DYNC1H1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14204.
- Human Protein Atlas. DYNC1H1 tissue expression.
- PMC13237520 — target-verified WB comparison
- PMC12622415 — target-verified WB comparison
- PMC6598784 — target-verified WB comparison