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- Table of Contents
DKC1 / H/ACA ribonucleoprotein complex subunit DKC1 · Western blot design guide
Design a Western Blot for DKC1
Real validated DKC1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DKC1 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
DKC1 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 | ~57.7 kDa | |
| Observed band | ~58 kDa | |
| Gel | 10% (catalog A01535-2) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DKC1 Western Blot Protocols
The A01535-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human U251, human A431 (catalog A01535-2) |
| Gel % | 10% (catalog A01535-2) |
| Load | 30 ug; reducing conditions (catalog A01535-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01535-2) |
| Membrane | nitrocellulose membrane (catalog A01535-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01535-2) |
| Primary antibody | A01535-2 · 0.5 μg/mL (catalog A01535-2) |
| Primary incubation | overnight at 4°C (catalog A01535-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01535-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01535-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01535-2) |
| Detection | ECL (catalog A01535-2) |
Section 2
What Is the Expected DKC1 Western Blot Band Size?
DKC1 is predicted at 57.7 kDa and observed at ~58 kDa; the small difference has no established cause in the supplied evidence.
| Band at ~58 kDa | Matches the reported DKC1 band and its 57.7 kDa predicted mass; confirm identity with antibody controls |
| Additional bands near ~58 kDa | Could reflect isoforms 1 and 3 or phosphorylation states; distinct migration is unconfirmed |
| Band displaced from ~58 kDa | Could reflect a different modification state or isoform; the cause requires testing |
| Faint or absent band in a cytoplasmic fraction | Isoform 1 is reported in the nucleolus and Cajal body |
💡Expected DKC1 appearanceDKC1 is predicted at 57.7 kDa and observed at ~58 kDa in reducing whole-cell blots; confirm any assigned band with antibody specificity controls.
| UniProt predicted mass | Places the reference band near 57.7 kDa; the observed band is ~58 kDa |
| Isoforms 1 and 3 | May differ in size, but their masses and relative migration are not supplied |
| Phosphoserine at residues 21, 387, 451, 453, 455, and 485 | May affect mobility; no visible shift is established |
| Phosphothreonine at residue 458 | May affect mobility; no visible shift is established |
| N-acetylalanine at residue 2 | Is a modification with no established visible band shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear DKC1 signal may be diluted in whole-cell lysate | Check nuclear enrichment and a positive-control lysate |
| Band higher than expected | Its identity or modification state is uncertain; phosphorylation is documented | Compare phosphatase-treated material and use an independent antibody |
| Band lower than expected | An isoform difference is possible, but isoform masses are unknown | Check band identity with an independent antibody or isoform-specific evidence |
| Multiple bands | Isoforms 1 and 3 or distinct phosphorylation states are possible | Compare phosphatase-treated material and verify bands with an independent antibody |
| Weak or no signal | Nuclear localization may limit signal in the sampled fraction | Examine a nuclear fraction and verify sample loading |
Experiment evidence
Western blot images showing DKC1 band
A01535-2 · DKC1
M01535-1 · DKC1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DKC1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for DKC1 in Western blot, you can use adrenal gland lysate, which has high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for DKC1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | non-germinal center cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Medium | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced DKC1 Western Blot Tips
Deeper troubleshooting and optimisation questions for DKC1, answered from its protein features.
How should DKC1 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 DKC1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 3. In isoform 3, canonical residues 420–514 are replaced by one arginine, so a substantially smaller product is possible. Confirm antibody recognition of the retained region before assigning a smaller band to isoform 3.
Choose an antibody whose epitope lies in the sequence retained by both isoforms, before canonical residue 420, and verify that it detects each isoform. An antibody against canonical residues 420–514 would miss that region in isoform 3.
Choose an antibody whose epitope lies in the sequence retained by both isoforms, before canonical residue 420, and verify that it detects each isoform. An antibody against canonical residues 420–514 would miss that region in isoform 3.
Which DKC1 phosphorylation sites matter when interpreting bands?
PTM · In UniProt canonical numbering, phosphoserines occur at 21, 387, 451, 453, 455, 485, 494, and 513; phosphothreonine occurs at 458. Sites 451–513 lie in the region replaced in isoform 3. These annotations alone do not predict a visible band shift.
Yes. UniProt lists N-acetylalanine at canonical position 2. This annotation alone does not establish a separate Western blot band.
Yes. UniProt lists N-acetylalanine at canonical position 2. This annotation alone does not establish a separate Western blot band.
Does this guide establish induction of DKC1?
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.
How should transfer be checked for DKC1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01535-2 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 should DKC1 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why is the DKC1 band near 58 kDa?
Interpretation · Canonical DKC1 has a predicted mass of 57.7 kDa, consistent with the supplied observed band near 58 kDa. Its listed modifications do not establish a visible shift or explain any difference between predicted and apparent mass.
Isoform 3 is one possibility because it replaces canonical residues 420–514 with one arginine. Check whether the antibody recognizes the shared sequence and validate isoform identity before assigning the band. The supplied features do not identify every possible cause of an unexpected band.
Isoform 3 is one possibility because it replaces canonical residues 420–514 with one arginine. Check whether the antibody recognizes the shared sequence and validate isoform identity before assigning the band. The supplied features do not identify every possible cause of an unexpected band.
Boster reagents
DKC1 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-DKC1 Antibody Picoband®
Cat # A01535-2
Real WB data
Anti-DKC1 Rabbit Monoclonal Antibody
Cat # M01535-1
Two the supplier anti-DKC1 antibodies list human, mouse, and rat reactivity and have Western blot images. A01535-2 shows a band near 58 kDa in named human cell and rodent testis lysates; M01535-1 shows SW480 lysate. No orthogonal validation is supplied.
Which to pick: Choose A01535-2 for the documented human cell or rodent testis blot contexts and reported conditions. M01535-1 has a WB image from SW480 lysate, but its supplied caption gives fewer experimental details.
Source: BosterBio DKC1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60832.
- Human Protein Atlas. DKC1 tissue expression.
- PMC10307871 — target-verified WB comparison
- PMC7054532 — target-verified WB comparison
- PMC8056518 — target-verified WB comparison