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- Table of Contents
CDCA8 / Borealin · Western blot design guide
Design a Western Blot for CDCA8
Source-linked CDCA8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CDCA8 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
CDCA8 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 | ~31.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CDCA8 Western Blot Protocol Options
The A06612 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat kidney lysate (catalog A06612) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A06612 · (A) 1 and (B) 2 μg/mL (catalog A06612) |
| 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 CDCA8 Western Blot Band Size?
Borealin has a predicted 31.3 kDa monomer; possible oligomerization and phosphorylation may affect bands, but their effects on migration are unproven.
| Band near 31.3 kDa | Consistent with the predicted Borealin monomer, pending identity controls |
| Band near twice the monomer size | May reflect a homodimer that persists during sample preparation |
| Several bands above the monomer | May reflect persistent Borealin homooligomers |
| Doublet near the monomer | Could reflect phosphorylation states if they resolve; identity requires confirmation |
💡Expected CDCA8 appearanceUniProt predicts a 31.3 kDa Borealin monomer; no empirical band size is supplied, so verify a band near that size with appropriate antibody and CDCA8 depletion controls.
| Predicted monomer mass | Provides a 31.3 kDa reference, not a measured migration position |
| Possible homodimer formation | Could produce a band near twice the monomer size if the dimer persists |
| Possible homooligomer formation | Could produce higher bands if oligomers persist |
| TTK phosphorylation at Thr88 and Thr94 | May alter apparent mobility; a visible shift is not established |
| AURKB phosphorylation at Ser165 | May alter apparent mobility; a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Persistent Borealin homodimers or homooligomers are possible | Compare denaturing conditions and confirm the band by CDCA8 depletion |
| Band lower than expected | Fragment identity is unestablished; UniProt lists no signal peptide or propeptide | Use protease inhibitors and check whether CDCA8 depletion removes the band |
| Multiple bands | Persistent oligomers or resolvable phosphorylation states are possible | Compare sample preparation conditions and confirm each band by CDCA8 depletion |
| Broad smear instead of sharp band | The supplied features do not establish a smear mechanism | Check sample quality and loading, then verify specificity by CDCA8 depletion |
| Weak or no signal | Borealin may be unevenly represented across the sampled cell-cycle states | Check loading and antibody conditions, and compare a mitotic cell sample |
| Fragments below expected size | Sample proteolysis is possible; no physiological cleavage feature is listed | Prepare fresh lysate with protease inhibitors and confirm fragment identity by CDCA8 depletion |
Experiment evidence
Western blot images showing CDCA8 band
A06612 · CDCA8
A06612-2 · CDCA8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CDCA8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CDCA8 in Western blot, you can use appendix tissue, which HPA scores High.
Positive control: Appendix (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: HPA reports no detection in adipose tissue, providing a tissue negative control alongside appendix.
HPA tissue expression evidence for CDCA8
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 |
|---|---|---|---|---|
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | endocrine cells | High | Protein (IHC) | HPA → |
| Skin | fibrohistiocytic cells | High | 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 → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDCA8 Western Blot Tips
Deeper troubleshooting and optimisation questions for CDCA8, answered from its protein features.
How should CDCA8 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 CDCA8 isoforms explain multiple bands?
Isoforms · The supplied entry lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Which phosphorylation sites matter when interpreting CDCA8 bands?
PTM · UniProt coordinates list T88, T94, T169 and T230 phosphorylated by TTK; S165 by AURKB; and T106, S110, T189, T204, S219, S224, S238 and S244 without a specified enzyme. Check the coordinate convention before comparing these sites with antibody or paper numbering. Their presence alone does not establish a band shift.
Does this guide establish induction of CDCA8?
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 CDCA8?
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 A06612 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 CDCA8 bands be quantified across samples?
Quantitation · Quantify the same band region and sample fraction consistently. Record cell-cycle context because CDCA8 is associated with mitosis, and avoid treating a shifted or additional band as a specific phosphorylation state without further evidence.
Where should CDCA8 migrate relative to its predicted mass?
Interpretation · CDCA8 has a predicted mass of 31.3 kDa. No observed band position is supplied, so use 31.3 kDa as a reference, not a confirmed migration position. Its listed phosphorylation sites do not, by themselves, establish a visible shift.
The entry says CDCA8 may form homodimers and homooligomers. Consider that possibility when investigating higher bands, but do not assign a band to an oligomer from this feature alone.
Yes. CDCA8 is linked to mitosis and localizes to chromosome arms and inner centromeres from prophase through metaphase. Compare samples with similar cell-cycle composition when interpreting differences in signal; the supplied features do not establish an induction condition.
The entry places CDCA8 in the nucleus, nucleolus, cytoplasm, spindle and centromere. Keep the fractionation method consistent across samples, and consider both nuclear and cytoplasmic material when a selected fraction gives little signal.
The entry says CDCA8 may form homodimers and homooligomers. Consider that possibility when investigating higher bands, but do not assign a band to an oligomer from this feature alone.
Yes. CDCA8 is linked to mitosis and localizes to chromosome arms and inner centromeres from prophase through metaphase. Compare samples with similar cell-cycle composition when interpreting differences in signal; the supplied features do not establish an induction condition.
The entry places CDCA8 in the nucleus, nucleolus, cytoplasm, spindle and centromere. Keep the fractionation method consistent across samples, and consider both nuclear and cytoplasmic material when a selected fraction gives little signal.
Boster reagents
CDCA8 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-Borealin CDCA8 Antibody
Cat # A06612
Real WB data
Anti-CDCA8 Antibody Picoband®
Cat # A06612-2
Two the supplier anti-CDCA8 antibodies have Western blot images: A06612 in rat kidney lysate and A06612-2 in human cell lysates. The captions document those tested samples, but do not establish performance across every listed reactive species.
Which to pick: For rat kidney lysate, A06612 has an image at 1 and 2 μg/mL. For human cell lysates, A06612-2 shows K562, HEL, A431, and A549 at 0.5 μg/mL. A06612 lists mouse reactivity, but no mouse WB example is supplied.
Source: BosterBio CDCA8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q53HL2.
- Human Protein Atlas. CDCA8 tissue expression.
- PMC11732628 — target-verified WB comparison
- PMC9097394 — target-verified WB comparison
- PMC9066215 — target-verified WB comparison