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- Table of Contents
CDC5L / Cell division cycle 5-like protein · Western blot design guide
Design a Western Blot for CDC5L
Source-linked CDC5L Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CDC5L 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
CDC5L 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 | ~92.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 CDC5L Western Blot Protocol Options
The M03797 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M03797) |
| Gel % | 8–10% (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 | M03797; use the WB datasheet starting dilution (standard starting point) |
| 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 CDC5L Western Blot Band Size?
CDC5L's predicted monomer mass is 92.3 kDa; homodimerization and phosphorylation could affect its pattern, but no altered migration or empirical band size is established.
| Band near 92.3 kDa | Consistent with the predicted CDC5L monomer; identity requires validation |
| Possible band near 185 kDa | Could reflect a retained homodimer if sample conditions preserve it |
| Close bands near 92.3 kDa | Could reflect different phosphorylation states; their migration is unverified |
| Weak band in a nuclear-depleted fraction | Consistent with CDC5L's nuclear localization |
💡Expected CDC5L appearanceCDC5L has a predicted monomer mass of 92.3 kDa, but no empirical band size is supplied; confirm any candidate band with ordinary identity controls.
| Predicted monomer mass | Provides a 92.3 kDa reference, not a measured band position |
| Homodimer formation | Could produce a band near twice the monomer mass if the dimer survives sample preparation |
| Phosphorylation at Thr227 | May affect migration, but no visible shift is established |
| Phosphorylation at Ser303 | May affect migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear CDC5L may be poorly recovered during extraction | Check nuclear extraction and compare with a nuclear fraction |
| Band higher than expected | A CDC5L homodimer may persist under the preparation conditions | Compare fully denatured and reducing preparations |
| Band lower than expected | The supplied features do not establish a smaller CDC5L form | Verify identity with an independent antibody or CDC5L depletion |
| Multiple bands | Different phosphorylation states are possible, but their migration is unverified | Compare phosphatase-treated material and validate each band |
| Weak or no signal | Nuclear protein recovery may be low | Check extraction with a nuclear marker and include a positive lysate control |
Experiment evidence
Western blot images showing CDC5L band
M03797 · CDC5L
PA2123 · CDC5L
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CDC5L Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CDC5L in Western blot, you can use adrenal gland tissue, which HPA scores as high for CDC5L.
Positive control: Adrenal gland (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: Adipose tissue is a candidate negative control, but confirm its lack of signal by Western blot.
HPA tissue expression evidence for CDC5L
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 | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial 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 → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced CDC5L Western Blot Tips
Deeper troubleshooting and optimisation questions for CDC5L, answered from its protein features.
How should CDC5L 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 CDC5L isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning multiple bands to CDC5L isoforms.
Which CDC5L phosphorylation sites matter when assessing band patterns?
PTM · UniProt lists phosphothreonine at 227, 377, 385, 396, 404, 411, 415, 424, 430, 438 and 442, and phosphoserine at 303, 358, 417 and 437. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish a visible band shift.
Does this guide establish induction of CDC5L?
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 CDC5L?
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 M03797 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 CDC5L be quantified on a blot?
Quantitation · Quantify a consistently identified band near the 92.3 kDa predicted mass, using the same band definition across samples. If multiple bands appear, the supplied features cannot establish which represents CDC5L; verify band identity before combining their signals.
Where should CDC5L migrate relative to its predicted mass?
Interpretation · The predicted mass is 92.3 kDa. No observed band position is supplied, so use 92.3 kDa as a reference rather than an expected apparent mass. The listed modifications alone do not establish a visible shift or explain any difference.
CDC5L is listed as a homodimer and has 15 phosphorylated residues, but neither feature identifies an unexpected band. The record lists no glycosylation sites or alternative sequence. Verify band identity before attributing a lower or higher band to cleavage, modification or oligomerization.
CDC5L is listed in the nucleus and as a component of spliceosome complexes. A nuclear fraction is therefore relevant when checking detection or comparing fractionated samples. The supplied record also includes a cytoplasm keyword, so fraction identity should be verified rather than inferred from a band alone.
CDC5L is listed as a homodimer and has 15 phosphorylated residues, but neither feature identifies an unexpected band. The record lists no glycosylation sites or alternative sequence. Verify band identity before attributing a lower or higher band to cleavage, modification or oligomerization.
CDC5L is listed in the nucleus and as a component of spliceosome complexes. A nuclear fraction is therefore relevant when checking detection or comparing fractionated samples. The supplied record also includes a cytoplasm keyword, so fraction identity should be verified rather than inferred from a band alone.
Boster reagents
CDC5L 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-CDC5 CDC5L Rabbit Monoclonal Antibody
Cat # M03797
Real WB data
Anti-CDC5L Antibody Picoband®
Cat # PA2123
Two the supplier anti-CDC5L antibodies have Western blot images: M03797 in HeLa lysate and PA2123 in HEK293, THP-1, and rat C6 lysates. PA2123 shows a band near 100 kDa versus an expected 92 kDa. The supplied evidence does not establish orthogonal validation.
Which to pick: For mouse samples, M03797 is the listed option with mouse reactivity, though its supplied blot uses human HeLa lysate. For human or rat samples, PA2123 provides documented blot conditions and tested lysates; M03797 also lists human and rat reactivity.
Source: BosterBio CDC5L gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99459.
- Human Protein Atlas. CDC5L tissue expression.
- PMC6930429 — target-verified WB comparison
- PMC12747968 — target-verified WB comparison
- PMC13582790 — target-verified WB comparison