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- Table of Contents
CDK3 / Cyclin-dependent kinase 3 · Western blot design guide
Design a Western Blot for CDK3
Source-linked CDK3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CDK3 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
CDK3 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 | ~35 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bronchus (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CDK3 Western Blot Protocol Options
The A08097 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HEK293T, Raw264.7, PC12 (catalog A08097) |
| 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 | A08097; 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 CDK3 Western Blot Band Size?
CDK3 has a predicted mass of 35 kDa; no empirical band is supplied, and the listed features do not demonstrate a migration shift.
| Single sharp band near 35 kDa | Consistent with the predicted size of CDK3 |
| Band near 35 kDa in whole-cell lysate | Candidate full-length CDK3; identity needs validation |
| No band near 35 kDa | CDK3 is not detected under these conditions |
| Additional bands away from 35 kDa | Their identity is not explained by the supplied CDK3 features |
💡Expected CDK3 appearanceUniProt predicts CDK3 at 35 kDa; no empirical band size is supplied, so a band near 35 kDa is a candidate that needs ordinary band-identity controls.
| UniProt predicted mass | Places the expected full-length band near 35 kDa |
| Calculated mass of 35,046 Da | Supports an approximately 35 kDa size estimate |
| Full-length predicted mass | Provides a reference for assessing other band positions |
| Sequence-based mass estimate | Does not establish the measured migration position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CDK3 may be below detection under these conditions | Check loading and assay sensitivity with a positive control |
| Band higher than expected | Identity or migration difference is unestablished | Compare with a positive control and verify antibody specificity |
| Band lower than expected | Identity or migration difference is unestablished | Check specificity with CDK3 depletion or an independent antibody |
| Multiple bands | The supplied features do not assign additional CDK3 species | Identify the CDK3-dependent band using depletion and a positive control |
| Weak or no signal | CDK3 may be scarce or assay sensitivity insufficient | Check loading, antibody performance, and a positive control |
| Fragments below expected size | The supplied features do not establish CDK3 fragments | Check sample integrity and confirm band identity with an independent antibody |
Experiment evidence
Western blot images showing CDK3 band
A08097 · CDK3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CDK3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CDK3 in Western blot, you can use bronchus tissue, which HPA rates High.
Positive control: Bronchus (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 CDK3 as not detected in adipose tissue, making a tissue negative control feasible.
HPA tissue expression evidence for CDK3
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 |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Liver | hepatocytes | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDK3 Western Blot Tips
Deeper troubleshooting and optimisation questions for CDK3, answered from its protein features.
Where should the main CDK3 band appear?
Band shift · The supplied sequence is 305 residues with a predicted mass of 35 kDa, so use 35 kDa as the starting position. No observed band position is supplied; an apparent shift cannot be inferred from the listed features.
Should CDK3 isoforms produce multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not identify an isoform that would explain multiple bands.
Could a listed modification explain a shifted CDK3 band?
PTM · No modified residues or glycosylation sites are listed. These features provide no specific modification to assign to a shifted band; they also do not establish why a band differs from the predicted 35 kDa.
Does this guide establish induction of CDK3?
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 CDK3 Western blot?
Transfer · Plan transfer around the predicted 35 kDa protein and check the 35 kDa marker region on the membrane. The supplied features do not specify a transfer method or establish which settings will recover CDK3 best.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A08097 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 CDK3 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.
Does cyclin C binding predict increased CDK3 band intensity?
Interpretation · Binding to CCNC/cyclin-C promotes RB1 phosphorylation, but the supplied features do not say that binding increases CDK3 abundance. Quantify the CDK3 band separately from any assay of pathway activity.
Compare them with the predicted 35 kDa position and verify their identity independently. CDK3 interacts with ATF1 and, by similarity, CABLES1 and CABLES2, but these interactions alone do not identify an unexpected band or show that a complex survives Western blot preparation.
Compare them with the predicted 35 kDa position and verify their identity independently. CDK3 interacts with ATF1 and, by similarity, CABLES1 and CABLES2, but these interactions alone do not identify an unexpected band or show that a complex survives Western blot preparation.
Boster reagents
CDK3 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-Cyclin-dependent kinase 3 CDK3 Antibody
Cat # A08097
The catalog reports one anti-CDK3 antibody, A08097, with reported human, mouse, and rat reactivity. Its WB image shows HEK293T, Raw264.7, and PC12 whole cell lysates at 1:500 dilution. No publication evidence was supplied.
Which to pick: A08097 is the only listed option. Its WB image includes the three named cell lysates; choose it based on your sample and the reported reactivity, while recognizing that those examples do not establish universal validation.
Source: BosterBio CDK3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q00526.
- Human Protein Atlas. CDK3 tissue expression.
- PMC6532885 — target-verified WB comparison
- PMC9404285 — target-verified WB comparison
- PMC4808043 — target-verified WB comparison