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- Table of Contents
CDKN3 / Cyclin-dependent kinase inhibitor 3 · Western blot design guide
Design a Western Blot for CDKN3
Real validated CDKN3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CDKN3 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
CDKN3 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 | ~23.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated CDKN3 Western Blot Protocols
The A05157-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF7, (catalog A05157-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05157-1) |
| Primary antibody | A05157-1 · 1:400 (catalog A05157-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05157-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05157-1) |
Section 2
What Is the Expected CDKN3 Western Blot Band Size?
CDKN3 is predicted at 23.8 kDa; isoforms may affect migration, but no empirical band size or distinct isoform pattern is established.
| Single band near 23.8 kDa | Consistent with the UniProt predicted mass; confirm band identity with controls |
| Two discrete bands | Could reflect isoforms 1 and 2 if they migrate differently |
| One dominant band with a faint second band | Could reflect unequal detection of isoforms 1 and 2 if their migration differs |
| Band away from 23.8 kDa | Its identity and the reason for its apparent size remain unestablished |
💡Expected CDKN3 appearanceUniProt predicts 23.8 kDa for CDKN3, but no empirical band size is supplied; isoforms 1 and 2 may differ in migration, so confirm any candidate band with identity controls.
| UniProt predicted mass | Sets a 23.8 kDa sequence-based reference, not a measured band position |
| Isoform 1 | May contribute a band whose individual mass is not supplied |
| Isoform 2 | May migrate differently from isoform 1; its individual mass is not supplied |
| Alternative splicing | Can change sequence length, though distinct bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Identity or apparent migration is uncertain | Compare with a CDKN3 knockdown control |
| Band lower than expected | Isoform migration is possible but unverified | Compare with a CDKN3 knockdown control |
| Multiple bands | Isoforms 1 and 2 could migrate differently | Check which bands decrease after CDKN3 knockdown |
| Weak or no signal | CDKN3 abundance in the sample may be low | Check lysate loading and use a CDKN3-positive control |
| Fragments below expected size | Fragment identity is unestablished | Compare fresh lysate with a CDKN3 knockdown control |
Experiment evidence
Western blot images showing CDKN3 band
A05157-1 · CDKN3
A05157 · CDKN3
A05157 · CDKN3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CDKN3 Western blot
🧪For positive controls for CDKN3 in Western blot, you can use a tissue or cell line once HPA data identify a positive sample.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA data are unavailable, so neither a positive sample nor a naturally negative tissue can be verified.
HPA tissue expression evidence for CDKN3
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced CDKN3 Western Blot Tips
Deeper troubleshooting and optimisation questions for CDKN3, answered from its protein features.
How should CDKN3 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 CDKN3 isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 11–50 of the canonical sequence, so it has a shorter sequence. Compare candidate bands with isoform-specific sequence information; the feature alone does not establish their apparent positions.
Do listed modifications explain a shifted CDKN3 band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. These records therefore provide no modification-based explanation for a shift, but they cannot establish the cause of an observed band.
Does this guide establish induction of CDKN3?
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 CDKN3 Western blot?
Transfer · The supplied features give a predicted mass of 23.8 kDa but do not specify a transfer method. Choose and verify transfer conditions that retain a protein near this size, checking transfer efficiency before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05157-1 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.
How should CDKN3 bands be quantified across samples?
Quantitation · Quantify the same validated band consistently across samples. Because two isoforms are listed, decide whether the assay measures one isoform or both before combining band signals; the supplied features do not establish which isoform an antibody detects.
Where should CDKN3 migrate on a Western blot?
Interpretation · The predicted mass is 23.8 kDa for the 212-residue canonical sequence. No observed band position is supplied, so use 23.8 kDa as a starting reference, not a guaranteed apparent mass.
Consider isoform 2, which lacks canonical residues 11–50, as one possibility. Band size alone cannot identify it. Check whether the antibody recognizes sequence retained in isoform 2 before assigning the band.
Consider isoform 2, which lacks canonical residues 11–50, as one possibility. Band size alone cannot identify it. Check whether the antibody recognizes sequence retained in isoform 2 before assigning the band.
Boster reagents
CDKN3 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-CDKN3 Antibody
Cat # A05157-1
Real WB data
Anti-CDKN3 Antibody
Cat # A05157
Both listed anti-CDKN3 antibodies report Human and Mouse reactivity and include Western blot images. The supplied captions show MCF7 and SH-SY5Y cell samples; neither documents a Mouse sample.
Which to pick: Choose A05157-1 if the MCF7 blot conditions (1:400 antibody dilution, 25 µg per lane) fit your setup, or A05157 for the SH-SY5Y example at 1:1000. Both have WB images; the supplied captions do not show Mouse samples.
Source: BosterBio CDKN3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16667.
- Human Protein Atlas. CDKN3 tissue expression.
- PMC8977226 — target-verified WB comparison
- PMC7816303 — target-verified WB comparison
- PMC7841180 — target-verified WB comparison