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- Table of Contents
CCNE2 / G1/S-specific cyclin-E2 · Western blot design guide
Design a Western Blot for CCNE2
Real validated CCNE2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CCNE2 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
CCNE2 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 | ~46.8 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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated CCNE2 Western Blot Protocols
The M04536 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 M04536) |
| 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 | M04536; 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 CCNE2 Western Blot Band Size?
CCNE2 is predicted at 46.8 kDa; isoforms and listed modifications could affect migration, but no empirical band or feature-specific shift is established.
| Band near 46.8 kDa | Consistent with the predicted CCNE2 mass; confirm identity with controls |
| Lower band | Could represent the Short isoform; its migration is not supplied |
| Two discrete bands | Could represent Long and Short isoforms if both are expressed and resolved |
| Closely spaced bands | Could reflect modified CCNE2, but the listed modifications do not establish a visible shift |
💡Expected CCNE2 appearanceCCNE2 has a predicted mass of 46.8 kDa, but no empirical band size is supplied; use antibody and band-identity controls to assess any observed band.
| UniProt predicted mass | Places the reference full-length size at 46.8 kDa |
| Long isoform | May differ in size from Short; its individual mass is not supplied |
| Short isoform | May be smaller than Long; its individual mass is not supplied |
| Phosphorylation at Ser21, Ser383 and Thr392 | May alter migration, but a visible shift is not established |
| N6-lactoyllysine at Lys348 | May alter migration, but a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear CCNE2 may be poorly represented in the preparation | Check nuclear recovery and compare with the HeLa lysate used in antibody QC |
| Band higher than expected | The listed modifications do not establish the cause of a higher band | Confirm identity with an independent antibody or CCNE2 depletion |
| Band lower than expected | The Short isoform is a possibility, but its migration is unknown | Check isoform expression and confirm band identity |
| Multiple bands | Long and Short isoforms or modified states are possible | Confirm bands with isoform-aware controls or CCNE2 depletion |
| Weak or no signal | Target abundance or nuclear recovery may be low | Check loading, nuclear recovery and a positive-control lysate |
| Fragments below expected size | Protein breakdown is possible; no cleavage feature is listed | Compare fresh, protease-inhibited lysate and confirm fragment identity |
Experiment evidence
Western blot images showing CCNE2 band
M04536 · CCNE2
M04536-1 · CCNE2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CCNE2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CCNE2 in Western blot, you can use appendix tissue, which shows medium HPA expression.
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: CCNE2 is nuclear, so nuclear-enriched lysate may help if whole-cell signal is weak.
HPA tissue expression evidence for CCNE2
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 | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | Medium | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCNE2 Western Blot Tips
Deeper troubleshooting and optimisation questions for CCNE2, answered from its protein features.
Where should the main CCNE2 band appear?
Band shift · The predicted mass of the 404-residue canonical sequence is 46.8 kDa. No empirical apparent band position is supplied, so use 46.8 kDa as a reference rather than an expected exact migration position.
Could CCNE2 isoforms produce different bands?
Isoforms · UniProt lists Long and Short isoforms. Short lacks residues 167–211 of the canonical sequence, so it may migrate differently. Confirm which isoform the antibody recognizes before assigning bands; the supplied features do not establish either isoform’s apparent band position.
Which CCNE2 modifications matter when interpreting bands?
PTM · The supplied UniProt coordinates list phosphoserine at 21 and 383, N6-lactoyllysine at 348, and phosphothreonine at 392. These coordinates refer to the supplied canonical sequence; numbering in another isoform or an antibody document may differ. Their presence alone does not establish a visible shift.
Not on their own. UniProt lists four modified residues, but supplies no evidence that they cause a visible shift or account for an observed-versus-calculated mass difference. Check isoform recognition and band specificity before assigning the difference to a modification.
No glycosylation sites, signal peptide, or propeptide are listed. Those features therefore provide no basis here for attributing an unexpected band to glycosylation or precursor cleavage. Assess the listed isoforms and modified residues without assuming either produces a visible shift.
Not on their own. UniProt lists four modified residues, but supplies no evidence that they cause a visible shift or account for an observed-versus-calculated mass difference. Check isoform recognition and band specificity before assigning the difference to a modification.
No glycosylation sites, signal peptide, or propeptide are listed. Those features therefore provide no basis here for attributing an unexpected band to glycosylation or precursor cleavage. Assess the listed isoforms and modified residues without assuming either produces a visible shift.
Does this guide establish induction of CCNE2?
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 CCNE2?
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 M04536 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 CCNE2 bands be quantified?
Quantitation · Specify which band or isoform is being measured, since UniProt lists Long and Short forms. Record the cell-cycle context consistently across samples because CCNE2 is annotated as a G1/S cyclin. The supplied features do not establish a particular expression change across conditions.
What should be considered for an unexpected lower band?
Interpretation · Consider the Short isoform, which lacks canonical residues 167–211. The supplied features do not establish where it migrates or prove that any lower band is CCNE2. Check whether the antibody recognizes sequences retained in Short before assigning the band.
Boster reagents
CCNE2 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 E2 CCNE2 Rabbit Monoclonal Antibody
Cat # M04536
Real WB data
Anti-Cyclin E2 CCNE2 Rabbit Monoclonal Antibody
Cat # M04536-1
Two the supplier rabbit monoclonal anti-CCNE2 antibodies are listed, each with a Western blot image from a human cell lysate: HeLa for M04536 and Jurkat for M04536-1. The supplied evidence does not show a mouse Western blot.
Which to pick: Choose M04536 for listed Human and Mouse reactivity; its Western blot image uses HeLa lysate. Choose M04536-1 for Human reactivity if its Jurkat lysate example better matches your work. Neither supplied image demonstrates mouse Western blot performance.
Source: BosterBio CCNE2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O96020.
- Human Protein Atlas. CCNE2 tissue expression.
- PMC11733543 — target-verified WB comparison
- PMC11545013 — target-verified WB comparison
- PMC3525943 — target-verified WB comparison