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- Table of Contents
E2F2 / Transcription factor E2F2 · Western blot design guide
Design a Western Blot for E2F2
Real validated E2F2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-E2F2 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
E2F2 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 | ~47.5 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 | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated E2F2 Western Blot Protocols
The M02896 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | K562 cell lysate (catalog M02896) |
| 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 | M02896; 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 E2F2 Western Blot Band Size?
E2F2 is predicted at 47.5 kDa; no empirical band is supplied, and its listed features do not demonstrate altered migration.
| Single band near 47.5 kDa | Consistent with the predicted E2F2 mass; confirm identity with controls |
| Band near 47.5 kDa in a nuclear fraction | Consistent with E2F2's nuclear location |
| Weak band in whole-cell lysate | Nuclear E2F2 recovery or abundance may be low |
| Multiple bands | Only one isoform is listed; the additional bands need identity checks |
💡Expected E2F2 appearanceE2F2 has a predicted mass of 47.5 kDa, with no empirical band size supplied; confirm a band near that size using band-identity controls.
| UniProt predicted mass | Places the expected band near 47.5 kDa |
| Predicted molecular weight in daltons | 47,506 Da corresponds to approximately 47.5 kDa |
| Sequence-based mass estimate | Provides an expected size rather than measured gel migration |
| Predicted monomer mass | Provides the 47.5 kDa reference for evaluating a candidate band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear E2F2 may be poorly recovered | Check nuclear extraction with a nuclear marker |
| Band higher than expected | The supplied features do not establish the shift | Check band identity with E2F2 depletion |
| Band lower than expected | No listed cleavage or smaller isoform explains the band | Check band identity with E2F2 depletion |
| Broad smear instead of sharp band | No listed glycosylation site explains the smear | Check sample integrity and antibody specificity |
| Multiple bands | Only one isoform is listed, leaving band identities uncertain | Compare bands after E2F2 depletion |
| Weak or no signal | Nuclear target recovery or abundance may be low | Check nuclear extraction and use a positive lysate control |
Experiment evidence
Western blot images showing E2F2 band
M02896 · E2F2
PA1810 · E2F2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for E2F2 Western blot
🧪For positive controls for E2F2 in Western blot, you can use a sample with independently confirmed E2F2 expression; the supplied HPA evidence identifies no positive tissue or cell line.
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: Appendix offers a negative tissue, but the lack of an HPA positive candidate makes a positive control harder to select.
HPA tissue expression evidence for E2F2
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 E2F2 Western Blot Tips
Deeper troubleshooting and optimisation questions for E2F2, answered from its protein features.
Where should the E2F2 band migrate?
Band shift · The predicted mass is 47.5 kDa. No observed band position is supplied, so use 47.5 kDa as a reference rather than an expected apparent mass.
Do annotated isoforms predict additional E2F2 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Could phosphorylation affect E2F2 migration?
PTM · E2F2 has a phosphoprotein keyword, but no modified residues or site coordinates are listed. This annotation alone cannot establish a visible shift or explain a difference from 47.5 kDa.
Does this guide establish induction of E2F2?
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 E2F2?
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 M02896 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 E2F2 bands be quantified across samples?
Quantitation · Because E2F2 is nuclear, use the same sample fraction for each comparison and normalize for loading. Quantify bands within the assay’s linear signal range.
Which sample fraction is relevant for E2F2 detection?
Interpretation · E2F2 is annotated as nuclear. A nuclear fraction is therefore a relevant sample when assessing its detection; compare equivalent fractions across samples.
E2F2 forms heterodimers with DP family members and binds hypophosphorylated RB1. These interactions alone do not identify an extra band as E2F2 or predict its apparent mass.
The supplied features list one isoform, no glycosylation sites, and no signal peptide or propeptide. They do not assign identities to additional bands. Check band specificity before attributing one to E2F2 or phosphorylation.
E2F2 forms heterodimers with DP family members and binds hypophosphorylated RB1. These interactions alone do not identify an extra band as E2F2 or predict its apparent mass.
The supplied features list one isoform, no glycosylation sites, and no signal peptide or propeptide. They do not assign identities to additional bands. Check band specificity before attributing one to E2F2 or phosphorylation.
Boster reagents
E2F2 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-E2F2 Rabbit Monoclonal Antibody
Cat # M02896
Real WB data
Anti-Transcription factor E2F2 E2F2 Antibody Picoband®
Cat # PA1810
Two the supplier anti-E2F2 antibodies list human, mouse, and rat reactivity and have Western blot images. M02896 is shown with K562 lysate; PA1810 is shown with human cell and rat and mouse thymus lysates. These images document specific tested samples.
Which to pick: Choose M02896 for a K562 example or PA1810 for documented HeLa, A431, rat thymus, or mouse thymus examples. PA1810 reports a band near the expected 48 kDa under its stated conditions; both products have WB images.
Source: BosterBio E2F2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14209.
- Human Protein Atlas. E2F2 tissue expression.
- PMC5976971 — target-verified WB comparison
- PMC8056345 — target-verified WB comparison
- PMC6138916 — target-verified WB comparison