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- Table of Contents
EZH1 / Histone-lysine N-methyltransferase EZH1 · Western blot design guide
Design a Western Blot for EZH1
Real validated EZH1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-EZH1 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
EZH1 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 | ~85.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Colon (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 | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated EZH1 Western Blot Protocols
The A02494 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse lung tissue lysate (catalog A02494) |
| 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 | A02494 · (A) 1 and (B) 2 μg/mL (catalog A02494) |
| 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 EZH1 Western Blot Band Size?
EZH1 is predicted at 85.3 kDa; five isoforms could affect migration, but distinct bands or shifts have not been demonstrated here.
| Single band near 85.3 kDa | Consistent with the predicted EZH1 size; confirm identity with controls |
| Band away from 85.3 kDa | An EZH1 isoform is possible, but its migration is unknown |
| Doublet | Could reflect isoforms 1–5 if they migrate separately; confirm both bands |
| Several distinct bands | Could reflect separately migrating isoforms; band identities require validation |
💡Expected EZH1 appearanceEZH1 has a predicted mass of 85.3 kDa, but no empirical band size or isoform migration is supplied; use band-identity controls to assess bands near that size or at other positions.
| Predicted EZH1 mass | The 747-residue sequence has a predicted mass of 85.3 kDa |
| Isoform 1 | Its individual apparent size is not supplied |
| Isoforms 2 and 3 | Alternative splicing could change migration, but their individual sizes are not supplied |
| Isoforms 4 and 5 | Alternative splicing could change migration, but their individual sizes are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear EZH1 may be poorly represented in the prepared lysate | Check nuclear extraction and a nuclear protein control |
| Band higher than expected | An isoform is possible, but its migration is unknown | Confirm identity with an independent EZH1 antibody or EZH1 depletion |
| Band lower than expected | An isoform is possible, but its migration is unknown | Confirm identity with EZH1 depletion and check sample integrity |
| Multiple bands | Isoforms 1–5 could contribute if they migrate separately | Validate each band with EZH1 depletion or an independent antibody |
| Weak or no signal | The lysate may contain little nuclear EZH1 | Check nuclear enrichment and antibody performance with a positive control |
Experiment evidence
Western blot images showing EZH1 band
A02494 · EZH1
A02494-1 · EZH1
A02494-1 · EZH1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EZH1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for EZH1 in Western blot, you can use colon tissue, which shows high expression in HPA.
Positive control: Colon (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: A negative tissue is available, but EZH1’s nuclear location may make nuclear-enriched lysates useful for detection.
HPA tissue expression evidence for EZH1
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 |
|---|---|---|---|---|
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | glands of Brunner | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular 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 → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced EZH1 Western Blot Tips
Deeper troubleshooting and optimisation questions for EZH1, answered from its protein features.
How should EZH1 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 EZH1 isoforms produce different bands?
Isoforms · Five isoforms are listed. Relative to the canonical sequence, isoform 3 lacks residues 83–122, isoform 4 lacks residues 1–70, and isoform 5 replaces residues 1–162 with a shorter sequence. Isoform 2 changes the first residue to MEDYSKM. These differences could affect apparent size, but no isoform band positions are supplied.
Check its epitope against the sequence changes before interpreting missing bands. Canonical residues 1–70 are absent from isoform 4, residues 1–162 are replaced in isoform 5, and residues 83–122 are absent from isoform 3. These are UniProt canonical coordinates; antibody numbering may differ.
It is possible, given the five listed isoforms and their sequence differences. Compare the band with the antibody’s epitope and the specific sequence changes. No observed band positions are supplied, so an unexpected band cannot be assigned to an isoform from these features alone.
Check its epitope against the sequence changes before interpreting missing bands. Canonical residues 1–70 are absent from isoform 4, residues 1–162 are replaced in isoform 5, and residues 83–122 are absent from isoform 3. These are UniProt canonical coordinates; antibody numbering may differ.
It is possible, given the five listed isoforms and their sequence differences. Compare the band with the antibody’s epitope and the specific sequence changes. No observed band positions are supplied, so an unexpected band cannot be assigned to an isoform from these features alone.
Do the listed modifications explain an EZH1 band shift?
PTM · The keywords include Ubl conjugation and isopeptide bond, but the supplied features list no modified residue coordinates. They do not identify a modified band or demonstrate a visible shift.
Does this guide establish induction of EZH1?
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 EZH1?
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 A02494 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.
Does H3K27me3 signal quantify EZH1 abundance?
Quantitation · EZH1 is nuclear and colocalizes with histone H3 trimethylated at Lys-27. Colocalization alone does not establish that H3K27me3 signal is proportional to EZH1 protein abundance. Quantify the EZH1 band directly when measuring EZH1.
Should EZH1 run at exactly 85.3 kDa?
Interpretation · 85.3 kDa is the predicted mass of canonical EZH1. No observed band position is supplied, so use it as a reference rather than an exact expected position. The listed features do not establish a visible shift.
Boster reagents
EZH1 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-EZH1 Antibody
Cat # A02494
Real WB data
Anti-EZH1 Antibody Picoband®
Cat # A02494-1
Two anti-EZH1 antibodies list human, mouse, and rat reactivity and include WB images. A02494 is shown with mouse lung lysate; A02494-1 is shown with human cell lysates. The latter caption reports bands near 100 and 60 kDa, versus an expected 68 kDa.
Which to pick: For a mouse lung lysate example, see A02494 at 1–2 μg/mL. For human cell lysate examples, see A02494-1 at 0.5 μg/mL. Both have WB images; neither supplied caption shows a rat sample.
Source: BosterBio EZH1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92800.
- Human Protein Atlas. EZH1 tissue expression.
- PMC11699051 — target-verified WB comparison
- PMC6743615 — target-verified WB comparison
- PMC5346642 — target-verified WB comparison