This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ERH / Enhancer of rudimentary homolog · Western blot design guide
Design a Western Blot for ERH
Source-linked ERH Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ERH 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
ERH 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 | ~12.3 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Smooth muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ERH Western Blot Protocol Options
The A05129 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A05129) |
| Gel % | 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 | 0.2 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05129) |
| Primary antibody | A05129 · 1:3000 (catalog A05129) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05129) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05129) |
Section 2
What Is the Expected ERH Western Blot Band Size?
ERH predicts a 12.3 kDa monomer; homodimer retention or phosphorylation could affect the pattern, but no empirical migration is supplied.
| Band near 12.3 kDa | Consistent with the predicted ERH monomer; confirm its identity with antibody controls. |
| Band near 25 kDa | Could reflect a retained ERH homodimer, which UniProt annotates by similarity. |
| Close doublet near 12.3 kDa | Could reflect different phosphorylation states; a visible shift is not established. |
| Weak band in a cytoplasmic fraction | Consistent with ERH's nuclear location; check a nuclear fraction. |
💡Expected ERH appearanceERH has a predicted monomer mass of 12.3 kDa, but no empirical band size is supplied; confirm any candidate band with nuclear enrichment and antibody specificity controls.
| Predicted ERH mass | Places the monomer near 12.3 kDa before considering migration effects. |
| Homodimer by similarity | Could yield a band near twice the monomer size if the dimer survives sample preparation. |
| N-acetylserine at residue 2 | Is a documented modification, with no demonstrated visible size shift. |
| Phosphothreonine at residue 11 | Could affect migration, but no resolvable band shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ERH may be poorly recovered during extraction. | Check the nuclear fraction and an extraction control. |
| Band higher than expected | A homodimer may persist during sample preparation. | Compare fully denatured samples and verify band identity. |
| Band lower than expected | A lower band may be unrelated to the predicted ERH monomer. | Check antibody specificity and compare with a nuclear fraction. |
| Multiple bands | Monomer and retained homodimer are possible; phosphorylation may also affect migration. | Compare sample preparation conditions and verify each band with specificity controls. |
| Weak or no signal | The extraction may recover little nuclear ERH. | Assess nuclear extraction and include a positive lysate control. |
Experiment evidence
Western blot images showing ERH band
A05129 · ERH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ERH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ERH in Western blot, you can use adipose tissue, which HPA rates as highly expressed.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Smooth muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Tissue controls are feasible, though ERH’s nuclear location may favor nuclear-enriched lysate.
HPA tissue expression evidence for ERH
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in white pulp | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ERH Western Blot Tips
Deeper troubleshooting and optimisation questions for ERH, answered from its protein features.
How should ERH 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 ERH isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands; verify their identity before assigning them to ERH.
Which ERH modifications matter when interpreting a band?
PTM · UniProt lists N-acetylserine at residue 2 and phosphothreonine at residue 11. These are UniProt sequence coordinates; antibody or paper numbering may differ. Neither annotation establishes a detectable band shift.
Does this guide establish induction of ERH?
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 ERH Western blot?
Transfer · ERH is small, at 104 residues and a predicted 12.3 kDa. Choose and verify transfer conditions that retain a protein near this size. The supplied features do not identify a specific membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05129 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 ERH 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.
Should ERH migrate at its predicted 12.3 kDa?
Interpretation · ERH is 104 residues with a predicted mass of 12.3 kDa. No observed band position is supplied, so use 12.3 kDa as a reference rather than an exact migration requirement. Its annotated modifications alone do not establish a visible shift.
ERH is annotated as a homodimer by similarity and as a methylosome component. Those annotations do not establish that a dimer or complex survives Western blot preparation or explain a higher band. Confirm band identity before making that assignment.
ERH is annotated in the nucleus. Include an appropriate nuclear sample when assessing its presence, and keep sample fractions consistent across comparisons.
Compare ERH in the same type of sample fraction and at the same assigned band, using consistent loading and detection. Its nuclear localization makes fraction differences relevant; the supplied features provide no basis to infer changes in abundance from a band shift.
ERH is annotated as a homodimer by similarity and as a methylosome component. Those annotations do not establish that a dimer or complex survives Western blot preparation or explain a higher band. Confirm band identity before making that assignment.
ERH is annotated in the nucleus. Include an appropriate nuclear sample when assessing its presence, and keep sample fractions consistent across comparisons.
Compare ERH in the same type of sample fraction and at the same assigned band, using consistent loading and detection. Its nuclear localization makes fraction differences relevant; the supplied features provide no basis to infer changes in abundance from a band shift.
Boster reagents
ERH 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-ERH Antibody
Cat # A05129
A05129 is listed as human-reactive and has a Western blot image using cell-line extracts, 25 µg protein per lane, and a 1:3000 antibody dilution. The caption does not name the cell lines.
Which to pick: A05129 is the only listed ERH antibody. Choose it for human ERH Western blots; its product record includes a Western blot image and reported assay conditions.
Source: BosterBio ERH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P84090.
- Human Protein Atlas. ERH tissue expression.
- PMC8797358 — target-verified WB comparison
- PMC4794255 — target-verified WB comparison
- PMC13366317 — target-verified WB comparison