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- Table of Contents
ESRRA / Steroid hormone receptor ERR1 · Western blot design guide
Design a Western Blot for ESRRA
Real validated ESRRA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ESRRA 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
ESRRA 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 | ~45.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bronchus (IHC candidate; verify WB) +4 more | |
| Negative control | Oral mucosa (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated ESRRA Western Blot Protocols
The M03339 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 M03339) |
| 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 | M03339; 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 ESRRA Western Blot Band Size?
ESRRA has a predicted mass of 45.5 kDa; isoforms and listed modifications could affect migration, but no empirical band size or visible shift is established.
| Band near 45.5 kDa | Consistent with the predicted ESRRA mass; confirm identity with controls. |
| Closely spaced doublet | Could reflect modified ESRRA, though its listed sites do not establish a visible shift. |
| Several bands at different positions | Could include isoforms 1 and 2; their migration differences are unknown. |
| Weak band in a cytoplasmic fraction | Consistent with predominantly nuclear localization; cytoplasmic localization is reported with MAPK15. |
💡Expected ESRRA appearanceUniProt predicts 45.5 kDa for ESRRA; no empirical band size is supplied, and any additional bands require identity controls because isoform migration and modification effects are unquantified.
| UniProt predicted mass | Places the reference band near 45.5 kDa. |
| Isoforms 1 and 2 | May differ in apparent size, but their masses and migration are not supplied. |
| Phosphoserines at residues 19 and 22 | Could alter migration; no visible shift is established. |
| Acetyllysines at residues 129, 138, 160 and 162 | Could alter migration; no visible shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear ESRRA may be underrepresented in the preparation. | Check nuclear extraction and include a known positive sample. |
| Band higher than expected | A modification-related shift is possible but unproven. | Compare with a positive sample and verify specificity by ESRRA depletion. |
| Band lower than expected | An isoform or protein breakdown is possible; neither is established by band position alone. | Compare fresh preparations and verify the band by ESRRA depletion. |
| Multiple bands | Isoforms 1 and 2 or modified forms may contribute. | Use ESRRA depletion to identify specific bands. |
| Weak or no signal | Low recovery of predominantly nuclear ESRRA is possible. | Check nuclear fraction recovery and antibody performance with a positive sample. |
| Fragments below expected size | Protein breakdown during preparation is possible. | Prepare fresh lysate with protease inhibitors and compare band patterns. |
Experiment evidence
Western blot images showing ESRRA band
M03339 · ESRRA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ESRRA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ESRRA in Western blot, you can use bronchus tissue, which HPA rates as high expression.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Oral mucosa (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: ESRRA is mainly nuclear, so nuclear-enriched lysates may improve detection.
HPA tissue expression evidence for ESRRA
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 |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ESRRA Western Blot Tips
Deeper troubleshooting and optimisation questions for ESRRA, answered from its protein features.
How should ESRRA 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 ESRRA isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2, with position 191 missing in isoform 2. An antibody recognizing both could detect both, but this sequence difference alone does not establish that two bands will resolve. Check which isoforms the antibody recognizes.
Decide whether the measurement represents isoform 1, isoform 2, or both. Isoform 2 lacks UniProt position 191, so antibody coverage and band assignment matter. Quantify the same assigned band or set of bands consistently across samples.
Decide whether the measurement represents isoform 1, isoform 2, or both. Isoform 2 lacks UniProt position 191, so antibody coverage and band assignment matter. Quantify the same assigned band or set of bands consistently across samples.
Which ESRRA phosphorylation sites matter for band interpretation?
PTM · UniProt lists phosphoserine at positions 19 and 22. If comparing phospho-specific and total ESRRA signals, verify that the antibody targets the intended site. These are UniProt coordinates, which may differ from antibody or paper numbering. Phosphorylation at these sites does not by itself establish a visible shift.
PCAF/KAT2B acetylates lysines 129, 138, 160, and 162 according to the supplied features. If an antibody epitope overlaps these residues, compare its recognition of modified and unmodified ESRRA when interpreting signal changes. The positions use UniProt numbering; check the numbering convention in antibody documentation.
The supplied features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for assigning a higher band to glycosylation or precursor processing. Compare candidate bands with the 45.5 kDa prediction and verify ESRRA identity experimentally.
PCAF/KAT2B acetylates lysines 129, 138, 160, and 162 according to the supplied features. If an antibody epitope overlaps these residues, compare its recognition of modified and unmodified ESRRA when interpreting signal changes. The positions use UniProt numbering; check the numbering convention in antibody documentation.
The supplied features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for assigning a higher band to glycosylation or precursor processing. Compare candidate bands with the 45.5 kDa prediction and verify ESRRA identity experimentally.
Does this guide establish induction of ESRRA?
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 ESRRA?
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 M03339 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 ESRRA 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.
Where should ESRRA appear relative to its predicted mass?
Interpretation · The supplied predicted mass is 45.5 kDa, but no observed Western blot band is provided. Use 45.5 kDa as a reference, not a guaranteed migration position; the listed features alone cannot establish a band shift.
ESRRA is listed in the nucleus and in the cytoplasm, with cytoplasmic co-localization only in the presence of MAPK15. When quantifying fractions, compare like fractions across conditions and account for MAPK15 status. A change in one fraction need not indicate a change in total ESRRA.
ESRRA binds DNA as a monomer or homodimer, but that functional feature alone does not identify a Western blot band as a dimer. Check the band's reproducibility and antibody specificity before assigning it to an ESRRA complex.
ESRRA is listed in the nucleus and in the cytoplasm, with cytoplasmic co-localization only in the presence of MAPK15. When quantifying fractions, compare like fractions across conditions and account for MAPK15 status. A change in one fraction need not indicate a change in total ESRRA.
ESRRA binds DNA as a monomer or homodimer, but that functional feature alone does not identify a Western blot band as a dimer. Check the band's reproducibility and antibody specificity before assigning it to an ESRRA complex.
Boster reagents
ESRRA 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-ERR alpha Rabbit Monoclonal Antibody
Cat # M03339
M03339 is an anti-ESRRA rabbit monoclonal antibody listed for Western blotting, with reported Human, Mouse, and Rat reactivity. Its WB image shows ERR alpha expression in HeLa cell lysate; the supplied evidence does not document other tested specimens.
Which to pick: M03339 is the only listed option. Its WB image uses HeLa cell lysate, so it is the most directly documented choice for that sample; check suitability for other samples despite the listed Human, Mouse, and Rat reactivity.
Source: BosterBio ESRRA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P11474.
- Human Protein Atlas. ESRRA tissue expression.
- PMC12003256 — target-verified WB comparison
- PMC11069533 — target-verified WB comparison