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- Table of Contents
ESRRB / Steroid hormone receptor ERR2 · Western blot design guide
Design a Western Blot for ESRRB
Real validated ESRRB Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ESRRB 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
ESRRB 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 | ~48.1 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 | Acetylated | |
| Caveat | Blocking peptide control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated ESRRB Western Blot Protocols
The A03473 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human heart tissue lysate (catalog A03473) |
| 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 | Western blot analysis of ESRRB in human heart tissue lysate with ESRRB antibody at 1 μg/mL in (A) the absence and (B) the presence of blocking peptide (catalog A03473) |
| Primary antibody | A03473 · 1 μg/mL (catalog A03473) |
| 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 ESRRB Western Blot Band Size?
ESRRB is predicted at 48.1 kDa; three isoforms may affect migration, but no distinct isoform bands or empirical band size are established.
| Band near 48.1 kDa | Consistent with the predicted ESRRB mass; identity needs confirmation |
| Several bands at different sizes | Isoforms 1, 2, and 3 are possible contributors; their separation is unestablished |
| Band in a nuclear fraction | Consistent with ESRRB nuclear localization |
| Band in a cytoplasmic fraction | Consistent with ESRRB cytoplasmic localization |
💡Expected ESRRB appearanceESRRB has a predicted mass of 48.1 kDa, but no empirical band size is supplied; use antibody blocking peptide or another band-identity control to assess any detected band.
| Predicted ESRRB mass | Sets a 48.1 kDa sequence-based reference, not a measured band position |
| Splice isoform 1 | May differ in apparent size from other isoforms; its mass is unspecified |
| Splice isoform 2 | May differ in apparent size from other isoforms; its mass is unspecified |
| Splice isoform 3 | May differ in apparent size from other isoforms; its mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear localization may limit signal in whole-cell lysate | Check a nuclear fraction and a suitable positive control |
| Band higher than expected | An isoform may migrate differently, but no isoform masses are supplied | Test band identity with blocking peptide or an independent antibody |
| Band lower than expected | An isoform may migrate differently, but no isoform masses are supplied | Test band identity with blocking peptide or an independent antibody |
| Multiple bands | Three splice isoforms exist, although distinct bands are unconfirmed | Check which bands are lost with blocking peptide |
| Weak or no signal | ESRRB may be diluted across whole-cell lysate because it localizes to the nucleus | Enrich the nuclear fraction and include a positive control |
Experiment evidence
Western blot images showing ESRRB band
A03473 · ESRRB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ESRRB Western blot
🧪For positive controls for ESRRB in Western blot, you can use an independently validated ESRRB-positive cell lysate because no HPA positive sample is supplied.
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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA provides no tissue or cell candidates, so a validated positive sample and an ESRRB knockout control would need to be sourced separately.
HPA tissue expression evidence for ESRRB
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 ESRRB Western Blot Tips
Deeper troubleshooting and optimisation questions for ESRRB, answered from its protein features.
How should ESRRB 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 ESRRB isoforms produce different bands?
Isoforms · Three isoforms are listed. In isoforms 1 and 2, residue 433 is replaced by a different C-terminal sequence in each case. These changes could affect protein size or antibody recognition, but the features do not establish where their bands migrate.
Check whether the antibody epitope includes residue 433 or the C-terminal sequence that differs in isoforms 1 and 2. An antibody against a shared region is better suited to detecting all three listed isoforms.
Check whether the antibody epitope includes residue 433 or the C-terminal sequence that differs in isoforms 1 and 2. An antibody against a shared region is better suited to detecting all three listed isoforms.
Do annotated modifications explain a shifted ESRRB band?
PTM · Acetylation appears among the keywords, but no modified residues or glycosylation sites are listed. The supplied features therefore do not establish a modification responsible for a visible shift or an observed-versus-predicted mass difference.
Does this guide establish induction of ESRRB?
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 ESRRB?
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 A03473 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 can I make ESRRB band quantitation comparable?
Quantitation · Compare bands from the same cellular fraction across samples, since ESRRB has nuclear, cytoplasmic and chromosome annotations. If multiple bands appear, quantify a consistently identified band; the supplied features do not establish which band represents a particular isoform.
What molecular weight should I expect for ESRRB?
Interpretation · The supplied sequence is 433 amino acids with a predicted mass of 48.1 kDa. No observed band position is available, so use 48.1 kDa as a reference rather than an established apparent mass.
ESRRB is annotated in the nucleus, with cytoplasm and chromosome also listed. A nuclear fraction is a sensible starting point; keep the fraction consistent when comparing samples.
Consider the three listed isoforms and whether the antibody recognizes their differing C termini. Compare any extra bands with the 48.1 kDa sequence-based reference, but do not assign a band to an isoform or modification from these features alone.
ESRRB is annotated in the nucleus, with cytoplasm and chromosome also listed. A nuclear fraction is a sensible starting point; keep the fraction consistent when comparing samples.
Consider the three listed isoforms and whether the antibody recognizes their differing C termini. Compare any extra bands with the 48.1 kDa sequence-based reference, but do not assign a band to an isoform or modification from these features alone.
Boster reagents
ESRRB 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-Steroid hormone receptor ERR2 ESRRB Antibody
Cat # A03473
The catalog reports A03473, an anti-ESRRB antibody with reported human reactivity. Its WB image shows human heart tissue lysate tested at 1 μg/mL, with and without blocking peptide. The supplied evidence covers this sample and condition only.
Which to pick: A03473 is the only listed ESRRB antibody. It has a WB image using human heart tissue lysate; choose it when that reported context fits your experiment, and verify performance in other samples independently.
Source: BosterBio ESRRB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95718.
- Human Protein Atlas. ESRRB tissue expression.
- PMC3968251 — target-verified WB comparison
- PMC4525203 — target-verified WB comparison
- PMC3953728 — target-verified WB comparison