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- Table of Contents
ESD / S-formylglutathione hydrolase · Western blot design guide
Design a Western Blot for ESD
Real validated ESD Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ESD 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
ESD 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 | ~31.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ESD Western Blot Protocols
The A01471 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysates from Jurkat, K562 cell line, mouse stomach and rat stomach tissue (from left to right), (catalog A01471) |
| 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 | A01471; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:5000 (catalog A01471) |
| 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 ESD Western Blot Band Size?
ESD has a predicted 31.5 kDa monomer; its homodimer could affect migration, but no empirical band or visible shift is established.
| Single band near 31.5 kDa | consistent with the predicted ESD monomer; confirm band identity |
| Band near 63 kDa | could reflect a homodimer that remained associated during preparation |
| Band near 31.5 kDa in cytoplasmic fraction | consistent with ESD's cytoplasmic location |
| Band near 31.5 kDa in vesicle fraction | consistent with ESD's cytoplasmic vesicle location |
💡Expected ESD appearanceThe predicted ESD monomer is 31.5 kDa; no empirical band size is supplied, so confirm a candidate band with ordinary identity controls.
| Predicted monomer mass | places the expected monomer near 31.5 kDa |
| Homodimer | could appear near twice the monomer mass if it remains associated |
| N-acetylalanine at residue 2 | has no demonstrated visible effect on migration |
| N6-succinyllysine at residue 4 | has no demonstrated visible effect on migration |
| N6-acetyllysine at residue 200 | has no demonstrated visible effect on migration |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | insufficient recovery of cytoplasmic or vesicular ESD | check extraction and compare whole-cell and vesicle-enriched samples |
| Band higher than expected | the ESD homodimer may remain associated | compare denaturing conditions and check band identity |
| Band lower than expected | the listed features do not establish a smaller ESD product | verify identity with a second antibody or ESD depletion |
| Multiple bands | a persistent homodimer or unrelated antibody binding may contribute | compare denaturing conditions and verify bands by ESD depletion |
| Weak or no signal | ESD recovery may vary between cytoplasmic and vesicle fractions | check fraction recovery and sample loading |
Experiment evidence
Western blot images showing ESD band
A01471 · ESD
A01471-1 · ESD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ESD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ESD in Western blot, you can use adrenal gland tissue, which HPA scores High.
Positive control: Adrenal gland (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: Cytoplasmic ESD is suitable for lysate controls, but verify the HPA not-detected adipose tissue in your assay.
HPA tissue expression evidence for ESD
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | 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 → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ESD Western Blot Tips
Deeper troubleshooting and optimisation questions for ESD, answered from its protein features.
Where should the main ESD band appear?
Band shift · The predicted mass is 31.5 kDa for the 282-residue protein. No empirical band position is supplied, so compare the observed band with this estimate without assuming an exact match.
Could ESD isoforms produce multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not support assigning multiple bands to annotated ESD isoforms.
Which ESD modifications matter when interpreting band mobility?
PTM · UniProt lists N-acetylalanine at position 2, N6-succinyllysine at position 4, and N6-acetyllysine at position 200. These are UniProt coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible shift or explain a mass discrepancy.
No glycosylation sites are listed for ESD, so the supplied features do not support assigning a higher band to glycosylation.
No glycosylation sites are listed for ESD, so the supplied features do not support assigning a higher band to glycosylation.
Does this guide establish induction of ESD?
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 ESD?
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 A01471 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 ESD be quantified across cell fractions?
Quantitation · ESD is listed in the cytoplasm and cytoplasmic vesicles. If comparing fractions, quantify each band against an appropriate loading reference for that fraction and keep fraction preparation consistent. A signal in either location is compatible with the supplied localization.
Could processing explain a smaller ESD band?
Interpretation · No signal peptide or propeptide is listed, so the supplied features do not support a predicted cleavage product. Investigate a smaller band before assigning it to ESD.
ESD is annotated as a homodimer. Two 31.5 kDa subunits suggest about 63 kDa before considering gel behavior, but the annotation does not establish that a dimer persists during Western blot preparation. Verify a higher band before assigning it to ESD.
ESD is annotated as a homodimer. Two 31.5 kDa subunits suggest about 63 kDa before considering gel behavior, but the annotation does not establish that a dimer persists during Western blot preparation. Verify a higher band before assigning it to ESD.
Boster reagents
ESD 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-ESD Antibody (Center)
Cat # A01471
Real WB data
Anti-ESD Antibody Picoband®
Cat # A01471-1
A01471 and A01471-1 list human, mouse, and rat reactivity and each includes a Western blot image. A01471-1 reports an ESD band near 31 kDa. The supplied evidence consists of product blot captions; no publication evidence was provided.
Which to pick: Choose A01471 for the shown Jurkat, K562, or mouse and rat stomach lysates. Choose A01471-1 for the shown 293T, HEL, K562, HepG2, or mouse and rat kidney and liver lysates. Both have WB images; these specimen examples do not guarantee other contexts.
Source: BosterBio ESD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P10768.
- Human Protein Atlas. ESD tissue expression.
- PMC3796443 — target-verified WB comparison
- PMC8903700 — target-verified WB comparison