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- Table of Contents
ERAP1 / Endoplasmic reticulum aminopeptidase 1 · Western blot design guide
Design a Western Blot for ERAP1
Source-linked ERAP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ERAP1 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
ERAP1 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 | ~107.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked ERAP1 Western Blot Protocol Options
The M02021-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | K562 cell lysate (catalog M02021-1) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02021-1; 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 ERAP1 Western Blot Band Size?
ERAP1 is predicted at 107.2 kDa; glycosylation and isoforms could affect migration, but no apparent band size or distinct pattern is demonstrated.
| Band near 107.2 kDa | Consistent with the predicted ERAP1 mass; confirm identity with antibody controls |
| Band above 107.2 kDa | N-linked glycosylation could affect migration, but the shift is unverified |
| Broad band or smear | Variable N-linked glycosylation is possible, but its effect on this blot is unverified |
| Several bands near the expected region | Isoforms 1 and 2 could contribute, but distinct migration is unverified |
💡Expected ERAP1 appearanceERAP1 has a predicted mass of 107.2 kDa; no empirical band size is supplied, so use band-identity controls to assess bands affected by possible glycosylation or isoforms.
| UniProt predicted mass | 107.2 kDa is the reference mass, not a validated apparent band size |
| N-linked glycosylation at Asn70 | May affect apparent size; the contribution is unknown |
| N-linked glycosylation at Asn154 | May affect apparent size; the contribution is unknown |
| N-linked glycosylation at Asn414 | May affect apparent size; the contribution is unknown |
| N-linked glycosylation at Asn760 | May affect apparent size; the contribution is unknown |
| N-linked glycosylation at Asn901 | May affect apparent size; the contribution is unknown |
| Isoforms 1 and 2 | Their apparent sizes may differ; no relative order or resolved pattern is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ERAP1 may be poorly extracted | Check membrane extraction and compare an enriched membrane fraction |
| Band higher than expected | N-linked glycosylation may alter migration | Compare untreated and deglycosylated samples with an antibody specificity control |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Compare untreated and deglycosylated samples and check antibody specificity |
| Multiple bands | Isoforms 1 and 2 may contribute, but their migration is unknown | Check antibody specificity and compare samples with differing isoform expression |
| Weak or no signal | Poor recovery of membrane-associated ERAP1 is possible | Check extraction, loading, and an ERAP1-positive control |
| Band lower than expected | Its identity is unclear from the supplied features | Check antibody specificity and compare with an ERAP1-positive control |
Experiment evidence
Western blot images showing ERAP1 band
M02021-1 · ERAP1
A02021-1 · ERAP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ERAP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ERAP1 in Western blot, you can use adipose tissue, which has high HPA expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside.
⚠️Feasibility: No HPA tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for ERAP1
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 → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Salivary gland | glandular cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ERAP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ERAP1, answered from its protein features.
How should ERAP1 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 ERAP1 isoforms produce separate bands?
Isoforms · Two isoforms are listed. In isoform 2, residues 940–941 change from RM to HDPEADATG, adding seven residues near the C terminus. Check whether the antibody recognizes both isoforms before assigning bands. The sequence difference alone does not establish that they resolve separately.
Which glycosylation sites matter when assessing ERAP1 bands?
PTM · UniProt lists N-linked glycosylation at Asn 70, 154, 414, 760, and 901. These are UniProt sequence coordinates; paper or antibody numbering may differ. Compare paired untreated and deglycosylated samples to test whether glycans affect the band pattern.
Does this guide establish induction of ERAP1?
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 ERAP1 Western blot?
Transfer · ERAP1 is a predicted 107.2-kDa, single-pass type II ER membrane protein. Verify transfer of proteins around this size with a membrane stain and check the gel for retained ERAP1 signal. Optimize transfer conditions empirically; the supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02021-1 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 ERAP1 bands be quantified?
Quantitation · Use comparable sample fractions and loading across lanes, and quantify bands within a linear detection range. If multiple bands appear, define which band or bands are included consistently. The two isoforms and five glycosylation sites make band assignment important before combining signals.
How should a band above ERAP1’s predicted 107.2 kDa be interpreted?
Interpretation · ERAP1 has five annotated N-linked glycosylation sites, so compare untreated and deglycosylated samples when investigating migration. No observed band mass is supplied. Glycosylation annotations alone do not establish a visible shift or explain any measured difference.
ERAP1 is listed as a monomer that may also form a heterodimer with ERAP2. Compare reducing, denaturing conditions and confirm antibody specificity before assigning a high-mass band to a complex. The interaction annotation does not establish that a complex survives Western-blot preparation.
UniProt lists two disulfide bonds. Compare reducing and nonreducing preparations if migration differs between experiments, while keeping loading and transfer conditions comparable. The bond annotation does not predict a particular band shift.
ERAP1 is listed as a monomer that may also form a heterodimer with ERAP2. Compare reducing, denaturing conditions and confirm antibody specificity before assigning a high-mass band to a complex. The interaction annotation does not establish that a complex survives Western-blot preparation.
UniProt lists two disulfide bonds. Compare reducing and nonreducing preparations if migration differs between experiments, while keeping loading and transfer conditions comparable. The bond annotation does not predict a particular band shift.
Boster reagents
ERAP1 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-ERAP1 Rabbit Monoclonal Antibody
Cat # M02021-1
Real WB data
Anti-ERAP1 Antibody
Cat # A02021-1
Two the supplier anti-ERAP1 antibodies are listed for Western blotting, each with a product image from a named cell lysate. No publication evidence or broader validation is supplied.
Which to pick: Choose M02021-1 if rat reactivity is needed; its WB image uses K562 lysate. A02021-1 lists human and mouse reactivity and shows SK-N-SH lysate at 1 and 2 μg/ml. Both have WB images, so match the listed reactivity and sample context to your experiment.
Source: BosterBio ERAP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NZ08.
- Human Protein Atlas. ERAP1 tissue expression.
- PMC11494811 — target-verified WB comparison
- PMC9368314 — target-verified WB comparison
- PMC3759077 — target-verified WB comparison