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- Table of Contents
ADGRE1 / Adhesion G protein-coupled receptor E1 · Western blot design guide
Design a Western Blot for ADGRE1
Real validated ADGRE1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ADGRE1 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
ADGRE1 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 | ~97.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +2 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Peptide-blocking control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated ADGRE1 Western Blot Protocols
The A30787 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cells (catalog A30787) |
| Gel % | 8–10% (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 | A30787; 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 ADGRE1 Western Blot Band Size?
ADGRE1 is predicted at 97.7 kDa; glycosylation, signal-peptide cleavage, and isoforms could affect migration, but no observed band or feature-driven shift is established.
| Band near 97.7 kDa | Compatible with the predicted full-length mass; identity requires controls |
| Band above 97.7 kDa | N-linked glycosylation could affect migration; the band identity and cause require testing |
| Band slightly below 97.7 kDa | Signal-peptide removal could modestly reduce mass |
| Several bands at different positions | Isoforms 1–5 could contribute, but distinct migration is unproven |
💡Expected ADGRE1 appearanceADGRE1 has a predicted full-length mass of 97.7 kDa; glycosylation, signal-peptide cleavage, and isoforms may affect migration, but no empirical band size is supplied, so verify band identity with controls.
| Predicted full-length mass | 97.7 kDa provides a sequence-based reference, not a measured band position |
| N-linked glycosylation at Asn94, Asn99, Asn127, Asn167, Asn189, Asn194, Asn232, and Asn258 | Could alter apparent size; site annotations alone do not establish the shift |
| Signal peptide at residues 1–20 | Cleavage could modestly lower mature protein mass relative to the precursor |
| Isoforms 1, 2, 3, 4, and 5 | Sequence differences could affect size, but their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ADGRE1 is a multi-pass membrane protein that may be poorly recovered | Check membrane-protein extraction and include a suitable positive lysate |
| Band higher than expected | N-linked glycosylation could affect migration | Compare matched samples before and after PNGase F treatment and verify band identity |
| Band lower than expected | Signal-peptide removal could modestly reduce mass; a large difference remains unexplained | Check antibody specificity with a positive control and confirm the detected species |
| Broad smear instead of sharp band | Variation in N-linked glycosylation is possible but unproven | Compare PNGase F-treated and untreated samples with an identity control |
| Multiple bands | Isoforms 1–5 could contribute, but distinct bands are not established | Use an ADGRE1-specific control and compare defined isoform samples if available |
Experiment evidence
Western blot images showing ADGRE1 band
A30787 · ADGRE1
A30787 · ADGRE1
A30787 · ADGRE1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ADGRE1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ADGRE1 in Western blot, you can use bone marrow lysate.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, ADGRE1 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for ADGRE1
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | Medium | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADGRE1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ADGRE1, answered from its protein features.
What should I check if unexpected ADGRE1 bands appear?
Band shift · Check the antibody epitope against the annotated isoform deletions, then consider the N-linked glycosylation sites, signal peptide at 1–20, and disulfide bonds. ADGRE1 is a multi-pass cell membrane protein. These features provide possibilities to investigate, but none identifies an unexpected band on its own.
Could ADGRE1 isoforms produce bands of different sizes?
Isoforms · Five isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 599–663; isoform 3 lacks 80–131 and has a replacement beginning at 764; isoform 4 lacks 140–316; and isoform 5 lacks 80–220. These sequence differences can affect size, but they do not establish which isoforms are present in a sample.
Compare its stated epitope with the UniProt canonical regions missing from isoforms 2–5. An epitope within a deleted region may miss that isoform. Isoform 3 also has a sequence replacement beginning at position 764. Verify the antibody’s numbering convention before comparing coordinates.
Compare its stated epitope with the UniProt canonical regions missing from isoforms 2–5. An epitope within a deleted region may miss that isoform. Isoform 3 also has a sequence replacement beginning at position 764. Verify the antibody’s numbering convention before comparing coordinates.
Which ADGRE1 glycosylation sites could affect band interpretation?
PTM · The annotated N-linked sites are at positions 94, 99, 127, 167, 189, 194, 232, 258, 312, 366, 375, and 448 in UniProt canonical numbering. Check whether an antibody targets a region containing these sites when interpreting bands; antibody or paper numbering may differ.
Does this guide establish induction of ADGRE1?
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 ADGRE1?
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 A30787 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 I quantify multiple ADGRE1 bands?
Quantitation · Define which band or bands are being measured and apply the same choice across samples. ADGRE1 has five annotated isoforms and 12 N-linked glycosylation sites, so bands may represent different molecular forms. Band position alone cannot identify an isoform or glycosylation state.
Why might ADGRE1 migrate differently from its predicted 97.7 kDa?
Interpretation · ADGRE1 has a signal peptide at residues 1–20 and 12 annotated N-linked glycosylation sites. Processing and glycosylation could affect apparent mass, but these features alone do not establish a visible shift. No observed band size is supplied for comparison.
UniProt lists 20 disulfide bonds. Compare samples prepared under the same reducing conditions when assessing migration, since disulfide status may affect how the protein runs. The annotation alone does not predict a particular band position.
UniProt lists 20 disulfide bonds. Compare samples prepared under the same reducing conditions when assessing migration, since disulfide status may affect how the protein runs. The annotation alone does not predict a particular band position.
Boster reagents
ADGRE1 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-EMR1 Antibody
Cat # A30787
A30787 is an anti-EMR1 (ADGRE1) antibody with reported human, mouse, and rat reactivity. Its supplied Western blot image uses Jurkat cells; no mouse or rat Western blot example is provided.
Which to pick: A30787 is the only listed option. It has a Jurkat cell Western blot image; for mouse or rat samples, reactivity is reported but no matching Western blot example is supplied.
Source: BosterBio ADGRE1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14246.
- Human Protein Atlas. ADGRE1 tissue expression.
- PMC9775542 — target-verified WB comparison
- PMC12363508 — target-verified WB comparison