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EPHA7 / Ephrin type-A receptor 7 · Western blot design guide
Design a Western Blot for EPHA7
Source-linked EPHA7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-EPHA7 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
EPHA7 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 | ~112.1 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked EPHA7 Western Blot Protocol Options
The A03943 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | 3T3-L1 (40ug), H9C2 (40ug), A549 (40ug), HEK293T (40ug) (catalog A03943) |
| 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 | A03943; 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 EPHA7 Western Blot Band Size?
EPHA7 has a predicted precursor mass of 112.1 kDa; cleavage, glycosylation, and isoforms may affect bands, but no migration position is demonstrated.
| Band near 112.1 kDa | Consistent with the predicted precursor mass; confirm identity with band controls |
| Band above 112.1 kDa | N-linked glycosylation at Asn343 and Asn410 could affect migration |
| Band below 112.1 kDa | Signal-peptide removal could reduce the mass relative to the precursor |
| Several bands at different positions | Isoforms 1–5 could differ in size, but distinct bands are not established |
💡Expected EPHA7 appearanceEPHA7 has a predicted precursor mass of 112.1 kDa; signal-peptide cleavage and N-linked glycosylation may affect migration, but no empirical band position is supplied, so confirm identity with appropriate controls.
| Predicted precursor mass | Provides a 112.1 kDa sequence-based reference, not a measured band position |
| N-linked glycosylation at Asn343 | May increase apparent size; the extent is not supplied |
| N-linked glycosylation at Asn410 | May increase apparent size; the extent is not supplied |
| Signal peptide at residues 1–27 | Its removal makes the mature chain smaller than the precursor |
| Isoforms 1, 2, 3, 4, and 5 | May differ in size; their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane EPHA7 may be poorly recovered during extraction | Check membrane-protein recovery and use a positive-control lysate |
| Band higher than expected | N-linked glycosylation at Asn343 and Asn410 may affect migration | Compare with a deglycosylated sample and confirm band identity |
| Band lower than expected | The signal peptide may have been removed | Check the antibody epitope and confirm identity with an independent antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but not established | Compare untreated and deglycosylated samples |
| Multiple bands | Isoforms 1–5 may contribute, but distinct migration is unverified | Check isoform expression and compare with an independent antibody |
| Weak or no signal | Recovery of this membrane receptor may be low | Check membrane extraction and include a positive-control lysate |
Experiment evidence
Western blot images showing EPHA7 band
A03943 · EPHA7
M03943 · EPHA7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EPHA7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for EPHA7 in Western blot, you can use adipose tissue, which HPA scores High.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because EPHA7 is a membrane protein, membrane-enriched lysate may improve detection.
HPA tissue expression evidence for EPHA7
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 | endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Testis | cells in seminiferous ducts | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
Section 3
Advanced EPHA7 Western Blot Tips
Deeper troubleshooting and optimisation questions for EPHA7, answered from its protein features.
How should EPHA7 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.
How could EPHA7 isoforms affect band interpretation?
Isoforms · Five isoforms are listed. Relative to the canonical 998-residue sequence, isoform 3 lacks residues 280–998 and isoform 5 lacks 451–998; isoform 2 lacks 540–544, and isoform 4 changes 600–604. Check whether the antibody epitope is present in each isoform before assigning bands. Coordinates here follow the supplied UniProt canonical sequence.
Which EPHA7 glycosylation sites matter when assessing bands?
PTM · UniProt lists N-linked sites at Asn343 and Asn410 in the canonical sequence. Glycosylation could affect migration, but site annotation alone cannot establish the cause or size of a band shift. Isoform 3 lacks both listed sites; isoform 5 retains them.
The supplied features describe ligand-bound EPHA7 as a receptor dimer within an ephrin–receptor heterotetramer and list autocatalytic phosphorylation at Tyr608, Tyr614, Tyr791 and Tyr940. Compare stimulated and baseline samples when assessing phosphorylation; confirm the response experimentally. These are UniProt canonical coordinates, which may differ from antibody or paper numbering.
The supplied features describe ligand-bound EPHA7 as a receptor dimer within an ephrin–receptor heterotetramer and list autocatalytic phosphorylation at Tyr608, Tyr614, Tyr791 and Tyr940. Compare stimulated and baseline samples when assessing phosphorylation; confirm the response experimentally. These are UniProt canonical coordinates, which may differ from antibody or paper numbering.
Does this guide establish induction of EPHA7?
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 EPHA7 Western blot?
Transfer · EPHA7 is a 998-residue, single-pass membrane protein with a predicted mass of 112.1 kDa. The supplied features do not specify a transfer method. Check transfer of proteins near this size with a total-protein stain and adjust the method if transfer is incomplete.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03943 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 EPHA7 bands be quantified across samples?
Quantitation · Quantify a consistently identified band within the assay’s linear range and normalize it across samples. Because EPHA7 has five listed isoforms, confirm that the chosen antibody recognizes the same sequence in the forms being compared. Keep a shorter isoform band separate from a full-length band.
Why might EPHA7 migrate differently from its predicted 112.1 kDa mass?
Interpretation · EPHA7 has a signal peptide at residues 1–27 and N-linked glycosylation sites at Asn343 and Asn410. These features can affect apparent mass, but the supplied features do not establish a visible shift or a specific band position. An observed EPHA7 band size was not supplied.
Consider the listed splice variants before assigning the band: isoforms 3 and 5 lack large C-terminal regions, while isoforms 2 and 4 have smaller sequence changes. Check the antibody epitope against each sequence. The supplied features do not establish that any isoform produces a detectable band at a particular position.
Consider the listed splice variants before assigning the band: isoforms 3 and 5 lack large C-terminal regions, while isoforms 2 and 4 have smaller sequence changes. Check the antibody epitope against each sequence. The supplied features do not establish that any isoform produces a detectable band at a particular position.
Boster reagents
EPHA7 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-EphA7 (E477) Antibody
Cat # A03943
Real WB data
Anti-Ephrin type-A receptor 7 EphA7 Monoclonal Antibody
Cat # M03943
Two the supplier anti-EPHA7 antibodies have WB images. A03943 was tested on 3T3-L1, H9C2, A549, and HEK293T whole cell lysates at 1:500. M03943 was tested against truncated recombinant EPHA7 and transfected CHOK1 lysate; its image does not establish performance in untransfected samples.
Which to pick: Choose A03943 for a starting point with native cell lysates; its listed reactivity is human, mouse, and rat. M03943 lists human reactivity, but its WB image uses recombinant protein and transfected CHOK1 lysate.
Source: BosterBio EPHA7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15375.
- Human Protein Atlas. EPHA7 tissue expression.
- PMC5467824 — target-verified WB comparison
- PMC5085614 — target-verified WB comparison
- PMC4853368 — target-verified WB comparison