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PEG10 / Retrotransposon-derived protein PEG10 · Western blot design guide
Design a Western Blot for PEG10
Real validated PEG10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PEG10 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
PEG10 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 | ~80.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Placenta (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Real Curated PEG10 Western Blot Protocols
The A03240 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A03240; 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 PEG10 Western Blot Band Size?
Full-length PEG10 is predicted at 80.2 kDa; isoforms, alternative initiation, and oligomerization could affect the pattern, but no empirical migration is supplied.
| Band near 80.2 kDa | Consistent with the predicted full-length PEG10 size; confirm identity with controls. |
| Band below 80.2 kDa | Could reflect alternative initiation or a shorter isoform; its identity is unconfirmed. |
| Several bands at different positions | Could reflect PEG10 isoforms 1–5; distinct migration has not been demonstrated. |
| High-molecular-weight material | Could reflect homooligomers if they persist during sample preparation. |
💡Expected PEG10 appearanceFull-length PEG10 has a predicted mass of 80.2 kDa; no empirical band size is supplied, so identify any candidate band with appropriate controls.
| Predicted full-length mass | Provides an 80.2 kDa reference, not a measured migration position. |
| Isoforms 1–5 | May differ in size; their individual masses and migration positions are not supplied. |
| Alternative initiation | May produce a shorter product, but no product size is supplied. |
| Homooligomerization | May produce higher-molecular-weight material if assemblies survive sample preparation; no band position is established. |
| Arginine methylation at residues 507, 598, and 611 | Its effect on apparent band size is not established. |
| Phosphoserine annotation at residue 611 | Its effect on apparent band size is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PEG10 can be released in extracellular vesicles, reducing the amount retained in the sampled cells. | Check the extracellular vesicle fraction alongside lysate and use a positive control. |
| Band higher than expected | PEG10 homooligomers may persist during sample preparation. | Compare denaturing sample conditions and confirm the signal with an independent PEG10 antibody. |
| Band lower than expected | Alternative initiation or a shorter isoform may account for a lower band. | Confirm identity with an antibody to another PEG10 region or a PEG10 depletion control. |
| Multiple bands | PEG10 has five annotated isoforms, but distinct band positions are unconfirmed. | Use PEG10 depletion and region-specific antibodies to identify which bands contain PEG10. |
| Weak or no signal | PEG10 may be distributed among cytoplasm, nucleus, and extracellular vesicles. | Check those fractions and verify antibody performance with a positive control. |
Experiment evidence
Western blot images showing PEG10 band
A03240 · PEG10
A03240-1 · PEG10
A03240-2 · PEG10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PEG10 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PEG10 in Western blot, you can use placenta tissue, which HPA reports as highly positive.
Positive control: Placenta (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: PEG10 can be released in extracellular vesicles, so signal may vary between cell lysate and extracellular vesicle samples.
HPA tissue expression evidence for PEG10
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 |
|---|---|---|---|---|
| Placenta | cytotrophoblasts | High | Protein (IHC) | HPA → |
| Testis | elongated or late spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Skin | cells in granular layer | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PEG10 Western Blot Tips
Deeper troubleshooting and optimisation questions for PEG10, answered from its protein features.
How should PEG10 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.
Which PEG10 isoforms could produce shorter bands?
Isoforms · Isoforms 2, 3, and 5 replace residues 320–325 and lack residues 326–708. Isoform 3 also has an altered N terminus, as do isoforms 4 and 5. Compare any band assignment with the isoform sequence recognized by the antibody.
Define whether the measurement targets one isoform or total detectable PEG10. Isoforms 2, 3, and 5 lack residues 326–708, so an antibody against that region would exclude them. Keep the same band-selection rule across samples and report which bands were quantified.
Define whether the measurement targets one isoform or total detectable PEG10. Isoforms 2, 3, and 5 lack residues 326–708, so an antibody against that region would exclude them. Keep the same band-selection rule across samples and report which bands were quantified.
Which PEG10 modifications matter when evaluating bands?
PTM · UniProt lists Omega-N-methylarginine at positions 507 and 598. Its supplied annotation for position 611 reads ‘Omega-N-methylarginine Phosphoserine Phosphoserine.’ These are UniProt coordinates; antibody or paper numbering may differ. Positions 507, 598, and 611 are absent from isoforms 2, 3, and 5. The annotations alone do not establish a visible shift.
Does this guide establish induction of PEG10?
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 PEG10 Western blot?
Transfer · The supplied features do not specify a transfer method. Validate transfer across the canonical protein’s predicted 80.2 kDa range and any shorter isoforms you intend to detect, since isoforms 2, 3, and 5 lack residues 326–708.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03240 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 PEG10 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might PEG10 bands differ from the predicted 80.2 kDa?
Interpretation · The 80.2 kDa prediction is for the canonical 708-residue sequence. Isoforms 2, 3, and 5 lack residues 326–708, so they may appear at lower masses. No observed band mass is supplied; the listed features alone cannot explain a specific difference between apparent and calculated mass.
An antibody recognizing residues 326–708 would not be expected to detect isoforms 2, 3, or 5, because that region is missing from them. An epitope within residues 320–325 also needs checking: KLPGPA becomes NSPAPL in those three isoforms.
First check whether the antibody epitope is present in the candidate isoform. Isoforms 2, 3, and 5 lack residues 326–708, while isoforms 3, 4, and 5 have altered N termini. PEG10 also has a listed homooligomerization feature. These features suggest possibilities but cannot identify an unexpected band without further evidence.
An antibody recognizing residues 326–708 would not be expected to detect isoforms 2, 3, or 5, because that region is missing from them. An epitope within residues 320–325 also needs checking: KLPGPA becomes NSPAPL in those three isoforms.
First check whether the antibody epitope is present in the candidate isoform. Isoforms 2, 3, and 5 lack residues 326–708, while isoforms 3, 4, and 5 have altered N termini. PEG10 also has a listed homooligomerization feature. These features suggest possibilities but cannot identify an unexpected band without further evidence.
Boster reagents
PEG10 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-PEG10 Monoclonal Antibody
Cat # A03240
Real WB data
Anti-PEG10 Monoclonal Antibody
Cat # A03240-1
Real WB data
Anti-PEG10 Antibody
Cat # A03240-2
Three the supplier anti-PEG10 antibodies have WB images: A03240 with recombinant PEG10 and transfected CHO-K1 lysate, A03240-1 with HeLa lysate, and A03240-2 with extracts from unspecified cell lines. These images document the stated test contexts, not every listed species.
Which to pick: For HeLa lysate, consider A03240-1. For recombinant or transfected PEG10, consider A03240. A03240-2 lists human, mouse, and rat reactivity and reports WB conditions, but its caption does not identify the cell lines tested. All three have WB images.
Source: BosterBio PEG10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86TG7.
- Human Protein Atlas. PEG10 tissue expression.
- PMC7212762 — target-verified WB comparison
- PMC13211853 — target-verified WB comparison
- PMC2800197 — target-verified WB comparison