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ATG10 / Ubiquitin-like-conjugating enzyme ATG10 · Western blot design guide
Design a Western Blot for ATG10
Real validated ATG10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ATG10 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
ATG10 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 | ~25.3 kDa | |
| Observed band | ~28 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated ATG10 Western Blot Protocols
The M07803 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 % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M07803 · 1:50 (catalog M07803) |
| 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 ATG10 Western Blot Band Size?
ATG10 is predicted at 25.3 kDa and observed near 28 kDa; the cause of the difference is not established.
| Band near 28 kDa | Empirical ATG10 band in whole-cell lysates; confirm identity with controls |
| Band near 25.3 kDa | Near the UniProt predicted mass; confirm identity with controls |
| Several bands near the expected region | Isoforms 1 and 2 are annotated, but their migration is unknown |
| Single band without an isoform doublet | Isoforms 1 and 2 need not appear as distinct bands |
💡Expected ATG10 appearanceATG10 has a predicted mass of 25.3 kDa and an empirical band near 28 kDa; the reason for the difference is unestablished, so confirm band identity with ordinary controls.
| UniProt predicted mass | Sets a 25.3 kDa reference; the empirical band is near 28 kDa |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | Could affect band size, but no resolvable difference is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic ATG10 may be below detection in the tested sample | Check lysate quality, antibody performance, and a positive control |
| Band higher than expected | The observed ATG10 band is near 28 kDa versus a 25.3 kDa prediction; the cause is unknown | Compare with a positive control and verify band identity |
| Band lower than expected | Identity is uncertain; isoform migration is uncharacterized | Check band identity with an independent antibody or ATG10 depletion |
| Multiple bands | Two isoforms are annotated, but distinct migration is unproven | Compare bands with ATG10 depletion or an independent antibody |
| Weak or no signal | ATG10 may be below detection in the tested sample | Check loading, transfer, antibody performance, and a positive control |
Experiment evidence
Western blot images showing ATG10 band
M07803 · ATG10
M07803 · ATG10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ATG10 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ATG10 in Western blot, you can use adipose tissue.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Adipose tissue has medium expression, so the positive signal may be modest; ovary provides a not-detected negative tissue.
HPA tissue expression evidence for ATG10
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 | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced ATG10 Western Blot Tips
Deeper troubleshooting and optimisation questions for ATG10, answered from its protein features.
What should be checked if extra ATG10 bands appear?
Band shift · Compare their positions with the approximately 28 kDa observed band and check antibody recognition of both isoform sequences. Isoform 2 has a replacement at UniProt residues 73–125 and lacks residues 126–220. The listed interactions with MAP1LC3A, ATG5, ATG7 and IRGM do not, by themselves, identify an extra band.
How could ATG10 isoforms affect band interpretation?
Isoforms · ATG10 has two listed isoforms. In isoform 2, UniProt residues 73–125 are replaced by a different sequence and residues 126–220 are missing. This substantial sequence difference could affect apparent size or antibody recognition, but the features do not establish where either isoform runs on a blot.
An antibody recognizing a sequence within UniProt residues 126–220 could detect isoform 1 while missing isoform 2, because that region is absent from isoform 2. Check the antibody epitope against the listed sequences; these coordinates use UniProt numbering.
An antibody recognizing a sequence within UniProt residues 126–220 could detect isoform 1 while missing isoform 2, because that region is absent from isoform 2. Check the antibody epitope against the listed sequences; these coordinates use UniProt numbering.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does autophagy induction guarantee a stronger ATG10 band?
Induction · No. ATG10 is annotated in the autophagy and ubiquitin-like conjugation pathways, but the supplied features do not show that a particular treatment increases its abundance. Measure the signal under the chosen conditions rather than assuming induction.
How should transfer be checked for ATG10?
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 M07803 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 ATG10 bands be quantified?
Quantitation · Define the band being measured and use the same band assignment across samples. The approximately 28 kDa observation can guide that assignment, but the two listed isoforms make antibody recognition relevant. Verify which isoform the antibody detects before combining signals from different bands.
Why might ATG10 appear near 28 kDa instead of 25.3 kDa?
Interpretation · The observed band is approximately 28 kDa, while the predicted mass is 25.3 kDa. The supplied features do not establish the cause of this difference. Confirm band identity before attributing it to a modification or isoform.
UniProt places ATG10 in the cytoplasm. Include a cytoplasmic sample when assessing whether the antibody detects ATG10, and consider the sample fraction when interpreting a weak or absent band.
UniProt places ATG10 in the cytoplasm. Include a cytoplasmic sample when assessing whether the antibody detects ATG10, and consider the sample fraction when interpreting a weak or absent band.
Boster reagents
ATG10 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-Apg10 (Atg10) Rabbit Monoclonal Antibody
Cat # M07803
M07803 is a rabbit monoclonal anti-ATG10 antibody listed for human, mouse, and rat. Its supplied image documents immunoprecipitation at 1:50 with a Western blot readout at 1:3,000; direct lysate Western blot performance is not established by this caption.
Which to pick: M07803 is the only listed option. Consider it for the stated species, but note that its available Western blot image comes from an immunoprecipitation experiment, so the evidence does not show a direct lysate blot.
Source: BosterBio ATG10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H0Y0.
- Human Protein Atlas. ATG10 tissue expression.
- PMC10847611 — target-verified WB comparison
- PMC11481594 — target-verified WB comparison
- PMC3527592 — target-verified WB comparison