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- Table of Contents
SETD7 / Histone-lysine N-methyltransferase SETD7 · Western blot design guide
Design a Western Blot for SETD7
Source-linked SETD7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SETD7 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
SETD7 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 | ~40.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Lymph node (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SETD7 Western Blot Protocol Options
The M02793 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 | 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 | M02793; 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 SETD7 Western Blot Band Size?
SETD7 is predicted at 40.7 kDa; no empirical band is supplied, and its listed features do not demonstrate altered migration.
| Single band near 40.7 kDa | Consistent with SETD7, pending band-identity controls |
| Band near 40.7 kDa in a nuclear fraction | Consistent with SETD7's nuclear localization |
| Weak band in a cytoplasmic fraction | Consistent with SETD7's nuclear localization |
| Several bands at different sizes | The single annotated isoform does not explain distinct SETD7 bands |
💡Expected SETD7 appearanceUniProt predicts SETD7 at 40.7 kDa; no empirical band size is supplied, so confirm a candidate band near that size with band-identity controls.
| UniProt predicted mass of 40.7 kDa | Sets the reference size for full-length SETD7 |
| Sequence-derived mass of 40,721 Da | Corresponds to approximately 40.7 kDa |
| Predicted mass for the 366-residue sequence | Describes the full-length protein |
| Predicted rather than measured mass | Does not establish the apparent position on a blot |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear SETD7 may be poorly recovered | Check a nuclear extract and a positive-control lysate |
| Band higher than expected | The supplied features do not explain higher migration | Check markers and test whether SETD7 knockdown reduces the band |
| Band lower than expected | No signal peptide or propeptide cleavage is annotated | Check sample integrity and test whether SETD7 knockdown reduces the band |
| Multiple bands | Only one SETD7 isoform is annotated | Compare each band before and after SETD7 knockdown |
| Weak or no signal | The preparation may contain little nuclear SETD7 | Enrich the nuclear fraction and include a positive-control lysate |
Experiment evidence
Western blot images showing SETD7 band
M02793 · SETD7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SETD7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SETD7 in Western blot, you can use lymph node lysate, which has high HPA expression.
Positive control: Lymph node (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: A high-expression tissue and a not-detected tissue are available, though SETD7's nuclear location may favor nuclear extracts.
HPA tissue expression evidence for SETD7
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 |
|---|---|---|---|---|
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | 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 → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SETD7 Western Blot Tips
Deeper troubleshooting and optimisation questions for SETD7, answered from its protein features.
Where should the SETD7 band migrate?
Band shift · The calculated mass is 40.7 kDa, so examine the region near 41 kDa. No observed band position is supplied; the calculated mass alone cannot establish its apparent migration.
Could annotated isoforms explain multiple SETD7 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Do annotated modifications explain a shifted SETD7 band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. These annotations provide no basis for assigning a shift to a specific modification.
Does this guide establish induction of SETD7?
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 SETD7?
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 M02793 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 SETD7 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.
Which sample fraction is appropriate for detecting SETD7?
Interpretation · SETD7 is annotated in the nucleus and on chromosomes. Include nuclear material in the sample, and use the same extraction approach when comparing samples.
Because SETD7 is annotated in the nucleus and on chromosomes, use consistent nuclear extraction and loading across samples. Quantify the band selected near its calculated 40.7 kDa mass only after checking its specificity.
SETD7 interacts with IPF1/PDX-1, but that interaction does not establish that a higher-mass band contains SETD7. The supplied features provide no observed band position or modification that identifies an unexpected band; verify its identity experimentally.
Because SETD7 is annotated in the nucleus and on chromosomes, use consistent nuclear extraction and loading across samples. Quantify the band selected near its calculated 40.7 kDa mass only after checking its specificity.
SETD7 interacts with IPF1/PDX-1, but that interaction does not establish that a higher-mass band contains SETD7. The supplied features provide no observed band position or modification that identifies an unexpected band; verify its identity experimentally.
Boster reagents
SETD7 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-SETD7/Set7 Rabbit Monoclonal Antibody
Cat # M02793
The catalog reports one anti-SETD7 Western blot antibody, M02793, a rabbit monoclonal reported to react with human, mouse, and rat. Its WB image shows Jurkat and HeLa cell lysates; no other sample contexts are supplied.
Which to pick: M02793 is the only listed option and has a WB image using Jurkat and HeLa lysates. Check its reported reactivity against your sample species; the supplied image does not demonstrate every listed species.
Source: BosterBio SETD7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8WTS6.
- Human Protein Atlas. SETD7 tissue expression.
- PMC12914869 — target-verified WB comparison
- PMC10291335 — target-verified WB comparison
- PMC5706857 — target-verified WB comparison