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- Table of Contents
SUZ12 / Polycomb protein SUZ12 · Western blot design guide
Design a Western Blot for SUZ12
Real validated SUZ12 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SUZ12 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
SUZ12 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 | ~83.1 kDa | |
| Observed band | Approximately 83 and 90 kDa | |
| Gel | 10% (catalog A00583-1) | |
| Positive control | Fallopian tube (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 | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SUZ12 Western Blot Protocols
The A00583-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Hela, human HepG2 (catalog A00583-1) |
| Gel % | 10% (catalog A00583-1) |
| Load | 30 ug; reducing conditions (catalog A00583-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00583-1) |
| Membrane | nitrocellulose membrane (catalog A00583-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00583-1) |
| Primary antibody | A00583-1 · 0.5 μg/mL (catalog A00583-1) |
| Primary incubation | overnight at 4°C (catalog A00583-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00583-1) |
| Secondary incubation | 1.5 hour at RT (catalog A00583-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00583-1) |
| Detection | ECL (catalog A00583-1) |
Section 2
What Is the Expected SUZ12 Western Blot Band Size?
SUZ12 is predicted at 83.1 kDa; bands near 83 and 90 kDa are observed, but the cause of their difference is not established.
| Band near 83 kDa | Matches the 83.1 kDa predicted mass and an observed SUZ12 band. |
| Band near 90 kDa | Matches a second observed SUZ12 band; its higher migration position has no established cause here. |
| Bands near 83 and 90 kDa | Both positions have been reported for SUZ12, but their relationship is unestablished. |
| Weak band in whole-cell lysate | SUZ12 localizes to the nucleus and chromatin; nuclear enrichment may improve detection. |
💡Expected SUZ12 appearanceSUZ12 has a predicted mass of 83.1 kDa and observed bands near 83 and 90 kDa; the difference is unexplained, so confirm band identity with appropriate antibody and SUZ12 depletion controls.
| Predicted mass of 83.1 kDa | Provides the full-length reference; an observed band is near 83 kDa. |
| Phosphoserine at Ser20 | Its effect on apparent band size is not established. |
| Phosphoserines at Ser541 and Ser546 | Their effect on apparent band size is not established. |
| Phosphoserines at Ser583 and Ser726 | Their effect on apparent band size is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear, chromatin-associated SUZ12 may be poorly recovered. | Check a nuclear-enriched extract and verify extraction with a nuclear marker. |
| Band higher than expected | A band near 90 kDa is reported, but its shift is unexplained. | Compare with a validated SUZ12-positive sample and test whether SUZ12 depletion removes the band. |
| Band lower than expected | The supplied features do not establish a smaller SUZ12 product. | Check band identity with SUZ12 depletion and another antibody epitope. |
| Multiple bands | Bands near 83 and 90 kDa are reported; no distinct isoforms are documented here. | Test each band with SUZ12 depletion and compare antibody epitopes. |
| Weak or no signal | Nuclear localization may limit the amount recovered in the tested sample. | Assess nuclear extraction, sample loading, and transfer. |
Experiment evidence
Western blot images showing SUZ12 band
A00583-1 · SUZ12
A00583-1 · SUZ12
A00583 · SUZ12
M00583 · SUZ12
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SUZ12 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SUZ12 in Western blot, you can use fallopian tube tissue, where HPA reports high expression.
Positive control: Fallopian tube (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: SUZ12 is chromatin-associated, so nuclear-enriched lysates may improve detection.
HPA tissue expression evidence for SUZ12
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 |
|---|---|---|---|---|
| Fallopian tube | ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | cells in tubules | 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SUZ12 Western Blot Tips
Deeper troubleshooting and optimisation questions for SUZ12, answered from its protein features.
How should SUZ12 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.
Could the reported SUZ12 bands represent listed isoforms?
Isoforms · Only one isoform is supplied, with no alternative sequence. The 83 and 90 kDa bands therefore cannot be assigned to separate listed isoforms from this evidence.
Which SUZ12 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserines at positions 20, 541, 546, 583, and 726. These are UniProt sequence coordinates, which may differ from paper or antibody numbering. Their presence does not establish that phosphorylation causes a detectable band shift.
Does this guide establish induction of SUZ12?
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 SUZ12?
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 A00583-1 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 SUZ12 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 SUZ12 appear near 83 and 90 kDa?
Interpretation · UniProt predicts 83.1 kDa, close to the reported 83 kDa band. The supplied features do not establish why a band appears near 90 kDa. Verify both bands before assigning them to SUZ12; the listed modifications alone do not demonstrate a visible shift.
SUZ12 is nuclear and localizes to chromatin as part of PRC2. Include a nuclear or chromatin-containing fraction when preparing samples, and account for the fraction used when comparing results.
Compare equivalent sample fractions and normalize within the same fraction. SUZ12 localizes to nuclear chromatin, so differences in nuclear or chromatin recovery could affect measured band intensity independently of SUZ12 abundance.
Check whether a band is reproducible and specific before identifying it as SUZ12. The supplied record predicts 83.1 kDa and lists five phosphoserines, but does not establish the identity or cause of an additional band.
SUZ12 is nuclear and localizes to chromatin as part of PRC2. Include a nuclear or chromatin-containing fraction when preparing samples, and account for the fraction used when comparing results.
Compare equivalent sample fractions and normalize within the same fraction. SUZ12 localizes to nuclear chromatin, so differences in nuclear or chromatin recovery could affect measured band intensity independently of SUZ12 abundance.
Check whether a band is reproducible and specific before identifying it as SUZ12. The supplied record predicts 83.1 kDa and lists five phosphoserines, but does not establish the identity or cause of an additional band.
Boster reagents
SUZ12 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-SUZ12 Antibody Picoband®
Cat # A00583-1
Real WB data
Anti-Polycomb protein SUZ12 SUZ12 Antibody
Cat # A00583
Real WB data
Anti-SUZ12 Rabbit Monoclonal Antibody
Cat # M00583
All three listed anti-SUZ12 antibodies have WB images. A00583-1 shows an approximately 83 kDa band in human, rat, and mouse cell lysates; A00583 shows human liver lysate; M00583 shows HeLa and SW480 lysates. These images document the tested samples only.
Which to pick: For human liver tissue, consider A00583, which has a liver WB image. For human, mouse, or rat cell lysates, A00583-1 has imaged examples. M00583 lists reactivity with all three species, but its WB image shows only HeLa and SW480 lysates.
Source: BosterBio SUZ12 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15022.
- Human Protein Atlas. SUZ12 tissue expression.
- PMC12482123 — target-verified WB comparison
- PMC12143176 — target-verified WB comparison
- PMC5175387 — target-verified WB comparison