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KAT2B / Histone acetyltransferase KAT2B · Western blot design guide
Design a Western Blot for KAT2B
Real validated KAT2B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KAT2B 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
KAT2B 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 | ~93 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated KAT2B Western Blot Protocols
The A02403-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cells (catalog A02403-1) |
| 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 | A02403-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A02403-1) |
| 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 KAT2B Western Blot Band Size?
KAT2B is predicted at 93 kDa; no empirical band size is supplied, and its listed features do not demonstrate altered migration.
| Band near 93 kDa | Consistent with predicted full-length KAT2B; confirm its identity |
| Near-93 kDa band enriched in nuclear fraction | Consistent with mainly nuclear KAT2B |
| Near-93 kDa band in cytoplasmic fraction | Consistent with reported cytoplasmic KAT2B localization |
| No band near 93 kDa in nuclear fraction | KAT2B or its acetyl-K428 state may be below detection |
💡Expected KAT2B appearanceUniProt predicts 93 kDa for full-length KAT2B; no empirical band size is supplied, and the lead antibody recognizes acetyl-K428, so confirm candidate bands with identity and acetylation-state controls.
| UniProt predicted mass | Places full-length KAT2B near 93 kDa |
| Predicted full-length mass in lysate | Provides a 93 kDa reference for candidate bands |
| Predicted full-length mass in nuclear fraction | Provides the same 93 kDa reference in KAT2B's main compartment |
| Predicted full-length mass in cytoplasmic fraction | Provides the same 93 kDa reference where KAT2B is also reported |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KAT2B or its acetyl-K428 state may be below detection | Check a nuclear fraction and compare with a total-KAT2B antibody |
| Band higher than expected | The supplied features do not establish a size-raising modification | Verify identity with KAT2B depletion and a total-KAT2B antibody |
| Band lower than expected | The supplied features do not establish a cleavage product | Check identity with KAT2B depletion and another KAT2B antibody |
| Multiple bands | No distinct isoforms or processing products are listed | Identify KAT2B-dependent bands by depletion |
| Weak or no signal | The lead antibody recognizes acetyl-K428, a specific KAT2B state | Check a positive control and compare with total KAT2B |
Experiment evidence
Western blot images showing KAT2B band
A02403-1 · KAT2B
A02403 · KAT2B
A02403 · KAT2B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KAT2B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KAT2B in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for KAT2B
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 |
|---|---|---|---|---|
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | High | Protein (IHC) | HPA → |
| Skin | melanocytes | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Low | Protein (IHC) | HPA → |
| Breast | glandular cells | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced KAT2B Western Blot Tips
Deeper troubleshooting and optimisation questions for KAT2B, answered from its protein features.
Where should the KAT2B band appear?
Band shift · The supplied sequence is 832 amino acids with a predicted mass of 93 kDa. Use 93 kDa as a reference when identifying a candidate band; no observed band position is supplied.
Could KAT2B isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands.
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 this guide establish induction of KAT2B?
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 KAT2B Western blot?
Transfer · KAT2B is predicted to be 93 kDa. Check transfer efficiency near 93 kDa when selecting and optimizing a transfer method; the supplied features do not specify one method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02403-1 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 KAT2B 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.
Does a band above 93 kDa indicate modified KAT2B?
Interpretation · The supplied features do not establish a visible modification-dependent shift or an observed-versus-predicted mass difference. A band above 93 kDa alone cannot establish that KAT2B is modified.
KAT2B mainly localizes to the nucleus, so a nuclear fraction is a sensible starting point. It is also reported in the cytoplasm and at the centrosome; a weak nuclear signal alone does not establish absence from the sample.
Compare matched sample fractions and use a loading reference appropriate to that fraction. KAT2B is mainly nuclear but also has reported cytoplasmic and centrosomal localization, so differences between fractions need not reflect a change in total abundance.
The supplied interaction annotation says the unsumoylated form interacts with NR2C1. It gives no modification coordinate or evidence of a visible band difference, so band position alone cannot identify that form.
KAT2B mainly localizes to the nucleus, so a nuclear fraction is a sensible starting point. It is also reported in the cytoplasm and at the centrosome; a weak nuclear signal alone does not establish absence from the sample.
Compare matched sample fractions and use a loading reference appropriate to that fraction. KAT2B is mainly nuclear but also has reported cytoplasmic and centrosomal localization, so differences between fractions need not reflect a change in total abundance.
The supplied interaction annotation says the unsumoylated form interacts with NR2C1. It gives no modification coordinate or evidence of a visible band difference, so band position alone cannot identify that form.
Boster reagents
KAT2B 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-Acetyl-PCAF (K428) KAT2B Antibody
Cat # A02403-1
Real WB data
Anti-PCAF KAT2B Antibody
Cat # A02403
Two catalogued anti-KAT2B antibodies have WB images: A02403-1 shows acetyl-PCAF (K428) in HepG2 cells, while A02403 shows PCAF in nuclear extracts from various cells. The supplied captions do not establish performance across every listed species or a comparison between antibodies.
Which to pick: Choose A02403-1 for the acetyl-K428 form; its WB example uses HepG2 cells, and its listed reactivity is human and mouse. Choose A02403 for PCAF, including rat samples by listed reactivity; its WB example uses nuclear extracts from unspecified cells.
Source: BosterBio KAT2B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92831.
- Human Protein Atlas. KAT2B tissue expression.
- PMC8806817 — target-verified WB comparison
- PMC11772695 — target-verified WB comparison
- PMC12291418 — target-verified WB comparison