This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
KDM4B / Lysine-specific demethylase 4B · Western blot design guide
Design a Western Blot for KDM4B
Real validated KDM4B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KDM4B 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
KDM4B 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 | ~121.9 kDa | |
| Observed band | ~150 kDa | |
| Gel | 8% (catalog M04127) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated KDM4B Western Blot Protocols
The M04127 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SW480, human 293T, human K562 (catalog M04127) |
| Gel % | 8% (catalog M04127) |
| Load | 30 ug; reducing conditions (catalog M04127) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04127) |
| Membrane | nitrocellulose membrane (catalog M04127) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04127) |
| Primary antibody | M04127 · 1:500 (catalog M04127) |
| Primary incubation | overnight at 4°C (catalog M04127) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M04127) |
| Secondary incubation | 1.5 hour at RT (catalog M04127) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04127) |
| Detection | ECL (catalog M04127) |
Section 2
What Is the Expected KDM4B Western Blot Band Size?
KDM4B is predicted at 121.9 kDa and observed near 150 kDa on reducing Western blots; the cause of the difference is not established.
| Band near 150 kDa | Empirical KDM4B band in reducing whole-cell lysates; identity still requires antibody controls |
| Band near 122 kDa | Near the 121.9 kDa UniProt predicted mass; identity requires confirmation |
| Multiple bands | Could reflect isoforms 1 and 2, but distinct migration is not established |
| Stronger band in a nuclear fraction | Consistent with KDM4B's nuclear location |
💡Expected KDM4B appearanceUniProt predicts 121.9 kDa, while antibody QC reports a band near 150 kDa in reducing lysates; the cause of the difference is unestablished, so confirm identity with antibody controls.
| UniProt predicted mass | Sets a sequence-based reference of 121.9 kDa; the empirical band is near 150 kDa |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | Could alter apparent size, but distinct band positions are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear KDM4B may be diluted in whole-cell lysate | Check a nuclear-enriched fraction and a positive-control lysate |
| Band higher than expected | The reported 150 kDa band exceeds the 121.9 kDa prediction for an undetermined reason | Compare with the reported band and confirm identity using an independent antibody or KDM4B depletion |
| Band lower than expected | A lower band could be a splice isoform or a fragment; its identity is unconfirmed | Use an independent epitope or KDM4B depletion to identify it |
| Multiple bands | Isoforms 1 and 2 are listed, but their band positions are unknown | Check which bands respond to KDM4B depletion |
| Weak or no signal | Nuclear localization may reduce signal in a whole-cell sample | Enrich the nuclear fraction and include a positive-control lysate |
Experiment evidence
Western blot images showing KDM4B band
M04127 · KDM4B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KDM4B Western blot
🧪For positive controls for KDM4B in Western blot, you can use no HPA-supported tissue or cell sample from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no expression data here, so a positive sample and a clean negative tissue cannot be identified.
HPA tissue expression evidence for KDM4B
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced KDM4B Western Blot Tips
Deeper troubleshooting and optimisation questions for KDM4B, answered from its protein features.
How should KDM4B 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.
How could isoform 2 affect the KDM4B band pattern?
Isoforms · Isoform 2 replaces residues 373–448 and lacks residues 449–1096, so it could produce a substantially smaller band than canonical isoform 1. A lower band should not be assigned to isoform 2 by size alone; check whether the antibody recognizes a region retained in that isoform.
Residues 1–372 are shared by both listed isoforms. An antibody targeting that region could detect both. An epitope within canonical residues 449–1096 would be absent from isoform 2, while residues 373–448 differ between the isoforms.
Residues 1–372 are shared by both listed isoforms. An antibody targeting that region could detect both. An epitope within canonical residues 449–1096 would be absent from isoform 2, while residues 373–448 differ between the isoforms.
Which listed modifications should guide KDM4B band interpretation?
PTM · UniProt lists phosphoserine 566, N6-acetyllysine 602, and phosphothreonine 1065. These are UniProt canonical-sequence coordinates, which may differ from paper or antibody numbering. All three positions fall within the region missing from isoform 2. Their presence alone does not demonstrate a visible band shift.
Does this guide establish induction of KDM4B?
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 KDM4B Western blot?
Transfer · The canonical predicted mass is 121.9 kDa, and the supplied observed band is near 150 kDa. Use transfer conditions validated for proteins in that size range, then check membrane transfer and residual protein in the gel. The supplied features do not identify one specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M04127 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 KDM4B bands be quantified?
Quantitation · KDM4B is annotated in the nucleus. Compare samples prepared with the same fractionation method and quantify the same resolved band across lanes. If multiple bands appear, report them separately unless their identities are established; the two listed isoforms differ substantially in sequence.
Why does KDM4B appear near 150 kDa instead of 121.9 kDa?
Interpretation · The canonical sequence has a predicted mass of 121.9 kDa, while the supplied observed band is near 150 kDa. The listed features do not establish the cause of that difference. Confirm band identity with an antibody recognizing a defined KDM4B region; do not assign the difference to a modification solely from its presence.
Consider isoform 2 because it lacks canonical residues 449–1096, but verify that the antibody epitope is retained before assigning the band. The supplied features list no signal peptide or propeptide and no glycosylation sites, so they provide no basis for attributing a lower band to those features.
Consider isoform 2 because it lacks canonical residues 449–1096, but verify that the antibody epitope is retained before assigning the band. The supplied features list no signal peptide or propeptide and no glycosylation sites, so they provide no basis for attributing a lower band to those features.
Boster reagents
KDM4B 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-KDM4B / JMJD2B Rabbit Monoclonal Antibody
Cat # M04127
The catalog reports one anti-KDM4B Western blot antibody, M04127, with stated human, mouse, and rat reactivity. Its WB image shows bands in the specified cell and tissue lysates; the reported band is approximately 150 kDa versus an expected 122 kDa.
Which to pick: M04127 is the only listed option. Its WB example includes human SW480, 293T, K562, and Jurkat cells and rat and mouse testis and brain tissue, using a 1:500 primary dilution. Consider the reported band-size discrepancy when interpreting results.
Source: BosterBio KDM4B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O94953.
- Human Protein Atlas. KDM4B tissue expression.
- PMC13249574 — target-verified WB comparison
- PMC13419031 — target-verified WB comparison
- PMC10038057 — target-verified WB comparison