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- Table of Contents
BOK / Bcl-2-related ovarian killer protein · Western blot design guide
Design a Western Blot for BOK
Real validated BOK Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BOK 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
BOK 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 | ~23.3 kDa | |
| Observed band | ~27 kDa | |
| Gel | 5–20% (catalog A05577-3) | |
| Negative control | Suggested KO / knockdown lysate |
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 | 2 isoform(s) |
Section 1
Real Curated BOK Western Blot Protocols
The A05577-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human HepG2, rat ovary (catalog A05577-3) |
| Gel % | 5–20% (catalog A05577-3) |
| Load | 30 ug; reducing conditions (catalog A05577-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05577-3) |
| Membrane | nitrocellulose membrane (catalog A05577-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05577-3) |
| Primary antibody | A05577-3 · 0.5 μg/mL (catalog A05577-3) |
| Primary incubation | overnight at 4°C (catalog A05577-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05577-3) |
| Secondary incubation | 1.5 hour at RT (catalog A05577-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05577-3) |
| Detection | ECL (catalog A05577-3) |
Section 2
What Is the Expected BOK Western Blot Band Size?
BOK is predicted at 23.3 kDa and observed at ~27 kDa on antibody QC blots; the cause of the difference is not established.
| Band at ~27 kDa | Empirical BOK band in reducing whole-cell and tissue lysates |
| Band near 23.3 kDa | Compatible with the predicted mass of BOK; identity requires confirmation |
| Band near twice the monomer mass | Could reflect the reported homodimer if it survives sample preparation |
| Multiple bands near the monomer region | Could reflect isoforms 1 and 2, whose migration has not been established |
💡Expected BOK appearanceBOK has a predicted mass of 23.3 kDa, while antibody QC detects a band at ~27 kDa under reducing conditions; confirm band identity with appropriate controls because the cause of the difference is not established.
| Predicted monomer mass | 23.3 kDa by sequence; the empirical BOK band is ~27 kDa |
| Homodimer formation | Could produce a species near twice the monomer mass if preserved during electrophoresis |
| Apoptosis-associated oligomerization | Could produce higher apparent species if oligomers persist during electrophoresis |
| Splice isoforms 1 and 2 | May differ in size, but their masses and band separation are not supplied |
| Phosphoserine at residue 7 | May affect migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated BOK may be poorly recovered | Check membrane extraction and compare with an enriched membrane fraction |
| Band higher than expected | The ~27 kDa empirical band exceeds the 23.3 kDa prediction; the cause is unknown | Compare with the QC band and verify identity using an independent antibody or BOK depletion |
| Band lower than expected | An uncharacterized isoform or protein breakdown could contribute | Compare fresh preparations and verify the band by BOK depletion |
| Multiple bands | Isoforms or retained BOK oligomers are possible, but band assignments are unproven | Compare reducing conditions and verify each band by BOK depletion |
| Weak or no signal | Recovery of membrane-associated BOK may be low | Check extraction, loading, and antibody performance with a positive sample |
Experiment evidence
Western blot images showing BOK band
A05577-3 · BOK
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BOK Western blot
🧪For positive controls for BOK in Western blot, you can use a BOK-positive sample validated independently, since no HPA positive tissue or cell is supplied.
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: BOK is membrane-associated, and the supplied HPA data provide no tissue-based controls; validate a positive sample and use knockdown or KO for a negative control.
HPA tissue expression evidence for BOK
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 BOK Western Blot Tips
Deeper troubleshooting and optimisation questions for BOK, answered from its protein features.
How should BOK 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 BOK isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–78 of isoform 1, so the two may produce different bands if both are detected. An antibody recognizing that missing region would not be expected to detect isoform 2.
Does BOK phosphorylation explain a band shift?
PTM · UniProt lists phosphoserine at position 7 in the canonical sequence. That position lies within the segment missing from isoform 2. Phosphorylation alone does not establish a visible shift or explain the approximately 27 kDa observed band; check which isoform and numbering convention an antibody or paper uses.
Could apoptotic stimulation change BOK band patterns?
Induction · UniProt states that apoptotic stimuli and BH3-only proteins promote BOK oligomerization. This supports considering oligomeric species when interpreting additional bands, but does not establish that stimulation increases BOK abundance or that oligomers remain visible after sample preparation.
What transfer method to use for BOK Western blot?
Transfer · UniProt places isoform 1 at mitochondrial and endoplasmic reticulum membranes and describes it as a single-pass membrane protein. Check transfer recovery for membrane-associated BOK using the post-transfer gel and membrane. These features do not specify one optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05577-3 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 BOK bands be quantified across samples?
Quantitation · Use the same band definition across samples and report whether quantitation includes one or both isoforms. Because isoform 1 is membrane-associated and BOK can oligomerize, changes in fraction preparation or which bands are counted could change the measured signal without establishing a change in total BOK abundance.
Why might BOK run near 27 kDa instead of 23.3 kDa?
Interpretation · BOK’s predicted mass is 23.3 kDa, while the supplied observed band is approximately 27 kDa. The listed features do not establish the cause of that difference. Compare the band with an appropriate molecular weight marker and assess it alongside other detected bands.
UniProt describes BOK homodimers and heterodimers by similarity, and oligomerization promoted by apoptotic stimuli and BH3-only proteins. Higher bands could be investigated as possible complexes, but band position alone cannot identify them. The listed phosphoserine at canonical position 7 does not establish a visible higher band.
UniProt describes BOK homodimers and heterodimers by similarity, and oligomerization promoted by apoptotic stimuli and BH3-only proteins. Higher bands could be investigated as possible complexes, but band position alone cannot identify them. The listed phosphoserine at canonical position 7 does not establish a visible higher band.
Boster reagents
BOK 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-BOK Antibody Picoband®
Cat # A05577-3
The catalog reports one anti-BOK antibody for Western blotting, A05577-3. Its blot image shows a band near 27 kDa, versus an expected 23 kDa, in the reported human cell and rat and mouse tissue lysates. Evidence beyond these samples is not supplied.
Which to pick: A05577-3 is the only listed option and has a Western blot image. The caption reports human MCF-7 and HepG2 cells, plus rat and mouse ovary and brain tissues; choose it with those tested contexts and the 27 kDa observed band in mind.
Source: BosterBio BOK gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UMX3.
- Human Protein Atlas. BOK tissue expression.
- PMC5689661 — target-verified WB comparison
- PMC12836499 — target-verified WB comparison