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- Table of Contents
GPKOW / G-patch domain and KOW motifs-containing protein · Western blot design guide
Design a Western Blot for GPKOW
Source-linked GPKOW Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GPKOW WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
GPKOW 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 | ~52.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked GPKOW Western Blot Protocol Options
The A11892 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A11892) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A11892) |
| Primary antibody | A11892 · 1:3000 (catalog A11892) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A11892) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A11892) |
Section 2
What Is the Expected GPKOW Western Blot Band Size?
GPKOW is predicted at 52.2 kDa; its listed phosphorylation sites may affect migration, but no empirical band size or migration effect is established.
| Band near 52.2 kDa | Consistent with predicted GPKOW mass; confirm identity with controls |
| Weak band in a cytoplasmic fraction | Consistent with GPKOW's nuclear location |
| Doublet near 52.2 kDa | Could reflect listed phosphorylation sites, but this pattern is unverified |
| Band above 52.2 kDa | Altered migration is possible, but its cause is unestablished |
💡Expected GPKOW appearanceGPKOW has a predicted mass of 52.2 kDa; no empirical band size is supplied, so verify a candidate band near that size with antibody and fractionation controls.
| Predicted molecular mass | Places the protein sequence near 52.2 kDa |
| Phosphoserine at Ser27 by PKA | May affect migration; no visible size change is established |
| Phosphoserine at Ser35 | May affect migration; no visible size change is established |
| Phosphothreonine at Thr316 by PKA | May affect migration; no visible size change is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear GPKOW may be poorly recovered | Check nuclear extraction and protein loading |
| Band higher than expected | The cause of altered migration is unestablished | Verify identity with GPKOW depletion or a second antibody |
| Band lower than expected | Possible degradation or nonspecific binding | Use fresh lysate and verify with a second epitope antibody |
| Multiple bands | Possible phosphorylation states or nonspecific binding | Compare phosphatase-treated samples and verify band identity |
| Weak or no signal | Insufficient nuclear protein or antibody sensitivity | Check nuclear protein loading and a positive control |
Experiment evidence
Western blot images showing GPKOW band
A11892 · GPKOW
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GPKOW Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GPKOW in Western blot, you can use adrenal gland lysate, which HPA scores as High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: A liver negative control is available; GPKOW’s nuclear location may favor nuclear-enriched lysate.
HPA tissue expression evidence for GPKOW
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced GPKOW Western Blot Tips
Deeper troubleshooting and optimisation questions for GPKOW, answered from its protein features.
How should GPKOW 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.
Are alternative isoforms expected to produce separate GPKOW bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning a second band to an alternative GPKOW isoform.
Could GPKOW modifications explain a shifted band?
PTM · UniProt lists N-acetylalanine at position 2; phosphoserine at 27, 35, 42, 115, and 471; and phosphothreonine at 216, 316, and 473. PKA is specified for positions 27 and 316. These features alone do not establish a visible shift or explain any difference from 52.2 kDa. Positions use UniProt numbering, which may differ from paper or antibody numbering.
Does this guide establish induction of GPKOW?
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 GPKOW?
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 A11892 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 GPKOW bands be quantified?
Quantitation · Use a consistently defined candidate band across samples and report any additional bands separately. The 52.2 kDa predicted mass and nuclear localization can guide band selection, but the supplied features do not establish which observed band, if any, is GPKOW.
Where should GPKOW appear on a Western blot?
Interpretation · Use 52.2 kDa as the predicted mass when identifying a candidate band. No observed band position is supplied, so the actual migration cannot be specified.
UniProt assigns phosphorylation by PKA to serine 27 and threonine 316. If comparing phosphorylation states, track these sites using UniProt numbering and assess any band difference experimentally; their annotation does not predict a visible shift.
GPKOW is annotated in the nucleus and as a component of the minor spliceosome. A nuclear fraction is therefore a relevant sample to examine when checking a candidate GPKOW band.
Check each band's apparent mass against 52.2 kDa and whether it appears in nuclear samples. The record lists phosphorylation and N-terminal acetylation, but neither proves that a particular extra band is modified GPKOW. It supplies no alternative sequence or observed band position to support a more specific assignment.
UniProt assigns phosphorylation by PKA to serine 27 and threonine 316. If comparing phosphorylation states, track these sites using UniProt numbering and assess any band difference experimentally; their annotation does not predict a visible shift.
GPKOW is annotated in the nucleus and as a component of the minor spliceosome. A nuclear fraction is therefore a relevant sample to examine when checking a candidate GPKOW band.
Check each band's apparent mass against 52.2 kDa and whether it appears in nuclear samples. The record lists phosphorylation and N-terminal acetylation, but neither proves that a particular extra band is modified GPKOW. It supplies no alternative sequence or observed band position to support a more specific assignment.
Boster reagents
GPKOW 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-GPKOW Antibody
Cat # A11892
The catalog reports A11892, an anti-GPKOW antibody with stated Human, Mouse, and Rat reactivity. Its Western blot image caption reports extracts from various cell lines, 25 µg per lane, and a 1:3000 primary dilution; the cell lines are not identified.
Which to pick: A11892 is the only listed option. It has a Western blot image and stated Human, Mouse, and Rat reactivity; the caption does not identify the cell lines tested, so check suitability for your sample.
Source: BosterBio GPKOW gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92917.
- Human Protein Atlas. GPKOW tissue expression.
- PMC3179746 — target-verified WB comparison