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- Table of Contents
KEL / Kell blood group glycoprotein · Western blot design guide
Design a Western Blot for KEL
Source-linked KEL Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KEL 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
KEL 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 | ~82.8 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked KEL Western Blot Protocol Options
The A04711 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse-heart cells (catalog A04711) |
| 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 | A04711 · 1:1000 (catalog A04711) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A04711) |
| 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 KEL Western Blot Band Size?
KEL’s predicted polypeptide mass is 82.8 kDa; glycosylation and XK linkage may affect migration, but no empirical band size or visible shift is established.
| Band near 82.8 kDa | Compatible with the predicted KEL polypeptide mass; confirm identity with controls |
| Band above 82.8 kDa | N-linked glycans could affect migration, but a visible shift is not established |
| Higher band under nonreducing conditions | Could represent disulfide-linked KEL with XK; its band size is not supplied |
| Weak band in a soluble fraction | KEL is a membrane protein and may remain in the membrane fraction |
💡Expected KEL appearanceKEL has a predicted polypeptide mass of 82.8 kDa, but no empirical band size is supplied; use reduction, glycosidase treatment, and band-identity controls to assess any differing migration.
| Predicted KEL polypeptide mass | 82.8 kDa before any apparent migration effects |
| N-linked glycosylation at Asn94 | Could increase apparent size if occupied; no shift is established |
| N-linked glycosylation at Asn115 | Could increase apparent size if occupied; no shift is established |
| N-linked glycosylation at Asn191 in KEL2 antigen | Could increase apparent size if occupied; no shift is established |
| N-linked glycosylation at Asn345 and Asn627 | Could affect apparent size if occupied; no shift is established |
| Disulfide-linked heterodimer with XK | Could produce a higher band under nonreducing conditions; its size is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KEL may be poorly recovered from the membrane | Check membrane extraction and probe a membrane fraction |
| Band higher than expected | N-linked glycans or disulfide-linked XK may affect migration | Compare glycosidase-treated and fully reduced samples |
| Band lower than expected | Glycan occupancy may vary, or the band may be unrelated to KEL | Compare glycosidase-treated samples and verify band identity |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Compare with glycosidase-treated samples and verify band identity |
| Multiple bands | KEL monomer and disulfide-linked KEL–XK could coexist after incomplete reduction | Compare reducing and nonreducing lanes and verify band identity |
Experiment evidence
Western blot images showing KEL band
A04711 · KEL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KEL Western blot
🧪For positive controls for KEL in Western blot, you can use erythrocyte membranes, where UniProt places KEL; no HPA positive sample 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: Membrane-enriched samples are appropriate for KEL, but HPA data do not establish a tissue negative control.
HPA tissue expression evidence for KEL
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 KEL Western Blot Tips
Deeper troubleshooting and optimisation questions for KEL, answered from its protein features.
How should KEL 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 annotated KEL isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore do not support assigning separate bands to annotated isoforms. Check other feature-based possibilities, including glycosylation and the disulfide-linked XK heterodimer.
Which glycosylation sites matter when assessing a KEL band?
PTM · UniProt lists N-linked sites at Asn94, Asn115, Asn191, Asn345 and Asn627; Asn191 is annotated in the KEL2 antigen. These are UniProt coordinates. Record the band position before and after any glycosylation comparison rather than assuming a particular shift.
UniProt annotates phosphoserine at position 7. That site alone does not establish a visible band shift or identify an extra band. Use UniProt numbering when comparing the site with antibody or paper coordinates.
UniProt annotates phosphoserine at position 7. That site alone does not establish a visible band shift or identify an extra band. Use UniProt numbering when comparing the site with antibody or paper coordinates.
Does this guide establish induction of KEL?
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 KEL Western blot?
Transfer · KEL is an 82.8 kDa single-pass membrane protein. Choose transfer conditions that recover a protein of this size from the gel, then check the post-transfer gel and membrane for incomplete transfer. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04711 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 KEL Western blot signal be quantified?
Quantitation · KEL spans the erythrocyte membrane and is attached to the underlying cytoskeleton. Compare samples using consistent membrane preparation and loading, and measure bands within the detection system’s linear range. Define the band being measured before comparing intensities.
Should KEL migrate at its predicted 82.8 kDa?
Interpretation · 82.8 kDa is the predicted mass; no observed band position is supplied. KEL has five annotated N-linked glycosylation sites, but those annotations alone cannot establish a visible shift or explain a measured difference.
KEL forms a disulfide-linked heterodimer with XK and has six annotated disulfide bonds. Compare reducing and nonreducing samples when assessing a higher band. The five N-linked sites are also relevant to migration, but no observed band position is supplied, so these features cannot identify an unexpected band by themselves.
KEL forms a disulfide-linked heterodimer with XK and has six annotated disulfide bonds. Compare reducing and nonreducing samples when assessing a higher band. The five N-linked sites are also relevant to migration, but no observed band position is supplied, so these features cannot identify an unexpected band by themselves.
Boster reagents
KEL 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-CD238 KEL Antibody
Cat # A04711
A04711 is an anti-KEL (CD238) antibody listed for human and mouse reactivity. Its supplied Western blot image shows mouse-heart cells with primary antibody at 1:1000 and secondary antibody at 1:20000. No human WB example is supplied.
Which to pick: A04711 is the only listed option. It has a mouse-heart-cell WB image; human reactivity is listed, but no human WB example is provided.
Source: BosterBio KEL gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P23276.
- Human Protein Atlas. KEL tissue expression.
- PMC6004516 — target-verified WB comparison