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KLRD1 / Natural killer cells antigen CD94 · Western blot design guide
Design a Western Blot for KLRD1
Source-linked KLRD1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KLRD1 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
KLRD1 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 | ~20.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Disulfide-linked | |
| Caveat | Reducing-condition controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked KLRD1 Western Blot Protocol Options
The A03257-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03257-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| 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 KLRD1 Western Blot Band Size?
KLRD1 has a predicted monomer mass of 20.5 kDa; glycosylation, isoforms, and disulfide-bonded heterodimers could affect migration, but no empirical band is supplied.
| Band near 20.5 kDa | Consistent with the predicted KLRD1 monomer; confirm identity with controls |
| Band above 20.5 kDa | Could reflect N-linked glycosylation at Asn83 or Asn132; a visible shift is unproven |
| Higher band under non-reducing conditions | Could reflect a disulfide-bonded KLRD1 heterodimer with an NKG2 family member |
| Several bands at different positions | Could reflect isoforms 1, 2, and 3, though distinct migration is unproven |
| Weak band in soluble lysate | KLRD1 is a single-pass cell-membrane protein that may remain in the membrane fraction |
💡Expected KLRD1 appearanceThe UniProt-predicted mass is 20.5 kDa; glycosylation, isoforms, and disulfide-bonded heterodimers could alter migration, but no empirical band size is supplied, so confirm band identity with appropriate controls.
| UniProt-predicted monomer mass | Provides a 20.5 kDa sequence-based reference, not a measured band |
| N-linked glycosylation at Asn83 | Could increase apparent size; the size of any shift is unknown |
| N-linked glycosylation at Asn132 | Could increase apparent size; the size of any shift is unknown |
| Disulfide-bonded heterodimer with KLRC1 or KLRC2 | Could produce a higher band under non-reducing conditions; its mass is unspecified |
| Isoforms 1, 2, and 3 | Could differ in size, but their masses and migration differences are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated KLRD1 may be poorly recovered in the sampled lysate | Check a membrane-enriched fraction and a known positive sample |
| Band higher than expected | N-linked glycosylation or a disulfide-bonded heterodimer may contribute | Compare deglycosylated and fully reduced samples with the untreated sample |
| Band lower than expected | Isoform-dependent migration or protein breakdown is possible but unconfirmed | Check sample integrity and verify the band with an independent antibody |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible but unproven | Compare paired untreated and deglycosylated samples |
| Multiple bands | Isoforms, glycosylation, or incomplete reduction may contribute | Compare reduced and deglycosylated samples and verify candidate bands with an independent antibody |
| Weak or no signal | Recovery of this cell-membrane protein may be low | Check membrane extraction and include a known positive sample |
Experiment evidence
Western blot images showing KLRD1 band
A03257-1 · KLRD1
A03257 · KLRD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KLRD1 Western blot
🧪For positive controls for KLRD1 in Western blot, you can use no HPA-supported sample because no positive tissue or cell data were 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: HPA provides no expression data to validate tissue controls for this membrane protein.
HPA tissue expression evidence for KLRD1
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 KLRD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for KLRD1, answered from its protein features.
How should KLRD1 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 KLRD1 isoforms produce different bands?
Isoforms · Three isoforms are listed. Relative to the canonical sequence, isoform 3 replaces residues 1–34 with MAA, while isoform 2 changes residue 105 from L to LQ. These sequence differences may affect migration or antibody recognition. Check which isoforms the antibody can detect before assigning bands.
How can glycosylation be assessed on a KLRD1 blot?
PTM · Compare matched samples before and after N-glycan removal. UniProt annotates N-linked sites at positions 83 and 132 in the supplied numbering. A mobility change would support glycosylation of the detected species; the annotations alone do not predict the size of a change.
Does this guide establish induction of KLRD1?
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 KLRD1 Western blot?
Transfer · KLRD1 is a 20.5 kDa, single-pass membrane protein. Use transfer conditions that retain proteins near 20 kDa, and check the membrane and post-transfer gel for recovery. The supplied features do not establish one optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03257-1 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 KLRD1 bands be quantified?
Quantitation · Quantify a consistently identified band under the same sample preparation and reduction conditions across lanes. KLRD1 has three isoforms, two annotated N-linked glycosylation sites, and can form disulfide-bonded heterodimers, so establish which species each band represents before combining their signals.
Why might KLRD1 migrate differently from its predicted 20.5 kDa mass?
Interpretation · KLRD1 has two annotated N-linked glycosylation sites, at UniProt positions 83 and 132. Glycosylation could affect migration, but these features alone do not establish a visible shift or explain any particular band. No observed band mass was supplied.
KLRD1 can form disulfide-bonded heterodimers with KLRC1 or KLRC2. Compare reducing and nonreducing samples when evaluating a higher-mass band. The five annotated disulfide bonds also make sample preparation relevant, but band position alone cannot identify a heterodimer.
KLRD1 can form disulfide-bonded heterodimers with KLRC1 or KLRC2. Compare reducing and nonreducing samples when evaluating a higher-mass band. The five annotated disulfide bonds also make sample preparation relevant, but band position alone cannot identify a heterodimer.
Boster reagents
KLRD1 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-CD94 KLRD1 Monoclonal Antibody
Cat # A03257-1
Real WB data
Anti-KLRD1 Antibody
Cat # A03257
Both listed antibodies have WB images. A03257 was shown with mouse brain extracts at 1:3000; A03257-1 was shown against recombinant protein. These examples support the reported test contexts, not broad validation across tissues or species.
Which to pick: For mouse samples, A03257 has a mouse brain WB example and lists Human, Mouse, and Rat reactivity. A03257-1 lists Human reactivity, but its WB image uses recombinant protein. Choose based on your sample and the evidence available for it.
Source: BosterBio KLRD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13241.
- Human Protein Atlas. KLRD1 tissue expression.
- PMC12495723 — target-verified WB comparison