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KLRG1 / Killer cell lectin-like receptor subfamily G member 1 · Western blot design guide
Design a Western Blot for KLRG1
Real validated KLRG1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KLRG1 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
KLRG1 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 | ~21.8 kDa | |
| Observed band | ~38 kDa | |
| Gel | 10% (catalog M02084) | |
| Positive control | Tonsil (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Disulfide-linked | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated KLRG1 Western Blot Protocols
The M02084 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human Raji, human Hela (catalog M02084) |
| Gel % | 10% (catalog M02084) |
| Load | 30 ug; reducing conditions (catalog M02084) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02084) |
| Membrane | nitrocellulose membrane (catalog M02084) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02084) |
| Primary antibody | M02084 · 1:500 (catalog M02084) |
| Primary incubation | overnight at 4°C (catalog M02084) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M02084) |
| Secondary incubation | 1.5 hour at RT (catalog M02084) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02084) |
| Detection | ECL (catalog M02084) |
Section 2
What Is the Expected KLRG1 Western Blot Band Size?
KLRG1 is predicted at 21.8 kDa but shows an empirical band near 38 kDa under reducing conditions; the cause of the difference is not established.
| Band near 38 kDa | Empirical KLRG1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 21.8 kDa | Could correspond to the predicted monomer mass; identity requires confirmation |
| Higher band near twice the monomer mass | Could reflect disulfide-linked homodimer under non-reducing or incompletely reducing conditions |
| Multiple bands | Could reflect isoforms 1 and 2 or variable N-linked glycosylation; distinct migration is unproven |
💡Expected KLRG1 appearanceKLRG1 has a predicted mass of 21.8 kDa, while reducing whole-cell blots show a band near 38 kDa; the cause of this difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | 21.8 kDa is the sequence-based reference, not the observed migration |
| N-linked sites at Asn65, Asn97, Asn137 and Asn150 | Glycosylation could alter apparent mass; the sites alone do not establish a visible shift |
| Disulfide-linked homodimer | May produce a higher band under non-reducing or incompletely reducing conditions |
| Isoforms 1 and 2 | May differ in size, but their masses and migration difference are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated KLRG1 may be poorly represented in the preparation | Check membrane extraction and use a KLRG1-positive sample |
| Band higher than expected | Disulfide-linked homodimer may persist if reduction is incomplete; the 38 kDa band has no established cause | Compare reducing and non-reducing lanes and confirm identity with a second antibody or KLRG1 perturbation |
| Band lower than expected | The lower band may be a distinct species or nonspecific signal | Compare with a KLRG1-positive sample and verify identity by KLRG1 perturbation |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible at four annotated sites | Compare untreated and deglycosylated samples and confirm the signal is KLRG1 |
| Multiple bands | Isoforms, glycosylation or incompletely reduced homodimer may contribute | Compare reduction conditions and verify each band with KLRG1 perturbation |
| Weak or no signal | KLRG1 recovery from the membrane fraction may be low | Check membrane extraction and include a KLRG1-positive control |
Experiment evidence
Western blot images showing KLRG1 band
M02084 · KLRG1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KLRG1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KLRG1 in Western blot, you can use tonsil tissue lysate.
Positive control: Tonsil (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: KLRG1 is a membrane protein, so membrane enrichment may improve detection.
HPA tissue expression evidence for KLRG1
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 |
|---|---|---|---|---|
| Tonsil | non-germinal center cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KLRG1 Western Blot Tips
Deeper troubleshooting and optimisation questions for KLRG1, answered from its protein features.
How should KLRG1 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 KLRG1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, residues 187–195 of the canonical sequence (CPFADQALF) are replaced by VRL. Check which isoform your sample or antibody targets; the sequence difference alone does not establish separate visible bands.
Which KLRG1 glycosylation sites matter when interpreting band migration?
PTM · UniProt lists N-linked glycosylation at Asn 65, 97, 137, and 150. These are UniProt sequence coordinates; check the numbering convention before comparing them with an antibody or paper. The annotations do not establish a visible shift at any individual site.
Does this guide establish induction of KLRG1?
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 KLRG1?
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 M02084 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 KLRG1 bands be quantified consistently?
Quantitation · Define which band or bands you will measure and use the same choice across samples. KLRG1 can form monomers and disulfide-linked homodimers, so record whether samples were reduced when comparing intensities.
Why might KLRG1 appear near 38 kDa instead of 21.8 kDa?
Interpretation · The observed band is approximately 38 kDa, while the predicted sequence mass is 21.8 kDa. KLRG1 has four annotated N-linked glycosylation sites, but those features alone do not establish the cause or size of the difference.
KLRG1 forms monomers and disulfide-linked homodimers and has three annotated disulfide bonds. Compare reducing and nonreducing samples when assessing a higher band. Its position alone cannot confirm that the band is a dimer.
Consider its four N-linked glycosylation sites, two isoforms, and disulfide-linked homodimer formation. Compare reducing and nonreducing samples and record which isoform the antibody can recognize. These features suggest possibilities but cannot identify an unexpected band by position alone.
KLRG1 forms monomers and disulfide-linked homodimers and has three annotated disulfide bonds. Compare reducing and nonreducing samples when assessing a higher band. Its position alone cannot confirm that the band is a dimer.
Consider its four N-linked glycosylation sites, two isoforms, and disulfide-linked homodimer formation. Compare reducing and nonreducing samples and record which isoform the antibody can recognize. These features suggest possibilities but cannot identify an unexpected band by position alone.
Boster reagents
KLRG1 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-KLRG1 Rabbit Monoclonal Antibody
Cat # M02084
The catalog reports one anti-KLRG1 antibody for Western blot: M02084, a rabbit monoclonal with reported human, mouse, and rat reactivity. Its blot caption reports a band near 38 kDa, versus an expected 22 kDa; no additional validation evidence is supplied.
Which to pick: M02084 is the only listed option. Its WB image uses human cell lysates and rat and mouse spleen and lung lysates under reducing conditions, with the primary antibody at 1:500. Consider the reported band-size discrepancy when interpreting results.
Source: BosterBio KLRG1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96E93.
- Human Protein Atlas. KLRG1 tissue expression.
- PMC11251798 — target-verified WB comparison
- PMC10182695 — target-verified WB comparison
- PMC12044824 — target-verified WB comparison