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- Table of Contents
OGN / Mimecan · Western blot design guide
Design a Western Blot for OGN
Source-linked OGN Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OGN 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
OGN 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 | ~33.9 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 + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked OGN Western Blot Protocol Options
The A07061 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | L929 cells (catalog A07061) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A07061; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A07061) |
| 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 OGN Western Blot Band Size?
OGN has a predicted 33.9 kDa precursor; signal-peptide cleavage and glycosylation could affect migration, but no measured band or visible shift is supplied.
| Band near 33.9 kDa | Consistent with the predicted precursor mass; identity requires confirmation |
| Band above 33.9 kDa | Glycosylation at listed sites could alter migration; the shift is unverified |
| Band below 33.9 kDa | Could reflect removal of the 1–20 signal peptide; its migration is unverified |
| Little or no band in whole-cell lysate | OGN is secreted into the extracellular space and matrix |
💡Expected OGN appearanceOGN has a predicted precursor mass of 33.9 kDa, but signal-peptide cleavage and glycosylation may affect migration; no empirical band size is supplied, so confirm identity with appropriate controls.
| Predicted precursor mass | Provides a 33.9 kDa sequence-based reference, not a measured band |
| Signal peptide at residues 1–20 | Cleavage may yield a smaller mature protein; its apparent size is unknown |
| O-linked GalNAc site at Thr80 | May affect migration; a visible shift is not established |
| N-linked keratan sulfate site at Asn88 | May affect apparent size; the magnitude is unknown |
| N-linked keratan sulfate site at Asn214 | May affect apparent size; the magnitude is unknown |
| N-linked keratan sulfate site at Asn258 | May affect apparent size; the magnitude is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | OGN is secreted into the extracellular space and matrix | Check conditioned medium or extracellular matrix fractions alongside lysate |
| Band higher than expected | Glycosylation could alter migration | Compare untreated and suitably deglycosylated samples with an OGN identity control |
| Band lower than expected | Signal-peptide cleavage may reduce size | Compare precursor and mature protein recognition with a validated OGN control |
| Broad smear instead of sharp band | Variable glycosylation is possible but unconfirmed | Compare glycan-treated and untreated samples and confirm OGN identity |
| Multiple bands | Different processing or glycosylation states are possible | Check which bands track with an OGN-specific control and glycan treatment |
| Weak or no signal | Secreted OGN may be scarce in the sampled fraction | Check extracellular samples and verify antibody performance with a positive control |
Experiment evidence
Western blot images showing OGN band
A07061 · OGN
A07061 · OGN
A07061-2 · OGN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OGN Western blot
🧪For positive controls for OGN in Western blot, you can use an HPA-IHC candidate positive sample once identified; none is supplied here.
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 such as Ponceau alongside the samples.
⚠️Feasibility: Because OGN is secreted into the extracellular matrix, whole-cell lysates may give a weak signal.
HPA tissue expression evidence for OGN
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 OGN Western Blot Tips
Deeper troubleshooting and optimisation questions for OGN, answered from its protein features.
How should OGN 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.
Can annotated OGN isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Assess band identity alongside the annotated signal peptide, glycosylation sites and your sample context.
Which OGN glycosylation sites should I consider when interpreting bands?
PTM · UniProt annotates O-linked glycosylation at Thr80 and N-linked glycosylation associated with keratan sulfate at Asn88, Asn214 and Asn258. These are UniProt sequence coordinates; check the numbering convention before comparing them with an antibody or paper. The annotations alone do not show which sites are occupied in your sample.
Does this guide establish induction of OGN?
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 OGN?
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 A07061 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 I quantify OGN if several bands appear?
Quantitation · Define which band or band group you will measure and apply that choice consistently across samples. Report its apparent position: the record supplies a 33.9 kDa full-sequence prediction, four glycosylation sites and a signal peptide, but no empirical band position to identify a band by size alone.
Why might OGN migrate differently from its predicted 33.9 kDa?
Interpretation · The 33.9 kDa prediction is for the full 298-residue sequence. OGN has a signal peptide at residues 1–20 and four annotated glycosylation sites. These features affect how the predicted mass should be interpreted, but they do not establish an apparent band size or prove a visible shift; no observed band is supplied.
UniProt places the signal peptide at residues 1–20. Account for this region when comparing a secreted OGN band with the 33.9 kDa full-sequence prediction. The supplied features do not give an observed apparent mass.
OGN is annotated as secreted and located in the extracellular space and extracellular matrix. Consider those compartments when choosing material for the blot and interpreting a weak signal in a cell-only sample.
Check band identity before assigning a cause. OGN has a signal peptide at residues 1–20, four annotated glycosylation sites and one annotated disulfide bond. These features may inform follow-up comparisons, but their presence alone does not explain a particular unexpected band or demonstrate a visible shift.
UniProt places the signal peptide at residues 1–20. Account for this region when comparing a secreted OGN band with the 33.9 kDa full-sequence prediction. The supplied features do not give an observed apparent mass.
OGN is annotated as secreted and located in the extracellular space and extracellular matrix. Consider those compartments when choosing material for the blot and interpreting a weak signal in a cell-only sample.
Check band identity before assigning a cause. OGN has a signal peptide at residues 1–20, four annotated glycosylation sites and one annotated disulfide bond. These features may inform follow-up comparisons, but their presence alone does not explain a particular unexpected band or demonstrate a visible shift.
Boster reagents
OGN 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-Osteoglycin OGN Antibody
Cat # A07061
Real WB data
Anti-Mimecan OGN Antibody
Cat # A07061-2
Both listed anti-OGN antibodies have Western blot images. A07061 is shown with L929 cells and a secondary antibody diluted 1:20,000. A07061-2 is shown with extracts from various cell lines. No publication evidence is supplied.
Which to pick: For rat samples, choose A07061-2, the only one listed with rat reactivity. Both list human and mouse reactivity; A07061 provides a named L929 cell example, while A07061-2’s caption does not name the cell lines.
Source: BosterBio OGN gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P20774.
- Human Protein Atlas. OGN tissue expression.
- PMC5697837 — target-verified WB comparison
- PMC5833032 — target-verified WB comparison
- PMC6900314 — target-verified WB comparison