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- Table of Contents
COL10A1 / Collagen alpha-1(X) chain · Western blot design guide
Design a Western Blot for COL10A1
Real validated COL10A1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-COL10A1 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
COL10A1 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 | ~66.2 kDa | |
| Observed band | ~66 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 | Cleaved | |
| Caveat | Homotrimeric assembly | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated COL10A1 Western Blot Protocols
The M01026 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human fetal skin lysate (catalog M01026) |
| 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 | M01026; 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 COL10A1 Western Blot Band Size?
COL10A1 has a predicted full-length mass of 66.2 kDa and an observed band near 66 kDa; the small difference has no established cause.
| Band near 66 kDa | Matches the empirical COL10A1 band; confirm identity with controls |
| Band near 66.2 kDa | Consistent with the predicted full-length chain mass |
| Slightly lower band | Could reflect removal of the 1–18 signal peptide |
| Little or no band in whole-cell lysate | Consistent with secretion into the extracellular matrix |
💡Expected COL10A1 appearanceThe predicted full-length mass is 66.2 kDa, and an approximately 66 kDa band was observed under reducing conditions; confirm band identity with appropriate controls.
| Predicted full-length mass | Sets a 66.2 kDa reference for one chain |
| Signal peptide at residues 1–18 | Contributes to the full-length precursor mass |
| Signal peptide cleavage | Makes the mature chain smaller than the precursor; migration is unverified |
| Homotrimer assembly | Could yield a higher apparent species if intact trimers persist during electrophoresis; this has not been demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | COL10A1 is secreted into the extracellular matrix | Check conditioned medium or extracellular matrix material alongside lysate |
| Band higher than expected | An intact homotrimer is possible, but its migration is unverified | Compare reducing and nonreducing samples and confirm band identity |
| Band lower than expected | Signal peptide cleavage could reduce chain mass | Compare with a suitable full-length control and verify band identity |
| Multiple bands | Precursor and signal-cleaved chains are possible; their separation is unverified | Confirm identities with a second antibody or an appropriate control |
| Weak or no signal | Secretion may leave little COL10A1 in the tested lysate | Test extracellular material and a positive control |
Experiment evidence
Western blot images showing COL10A1 band
M01026 · COL10A1
M01026 · COL10A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for COL10A1 Western blot
🧪For positive controls for COL10A1 in Western blot, you can use a validated positive sample, but the supplied HPA evidence identifies none.
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: COL10A1 is secreted, so conditioned medium may provide a stronger signal than whole-cell lysate.
HPA tissue expression evidence for COL10A1
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 COL10A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for COL10A1, answered from its protein features.
How should COL10A1 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 isoforms explain multiple COL10A1 bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore do not support assigning additional bands to annotated isoforms.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of COL10A1?
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 COL10A1?
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 M01026 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 secreted COL10A1 be quantified across samples?
Quantitation · COL10A1 is annotated as secreted and located in the extracellular space and matrix. Compare the same sample fraction and loading basis across conditions; a change in one fraction may reflect distribution as well as abundance.
How should I interpret a COL10A1 band near 66 kDa?
Interpretation · The supplied predicted mass is 66.2 kDa, and the observed band is approximately 66 kDa. This agreement supports the expected assignment, but band position alone does not confirm identity.
UniProt annotates a signal peptide at residues 1–18 of the 680-residue sequence. Processing could affect the mature chain’s mass, but this feature alone does not establish a visible band shift.
Hydroxylation appears as a keyword, but no modified residue coordinates are supplied. The keyword alone cannot establish the modification state of your sample or explain a visible shift.
UniProt describes COL10A1 as a homotrimer, so an oligomeric species is a possible interpretation. The annotation does not show that a particular higher band is a trimer; verify its identity before assigning it.
UniProt annotates a signal peptide at residues 1–18 of the 680-residue sequence. Processing could affect the mature chain’s mass, but this feature alone does not establish a visible band shift.
Hydroxylation appears as a keyword, but no modified residue coordinates are supplied. The keyword alone cannot establish the modification state of your sample or explain a visible shift.
UniProt describes COL10A1 as a homotrimer, so an oligomeric species is a possible interpretation. The annotation does not show that a particular higher band is a trimer; verify its identity before assigning it.
Boster reagents
COL10A1 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-Collagen X COL10A1 Rabbit Monoclonal Antibody
Cat # M01026
M01026 is a rabbit monoclonal anti-COL10A1 antibody listed for human, mouse, and rat. Its supplied Western blot image shows Collagen X expression in human fetal skin lysate; no mouse or rat WB image is provided.
Which to pick: M01026 is the only listed option. Its documented WB example uses human fetal skin lysate, making that the closest supported sample context. Mouse and rat reactivity is listed, but the supplied image does not demonstrate WB performance in those species.
Source: BosterBio COL10A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q03692.
- Human Protein Atlas. COL10A1 tissue expression.
- PMC5865565 — target-verified WB comparison
- PMC9949800 — target-verified WB comparison
- PMC6113209 — target-verified WB comparison