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- Table of Contents
COL15A1 / Collagen alpha-1(XV) chain · Western blot design guide
Design a Western Blot for COL15A1
Source-linked COL15A1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-COL15A1 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
COL15A1 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 | ~141.7 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Colon (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked COL15A1 Western Blot Protocol Options
The A05581 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HEK293T (catalog A05581) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A05581; 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 COL15A1 Western Blot Band Size?
COL15A1 has a predicted 141.7 kDa precursor; cleavage, glycosylation and trimerization could affect migration, but no empirical band size is supplied.
| Band near 141.7 kDa | May represent the predicted precursor polypeptide; confirm identity with controls |
| Band above 141.7 kDa | May reflect the listed glycosylation sites; the size effect is unmeasured |
| Very high band under nonreducing conditions | May represent the disulfide-linked trimer |
| Band below the precursor position | May reflect removal of the signal peptide at residues 1–27 |
| Broad band or smear | May reflect variation in chondroitin sulfate modification at the listed serines |
| Little or no band in whole-cell lysate | Consistent with secretion into the extracellular matrix |
💡Expected COL15A1 appearanceUniProt predicts a 141.7 kDa precursor, but no empirical band size is supplied; secretion, signal-peptide cleavage, glycosylation and disulfide-linked trimers complicate migration, so confirm band identity with controls.
| Predicted precursor mass | 141.7 kDa is the sequence-based reference, not a measured band |
| Signal peptide at residues 1–27 | Cleavage may place the mature chain below the precursor |
| Chondroitin sulfate sites at Ser243, Ser247, Ser343 and Ser745 | Modification may raise and vary apparent size |
| O-linked site at Thr265 | Modification may alter apparent size; its effect is unmeasured |
| N-linked sites at Asn306, Asn324 and Asn687 | Modification may raise apparent size; the shift is unmeasured |
| Disulfide-linked trimer | May migrate much higher under nonreducing or incompletely reducing conditions |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | COL15A1 is secreted into the extracellular matrix | Check conditioned medium or extracellular matrix material |
| Band higher than expected | Glycosylation or a disulfide-linked trimer may affect migration | Compare reducing and nonreducing lanes and confirm identity with an independent antibody |
| Band lower than expected | Signal-peptide cleavage may remove residues 1–27 | Check whether the antibody recognizes the mature chain and confirm identity with a second epitope |
| Broad smear instead of sharp band | Chondroitin sulfate modification may vary | Compare appropriately treated and untreated samples with identity controls |
| Multiple bands | Monomer and disulfide-linked trimer may coexist after incomplete reduction | Compare fully reducing and nonreducing conditions |
| Weak or no signal | Secreted COL15A1 may be scarce in the sampled lysate | Enrich extracellular material and include a positive control |
Experiment evidence
Western blot images showing COL15A1 band
A05581 · COL15A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for COL15A1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for COL15A1 in Western blot, you can use colon tissue lysate.
Positive control: Colon (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: Because COL15A1 is secreted into the extracellular matrix, whole-cell lysates may give weak signal.
HPA tissue expression evidence for COL15A1
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 |
|---|---|---|---|---|
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Skin | endothelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced COL15A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for COL15A1, answered from its protein features.
How should COL15A1 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 alternative isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore provide no isoform assignment for additional bands. Compare bands across sample fractions and reducing conditions before assigning an identity.
Which glycosylation sites are relevant to COL15A1 blots?
PTM · UniProt lists chondroitin sulfate O-linked sites at Ser243, Ser247, Ser343 and Ser745; another O-linked site at Thr265; and N-linked sites at Asn306, Asn324, Asn687, Asn807, Asn814 and Asn1046. These are UniProt coordinates; check the numbering convention before comparing them with an antibody or paper.
Does this guide establish induction of COL15A1?
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 COL15A1 Western blot?
Transfer · The supplied features do not specify a transfer method. Because COL15A1 is a 141.7 kDa predicted collagen and can form a disulfide-linked trimer, check transfer efficiency for the mass range you intend to measure, including high-mass material under nonreducing conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05581 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 COL15A1 bands?
Quantitation · Quantify the same defined band or band region across samples, using the same sample fraction and reducing condition. COL15A1 is secreted, glycosylated and trimeric, so document which species your measurement includes; the supplied features do not establish that every band represents an equal amount of protein.
How should I interpret a band above the predicted 141.7 kDa?
Interpretation · 141.7 kDa is the predicted mass of the full sequence, which includes the 1–27 signal peptide. COL15A1 is secreted and has 11 annotated glycosylation sites. These features warrant checking the apparent mass, but they do not establish that a visible shift will occur or explain a specific band.
COL15A1 is annotated as a disulfide-linked trimer and has two disulfide bonds. Compare reducing and nonreducing samples when interpreting high-mass species. These annotations support testing for a condition-dependent pattern; they do not predict exact band positions.
COL15A1 is annotated as secreted and located in the extracellular matrix. Examine the extracellular or matrix-containing fraction relevant to your experiment, alongside the fraction you normally assay. Record the fraction used when comparing band intensity.
Compare their behavior across extracellular and matrix-containing fractions and under reducing and nonreducing conditions. Consider the annotated signal peptide, glycosylation sites and disulfide-linked trimer when interpreting the pattern. None of these features alone identifies an unexpected band or proves its cause.
COL15A1 is annotated as a disulfide-linked trimer and has two disulfide bonds. Compare reducing and nonreducing samples when interpreting high-mass species. These annotations support testing for a condition-dependent pattern; they do not predict exact band positions.
COL15A1 is annotated as secreted and located in the extracellular matrix. Examine the extracellular or matrix-containing fraction relevant to your experiment, alongside the fraction you normally assay. Record the fraction used when comparing band intensity.
Compare their behavior across extracellular and matrix-containing fractions and under reducing and nonreducing conditions. Consider the annotated signal peptide, glycosylation sites and disulfide-linked trimer when interpreting the pattern. None of these features alone identifies an unexpected band or proves its cause.
Boster reagents
COL15A1 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 alpha-1(XV) chain COL15A1 Antibody
Cat # A05581
A05581 is a human-reactive anti-COL15A1 polyclonal antibody with a Western blot image using HEK293T whole cell lysate at 1:500 dilution. The supplied evidence covers this sample and condition only.
Which to pick: A05581 is the only listed option. Its Western blot image uses HEK293T whole cell lysate at 1:500; consider it for human samples while checking suitability for your own sample and protocol.
Source: BosterBio COL15A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P39059.
- Human Protein Atlas. COL15A1 tissue expression.
- PMC10074075 — target-verified WB comparison
- PMC9660072 — target-verified WB comparison
- PMC11294514 — target-verified WB comparison