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- Table of Contents
COL11A1 / Collagen alpha-1(XI) chain · Western blot design guide
Design a Western Blot for COL11A1
Real validated COL11A1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-COL11A1 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
COL11A1 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 | ~181.1 kDa | |
| Gel | 8–10% (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 | Processing-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated COL11A1 Western Blot Protocols
The M02909 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | K562 cell lysate (catalog M02909) |
| 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 | M02909; 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 COL11A1 Western Blot Band Size?
COL11A1 has a predicted 181.1 kDa full-length mass; processing, glycosylation, isoforms, and chain association may affect migration, but no empirical band is supplied.
| Band near 181.1 kDa | compatible with the predicted full-length precursor; identity requires confirmation |
| Band below 181.1 kDa | may reflect signal-peptide or propeptide cleavage |
| Little or no band in whole-cell lysate | COL11A1 is secreted into the extracellular matrix |
| Several bands at different positions | may reflect processing or isoforms A, B, C, and 4; their migration is unestablished |
| Higher-mass band under nonreducing conditions | may reflect disulfide-linked chain association |
💡Expected COL11A1 appearanceUniProt predicts a 181.1 kDa full-length precursor, but no empirical band size is supplied; processing and other listed features may affect migration, so confirm band identity with appropriate controls.
| UniProt full-length precursor mass | predicts 181.1 kDa before processing |
| Signal peptide at residues 1–35 | cleavage can lower mature-chain mass |
| Propeptide at residues 36–511 | cleavage can lower mature-chain mass |
| N-linked glycosylation at Asn1640 | may alter apparent migration; a visible shift is unestablished |
| Interchain disulfide at residue 1630 | may retain a higher-mass chain complex under nonreducing conditions |
| Isoforms A, B, C, and 4 | may differ in size, but distinct band positions are unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | COL11A1 is secreted into the extracellular matrix | examine extracellular matrix material and include a positive control |
| Band higher than expected | disulfide-linked chain association may persist | compare reducing and nonreducing samples |
| Band lower than expected | signal-peptide or propeptide cleavage may occur | check whether the antibody recognizes the retained region and compare a positive control |
| Multiple bands | processing or isoforms A, B, C, and 4 may contribute | compare a positive control and confirm bands with an independent antibody |
| Weak or no signal | limited COL11A1 in the sampled material may reduce signal | check extracellular matrix material and verify the antibody with a positive control |
| Fragments below expected size | propeptide cleavage or additional fragmentation may produce smaller species | check antibody epitope location and compare freshly prepared samples |
Experiment evidence
Western blot images showing COL11A1 band
M02909 · COL11A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for COL11A1 Western blot
🧪For positive controls for COL11A1 in Western blot, you can use a confirmed COL11A1-positive sample; the supplied HPA data identifies no specific tissue or cell line.
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: COL11A1 is secreted into the extracellular matrix, so whole-cell lysate may give little signal; consider conditioned medium.
HPA tissue expression evidence for COL11A1
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 COL11A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for COL11A1, answered from its protein features.
Does propeptide removal determine the expected COL11A1 band size?
Band shift · UniProt marks residues 1–35 as signal peptide and 36–511 as propeptide. Check the antibody’s epitope against these regions when interpreting detection of precursor or processed material. The feature coordinates alone do not establish which form appears on the blot or its apparent mass.
How could COL11A1 isoforms affect band interpretation?
Isoforms · UniProt lists isoforms A, B, C, and 4. Relative to the canonical sequence, isoform B replaces residues 261–299, isoform C lacks 261–299, and isoform 4 lacks 300–415. Check whether the antibody epitope is retained in the isoform of interest before assigning bands. These are UniProt coordinates; antibody or publication numbering may differ.
Which COL11A1 modifications matter when evaluating bands?
PTM · UniProt lists allysine at residues 612 and 1452, N-linked glycosylation at Asn1640, and six disulfide bonds. Record these UniProt coordinates when comparing antibody information; other numbering conventions may differ. Their presence alone does not predict a visible shift or identify an unexpected band.
Does this guide establish induction of COL11A1?
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 COL11A1 Western blot?
Transfer · COL11A1 is predicted at 181.1 kDa, so choose and verify a transfer setup that retains detection of proteins around that size. Inspect transfer using a high molecular weight marker. The supplied features do not specify a membrane, transfer chemistry, or expected band position.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02909 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.
Which sample fraction is relevant for COL11A1 quantitation?
Quantitation · COL11A1 is annotated as secreted and located in the extracellular space and matrix. Define whether the measurement targets secreted material, matrix material, or both, and use the same fraction across samples. State that choice when reporting band intensity.
Why might COL11A1 migrate differently from its predicted 181.1 kDa?
Interpretation · The 181.1 kDa prediction describes the full-length sequence. UniProt lists a signal peptide at residues 1–35, a propeptide at 36–511, one N-linked glycosylation site, and other modifications. These features warrant checking which form the antibody detects, but they do not establish an apparent band size or explain a specific shift; no observed band is supplied.
Compare each band with the antibody epitope and the listed isoform changes, signal peptide, propeptide, and disulfide bonds. COL11A1 also occurs in collagen XI trimers with other chains. These features offer possibilities to investigate, but the supplied record has no observed band size and cannot assign an unexpected band.
Compare each band with the antibody epitope and the listed isoform changes, signal peptide, propeptide, and disulfide bonds. COL11A1 also occurs in collagen XI trimers with other chains. These features offer possibilities to investigate, but the supplied record has no observed band size and cannot assign an unexpected band.
Boster reagents
COL11A1 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-COL11A1 Rabbit Monoclonal Antibody
Cat # M02909
M02909 is a rabbit monoclonal anti-COL11A1 antibody listed as reactive with human samples. Its Western blot image shows COL11A1 expression in K562 cell lysate; the supplied evidence does not establish performance in other sample types.
Which to pick: M02909 is the only listed option. It has a Western blot image from K562 cell lysate and is listed for human reactivity; assess suitability for other specimens separately.
Source: BosterBio COL11A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P12107.
- Human Protein Atlas. COL11A1 tissue expression.
- PMC11133424 — target-verified WB comparison
- PMC9167123 — target-verified WB comparison
- PMC4695149 — target-verified WB comparison