This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
COL1A2 / Collagen alpha-2(I) chain · Western blot design guide
Design a Western Blot for COL1A2
Real validated COL1A2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-COL1A2 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
COL1A2 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 | ~129.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Cervix (IHC candidate; verify WB) +4 more | |
| 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 | 1 isoform(s) |
Section 1
Real Curated COL1A2 Western Blot Protocols
The A00624-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | various cells (catalog A00624-1) |
| 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 | A00624-1 · 1:500 (catalog A00624-1) |
| 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 COL1A2 Western Blot Band Size?
COL1A2 has a predicted full-length mass of 129.3 kDa; cleavage and glycosylation could affect migration, but no empirical band position is supplied.
| Band near 129.3 kDa | Consistent with the predicted full-length precursor mass; confirm identity with a control |
| Band below 129.3 kDa | Could reflect signal-peptide or propeptide cleavage |
| Little or no band in whole-cell lysate | COL1A2 is secreted into the extracellular matrix |
| Band above 129.3 kDa | Glycosylation could affect migration, but no shift is established |
| Two bands at different positions | Could reflect precursor and processed forms; band identities need confirmation |
💡Expected COL1A2 appearanceUniProt predicts a 129.3 kDa full-length precursor; processing and glycosylation may affect migration, but no empirical band size is supplied, so confirm candidate bands with identity controls.
| Predicted full-length mass | 129.3 kDa is the sequence-based precursor reference |
| Signal peptide at residues 1–22 | Cleavage could yield a smaller product than the precursor |
| Propeptide at residues 23–79 | Cleavage could further reduce product size |
| Alternate O-linked galactosylation at hydroxylysine 177 | Could affect migration; a visible shift is not established |
| N-linked glycosylation at asparagine 1267 | Could affect migration; the size of any shift is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | COL1A2 is secreted into extracellular matrix | Examine extracellular or matrix material alongside a positive control |
| Band higher than expected | Glycosylation may affect migration, but its contribution is unconfirmed | Test glycosylation effects and verify band identity with an independent antibody |
| Band lower than expected | Signal-peptide or propeptide cleavage may reduce size | Compare precursor and extracellular samples and verify band identity |
| Multiple bands | Precursor and processed forms may coexist | Compare cellular and extracellular samples using a validated positive control |
| Weak or no signal | Secreted protein may be scarce in the sampled fraction | Enrich extracellular or matrix material and include a positive control |
Experiment evidence
Western blot images showing COL1A2 band
A00624-1 · COL1A2
A00624 · COL1A2
A00624 · COL1A2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for COL1A2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for COL1A2 in Western blot, you can use cervix tissue, which HPA rates as highly expressed.
Positive control: Cervix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: COL1A2 is secreted into the extracellular matrix, so whole-cell lysates may give weak signal; use an extracellular matrix sample.
HPA tissue expression evidence for COL1A2
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 |
|---|---|---|---|---|
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Low | Protein (IHC) | HPA → |
Section 3
Advanced COL1A2 Western Blot Tips
Deeper troubleshooting and optimisation questions for COL1A2, answered from its protein features.
How should COL1A2 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.
Do the supplied features support multiple COL1A2 isoforms?
Isoforms · Only one isoform is listed, with no alternative sequence. Do not assign multiple bands to splice isoforms on the basis of these features.
Which annotated COL1A2 residue modifications should I consider?
PTM · UniProt positions include pyrrolidone carboxylic acid at 23 and 80; 4-hydroxyproline at 47, 50, 62, 65, 68, 71, 102, 108, 420, 441, and 444; allysine at 84; and alternate 5-hydroxylysine at 177. These annotations do not establish a visible band shift. Use UniProt numbering when comparing them with antibody or paper coordinates.
UniProt lists alternate O-linked glycosylation of hydroxylysine at position 177 and N-linked glycosylation of asparagine at 1267. Position 177 is also annotated as alternate 5-hydroxylysine. Neither site alone predicts how far a band will migrate.
UniProt lists alternate O-linked glycosylation of hydroxylysine at position 177 and N-linked glycosylation of asparagine at 1267. Position 177 is also annotated as alternate 5-hydroxylysine. Neither site alone predicts how far a band will migrate.
Does this guide establish induction of COL1A2?
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 COL1A2 Western blot?
Transfer · The supplied features give a predicted mass of 129.3 kDa and probable assembly with two alpha-1(I) chains, but specify no transfer method. Check whether the molecular species you intend to measure transfers and is retained under your chosen conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00624-1 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 COL1A2 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might COL1A2 migrate away from its predicted 129.3 kDa?
Interpretation · 129.3 kDa is the predicted sequence mass. COL1A2 has a signal peptide (1..22), a propeptide (23..79), and annotated modifications. These features may affect which molecular species is detected, but they do not establish a visible shift or a specific apparent mass. No observed band size was supplied.
The UniProt features identify a signal peptide at 1..22 and a propeptide at 23..79. Consider whether an antibody recognizes a region retained in the species being measured. These annotations alone cannot identify an unexpected band.
COL1A2 is annotated as secreted and located in the extracellular space and matrix. Compare the same sample fraction and preparation across conditions; a cellular fraction alone may not reflect extracellular COL1A2. The supplied features give no basis for an induction claim.
Consider signal peptide and propeptide processing, annotated modifications, and the probable trimer containing one alpha-2(I) and two alpha-1(I) chains. Three disulfide bonds are listed. These features suggest possibilities to investigate, but cannot establish the identity of any extra band without experimental validation.
The UniProt features identify a signal peptide at 1..22 and a propeptide at 23..79. Consider whether an antibody recognizes a region retained in the species being measured. These annotations alone cannot identify an unexpected band.
COL1A2 is annotated as secreted and located in the extracellular space and matrix. Compare the same sample fraction and preparation across conditions; a cellular fraction alone may not reflect extracellular COL1A2. The supplied features give no basis for an induction claim.
Consider signal peptide and propeptide processing, annotated modifications, and the probable trimer containing one alpha-2(I) and two alpha-1(I) chains. Three disulfide bonds are listed. These features suggest possibilities to investigate, but cannot establish the identity of any extra band without experimental validation.
Boster reagents
COL1A2 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-2(I) chain COL1A2 Antibody
Cat # A00624-1
Real WB data
Anti-Cleaved-COL1A2 (G1102) Antibody
Cat # A00624
Two the supplier anti-COL1A2 antibodies list Human, Mouse, and Rat reactivity and have WB images. Their captions report various cells at 1:500 and VEC cells at 1:1000; these examples do not establish performance across all specimens of the listed species.
Which to pick: Both have WB images and the same listed reactivity. A00624-1 shows various cells at 1:500; A00624 shows VEC cells at 1:1000. Choose based on which reported sample and conditions better match your experiment.
Source: BosterBio COL1A2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P08123.
- Human Protein Atlas. COL1A2 tissue expression.
- PMC7037297 — target-verified WB comparison
- PMC8888114 — target-verified WB comparison
- PMC6096367 — target-verified WB comparison