This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
VEGFC / Vascular endothelial growth factor C · Western blot design guide
Design a Western Blot for VEGFC
Real validated VEGFC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VEGFC 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
VEGFC 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 | ~46.9 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 | Processing-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated VEGFC Western Blot Protocols
The A00623 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | L929 cells (catalog A00623) |
| 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 | A00623; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A00623) |
| 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 VEGFC Western Blot Band Size?
VEGFC has a predicted 46.9 kDa precursor; processing, glycosylation, and dimerization could affect migration, but no empirical band size establishes which band appears.
| Band near 46.9 kDa | Candidate full-length precursor; identity requires confirmation |
| Band below 46.9 kDa | May reflect signal-peptide or propeptide cleavage |
| Band near twice a corresponding monomer band under non-reducing conditions | May reflect an inter-chain disulfide-linked homodimer |
| Higher band or broad smear | May reflect variable N-linked glycosylation at Asn175, Asn205, or Asn240 |
| Little or no band in whole-cell lysate | VEGFC is secreted |
💡Expected VEGFC appearanceThe predicted 46.9 kDa mass describes the precursor; processing and N-linked glycosylation may alter migration, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted precursor mass | The 419-residue sequence has a calculated mass of 46.9 kDa |
| N-linked glycosylation at Asn175, Asn205, and Asn240 | May increase apparent mass or broaden migration; the size effect is unquantified |
| Inter-chain disulfide-linked homodimer | May appear near twice the corresponding monomer mass if disulfide bonds persist |
| Signal peptide at residues 1–31 and propeptide at residues 32–111 | Cleavage can yield smaller products than the precursor; product masses are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted VEGFC may be depleted from cells | Check conditioned medium alongside lysate |
| Band higher than expected | N-linked glycosylation or incomplete reduction of an inter-chain disulfide-linked dimer | Compare reducing conditions and a glycosidase-treated sample |
| Band lower than expected | Signal-peptide or propeptide cleavage may produce a smaller form | Compare precursor and processed-form recognition with a second epitope-specific antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Compare untreated and glycosidase-treated samples |
| Multiple bands | Precursor and processed forms or different glycosylation states may coexist | Compare reducing and glycosidase-treated lanes and confirm band identity with a second antibody |
Experiment evidence
Western blot images showing VEGFC band
A00623 · VEGFC
A00623 · VEGFC
M00623 · VEGFC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VEGFC Western blot
🧪For positive controls for VEGFC in Western blot, you can use a validated VEGFC-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 such as Ponceau alongside the samples.
⚠️Feasibility: VEGFC is secreted, so conditioned medium may provide a better signal than whole-cell lysate.
HPA tissue expression evidence for VEGFC
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 VEGFC Western Blot Tips
Deeper troubleshooting and optimisation questions for VEGFC, answered from its protein features.
How should VEGFC 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 VEGFC bands?
Isoforms · The supplied record lists one isoform and no alternative sequence, so it provides no annotated isoform explanation for multiple bands. Consider processing and glycosylation as possibilities, but verify band identity experimentally.
Which VEGFC glycosylation sites matter when interpreting bands?
PTM · UniProt annotates N-linked glycosylation at Asn175, Asn205, and Asn240. These positions use the supplied full-length sequence numbering; antibody or paper numbering may differ. Glycosylation could affect mobility, but the annotations alone do not establish a visible shift.
Does this guide establish induction of VEGFC?
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 VEGFC?
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 A00623 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 VEGFC secretion affect quantitation?
Quantitation · Because VEGFC is secreted, a cell-lysate measurement alone may not capture the protein released into medium. For comparisons, use a consistent collection period and sample volume, and state whether the measured signal came from medium or lysate.
Why might VEGFC migrate differently from its predicted 46.9 kDa?
Interpretation · 46.9 kDa is the predicted mass for the 419-residue sequence. VEGFC has a signal peptide at residues 1..31, a propeptide at 32..111, and three annotated N-linked glycosylation sites. These features can affect the species detected and its apparent mobility, but no observed band size is supplied, so they cannot establish the cause of a particular difference.
VEGFC has a signal peptide at residues 1..31 and a propeptide at 32..111; its keywords also include cleavage on a pair of basic residues. Consider whether the antibody detects a precursor or a processed species when interpreting a band. The supplied features do not specify observed fragment sizes.
UniProt identifies VEGFC as secreted. Check conditioned medium when assessing expression, and interpret a weak cell-lysate signal in light of secretion. Compare samples collected and handled consistently.
VEGFC is annotated as a noncovalent, antiparallel homodimer with five disulfide bonds. Sample preparation may affect which molecular forms are detected, so record reducing and denaturing conditions when comparing bands. These features alone do not identify any unexpected band.
VEGFC has a signal peptide at residues 1..31 and a propeptide at 32..111; its keywords also include cleavage on a pair of basic residues. Consider whether the antibody detects a precursor or a processed species when interpreting a band. The supplied features do not specify observed fragment sizes.
UniProt identifies VEGFC as secreted. Check conditioned medium when assessing expression, and interpret a weak cell-lysate signal in light of secretion. Compare samples collected and handled consistently.
VEGFC is annotated as a noncovalent, antiparallel homodimer with five disulfide bonds. Sample preparation may affect which molecular forms are detected, so record reducing and denaturing conditions when comparing bands. These features alone do not identify any unexpected band.
Boster reagents
VEGFC 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-VEGF-C Antibody
Cat # A00623
Real WB data
Anti-VEGFC Antibody
Cat # M00623
Two the supplier anti-VEGFC antibodies have Western blot images: A00623 in L929 cells at 1:2000, and M00623 in 293 and CEM cells plus mouse and rat heart lysates at 1:1000. These examples document tested samples, not universal validation.
Which to pick: Both list Human, Mouse, and Rat reactivity. Choose M00623 if its documented cell or heart lysates match your samples; choose A00623 for the documented L929 cell example. The captions do not establish performance in other specimens.
Source: BosterBio VEGFC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P49767.
- Human Protein Atlas. VEGFC tissue expression.
- PMC5997793 — target-verified WB comparison
- PMC13399718 — target-verified WB comparison
- PMC9998469 — target-verified WB comparison