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- Table of Contents
PROC / Vitamin K-dependent protein C · Western blot design guide
Design a Western Blot for PROC
Real validated PROC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PROC 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
PROC 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 | ~52.1 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 + Phosphorylated | |
| Caveat | Glycosylation and cleavage | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PROC Western Blot Protocols
The M01742 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M01742) |
| 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 | M01742; 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 PROC Western Blot Band Size?
PROC has a 52.1 kDa precursor backbone; glycosylation can shift migration upward, while precursor and heavy/light-chain cleavage change the band pattern.
| Band near 52.1 kDa | full-length precursor backbone near its predicted mass |
| Higher or diffuse band | glycosylated protein C, with variable occupancy including partial glycosylation at Asn371 |
| Lower band than the precursor | processed protein C after precursor or chain cleavage |
| Several bands at different positions | processing, glycoforms, or isoforms 1 and 2 may contribute |
| Little or no band in whole-cell lysate | protein C is secreted |
💡Expected PROC appearanceExpect secreted protein C to migrate according to glycosylation and processing relative to its 52.1 kDa precursor backbone; cleavage into disulfide-linked heavy and light chains can change the pattern with reduction.
| 52.1 kDa predicted precursor mass | provides the unmodified full-length reference size |
| N-glycosylation at Asn139, Asn290, and Asn355 | adds carbohydrate and can raise apparent size |
| Partial N-glycosylation at Asn371 | can create variable migration or a broader band |
| O-glycosylation at Thr19 | can modify the propeptide before its removal |
| Signal peptide 1–18 and propeptide 19–42 cleavage | makes the mature protein smaller than the full-length precursor |
| Disulfide-linked heavy and light chains | remain linked without reduction but separate upon reduction |
| Isoforms 1 and 2 | may differ in size; individual masses are unavailable |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | protein C is secreted | check conditioned medium or a secretion-enriched sample |
| Band higher than expected | N-linked glycans increase apparent size | compare with a deglycosylated aliquot |
| Band lower than expected | signal and propeptide removal or chain processing | compare reducing and nonreducing lanes |
| Broad smear instead of sharp band | heterogeneous glycosylation, including partial occupancy at Asn371 | compare with a deglycosylated aliquot |
| Multiple bands | processing, glycoforms, or isoforms 1 and 2 | compare reduction and deglycosylation conditions |
| Fragments below expected size | cleavage into heavy and light chains | compare reducing and nonreducing lanes |
Experiment evidence
Western blot images showing PROC band
M01742 · PROC
PA1682 · PROC
A01742 · PROC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PROC Western blot
🧪For positive controls for PROC in Western blot, you can use a positive sample once identified; the supplied HPA evidence lists 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: PROC is secreted, so whole-cell lysate may give little signal; consider conditioned medium.
HPA tissue expression evidence for PROC
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 PROC Western Blot Tips
Deeper troubleshooting and optimisation questions for PROC, answered from its protein features.
How should PROC 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.
Can bands identify PROC isoforms?
Isoforms · Use the linked UniProt isoform annotations; migration alone does not identify an isoform. No additional isoform-specific band assignment is established here.
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 PROC?
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 PROC Western blot?
Transfer · Plan transfer around the PROC forms you aim to detect: the predicted 52.1 kDa precursor or its processed light and heavy chains. Confirm that transfer retains the relevant bands, since their apparent sizes and an observed band size are not supplied.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01742 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 PROC bands be quantified?
Quantitation · Define the molecular form before quantifying: precursor, processed chains, or thrombin-activated PROC may produce different signals. Use the same sample type and reducing condition across comparisons, and quantify the band recognized by your antibody consistently.
Why might PROC bands differ from the predicted 52.1 kDa?
Interpretation · 52.1 kDa describes the predicted precursor. PROC loses its signal peptide and propeptide, undergoes glycosylation, and is cleaved into light and heavy chains. Interpret band position in light of which form your antibody detects; no observed band size is supplied.
UniProt lists two PROC isoforms, 1 and 2. Check whether the antibody recognizes both before assigning an extra band to an isoform; their expected sizes are not supplied.
PROC is synthesized as one chain and then cleaved into light and heavy chains linked by a disulfide bond. Reducing conditions can separate those chains. Compare reducing and nonreducing lanes, and check the antibody's recognized region when interpreting bands.
UniProt states that thrombin activates PROC by cleavage. If comparing samples with different activation states, interpret new or shifted bands alongside this processing step. The supplied features do not give an expected size for an activated band.
PROC is listed as secreted and also localized to the endoplasmic reticulum and Golgi apparatus. Choose secreted material or cell samples according to whether you want extracellular PROC or intracellular processing forms, and label those sample types separately when comparing bands.
UniProt lists two PROC isoforms, 1 and 2. Check whether the antibody recognizes both before assigning an extra band to an isoform; their expected sizes are not supplied.
PROC is synthesized as one chain and then cleaved into light and heavy chains linked by a disulfide bond. Reducing conditions can separate those chains. Compare reducing and nonreducing lanes, and check the antibody's recognized region when interpreting bands.
UniProt states that thrombin activates PROC by cleavage. If comparing samples with different activation states, interpret new or shifted bands alongside this processing step. The supplied features do not give an expected size for an activated band.
PROC is listed as secreted and also localized to the endoplasmic reticulum and Golgi apparatus. Choose secreted material or cell samples according to whether you want extracellular PROC or intracellular processing forms, and label those sample types separately when comparing bands.
Boster reagents
PROC 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-Protein C Rabbit Monoclonal Antibody
Cat # M01742
Real WB data
Anti-Protein C/PROC Antibody Picoband®
Cat # PA1682
Real WB data
Anti-Protein C/PROC Antibody Picoband®
Cat # A01742
Three the supplier anti-Protein C antibodies are listed for Western blot: M01742, PA1682, and A01742. Each has a WB image. The supplied details identify HepG2 cell lysate for M01742; JURKAT, CEM, SMMC, and HELA cell lysates for PA1682; and rat and mouse liver tissue lysates for A01742.
Which to pick: Match the WB example to your sample context: M01742 for HepG2 cells, PA1682 for the listed cell lysates, or A01742 for rat or mouse liver tissue.
Source: BosterBio PROC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P04070.
- Human Protein Atlas. PROC tissue expression.
- PMC12898814 — target-verified WB comparison
- PMC4115806 — target-verified WB comparison