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- Table of Contents
F10 / Coagulation factor X · Western blot design guide
Design a Western Blot for F10
Real validated F10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-F10 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
F10 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 | Cleaved fragment; mass unreported | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| Lead specificity | Cleaved target; verify fragment size for this antibody | |
| PTM | Glycosylated + Cleaved | |
| Caveat | Activation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated F10 Western Blot Protocols
The A00641 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat cells (catalog A00641) |
| 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 | A00641; 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 F10 Western Blot Band Size?
F10 has a predicted 54.7 kDa precursor; processing and glycosylation could alter migration, but no band position is demonstrated for the lead's activated heavy-chain target.
| Band near 54.7 kDa | Possible F10 precursor; its migration and recognition by the cleavage-specific lead antibody are unverified. |
| Smaller band detected by the lead antibody | Possible activated heavy chain bearing the catalog Cleaved-Ile235 epitope; its mass is unspecified. |
| Band changes between reducing and nonreducing lanes | Processed heavy and light chains are joined by interchain disulfide bonds. |
| Little or no band in cell lysate | F10 is secreted, so cellular abundance may be low. |
| Shifted or diffuse band | Glycosylation is possible at Thr199, Thr211, Asn221 and Asn231; a visible shift or smear is unproven. |
💡Expected F10 appearanceUniProt predicts a 54.7 kDa precursor, but supplies no empirical band; the lead antibody recognizes the activated heavy-chain Cleaved-Ile235 state, whose migration requires band-identity controls.
| 54.7 kDa predicted precursor mass | Provides a sequence-based reference, not a validated Western-blot position. |
| Signal peptide and propeptide cleavage | Removal of residues 1–40 makes the mature protein smaller than the precursor; its apparent mass is unspecified. |
| Two-chain processing and interchain disulfide bonds | Arg excision creates heavy and light chains that remain linked without reduction and can separate upon reduction. |
| O-linked glycosylation at Thr199 and Thr211 | May affect apparent migration; the size of any shift is unestablished. |
| N-linked glycosylation at Asn221 and Asn231 | May affect apparent migration; the size of any shift is unestablished. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted F10 may be scarce in cells. | Check conditioned medium alongside a suitable positive control. |
| Band higher than expected | Linked heavy and light chains or glycosylation may affect migration. | Compare reducing and nonreducing lanes and verify identity with a second antibody. |
| Band lower than expected | The lead antibody targets activated heavy chain rather than intact precursor. | Compare an activation control and an antibody to a different F10 epitope. |
| Broad smear instead of sharp band | Heterogeneous glycosylation is possible but unconfirmed. | Compare matched samples before and after deglycosylation and verify band identity. |
| Multiple bands | F10 processing and glycosylation could produce different species; the lead detects a cleavage-specific state. | Compare reducing conditions, activation controls and a second F10 antibody. |
| Weak or no signal | The Cleaved-Ile235 epitope may be absent if F10 is not activated. | Run a confirmed activated F10 control and check the sample fraction. |
| Fragments below expected size | Activation creates a heavy chain, and further proteolysis is possible. | Compare an activated control and a protected preparation with protease inhibitors. |
Experiment evidence
Western blot images showing F10 band
A00641 · F10
A00641 · F10
M00641-1 · F10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for F10 Western blot
🧪For positive controls for F10 in Western blot, you can use no HPA-supported tissue or cell sample because none is provided.
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: F10 is secreted, so conditioned medium may be more suitable than whole-cell lysate; HPA provides no tissue control candidates.
HPA tissue expression evidence for F10
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 F10 Western Blot Tips
Deeper troubleshooting and optimisation questions for F10, answered from its protein features.
How should F10 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 annotated F10 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Its annotated precursor processing and disulfide-linked chains are relevant when assessing multiple bands; the record does not establish the identity of any observed band.
Which F10 glycosylation sites matter when interpreting bands?
PTM · UniProt lists O-linked glycosylation at Thr199 and Thr211 and N-linked glycosylation at Asn221 and Asn231. Use these UniProt precursor coordinates when comparing site-specific results; paper or antibody numbering may differ. These sites alone do not prove a visible band shift.
UniProt lists 4-carboxyglutamate at positions 46, 47, 54, 56, 59, 60, 65, 66, 69, 72, and 79, plus (3R)-3-hydroxyaspartate at 103. These are UniProt precursor coordinates; check the numbering convention of any antibody or paper. Their presence does not establish a detectable band shift.
UniProt lists 4-carboxyglutamate at positions 46, 47, 54, 56, 59, 60, 65, 66, 69, 72, and 79, plus (3R)-3-hydroxyaspartate at 103. These are UniProt precursor coordinates; check the numbering convention of any antibody or paper. Their presence does not establish a detectable band shift.
Does this guide establish induction of F10?
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 F10?
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 A00641 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 F10 on a Western blot?
Quantitation · Define whether the antibody detects the precursor or a processed chain, then quantify the same band consistently across samples. F10 is secreted and processed into disulfide-linked chains, so record sample type and reducing conditions when comparing results. The supplied features do not identify an observed band for quantitation.
Why might F10 differ from its predicted 54.7 kDa mass?
Interpretation · 54.7 kDa refers to the 488-residue precursor. UniProt lists a signal peptide at 1–31, a propeptide at 32–40, glycosylation, and processing into two chains. These features affect which F10 species is present, but they do not establish an apparent band position or explain a measured difference; no observed band was supplied.
UniProt describes two chains formed from a single-chain precursor by excision of two Arg residues. The chains remain linked by one or more disulfide bonds. Compare reducing and nonreducing lanes when interpreting bands, and check which chain the antibody recognizes before assigning a band to F10.
Check its apparent size against the 54.7 kDa precursor estimate, the signal peptide at 1–31, propeptide at 32–40, and documented two-chain processing. Compare reducing and nonreducing conditions and confirm the antibody's recognized region. UniProt features suggest possibilities but cannot identify an unexpected band by themselves.
UniProt describes two chains formed from a single-chain precursor by excision of two Arg residues. The chains remain linked by one or more disulfide bonds. Compare reducing and nonreducing lanes when interpreting bands, and check which chain the antibody recognizes before assigning a band to F10.
Check its apparent size against the 54.7 kDa precursor estimate, the signal peptide at 1–31, propeptide at 32–40, and documented two-chain processing. Compare reducing and nonreducing conditions and confirm the antibody's recognized region. UniProt features suggest possibilities but cannot identify an unexpected band by themselves.
Boster reagents
F10 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-Cleaved-Factor Xa activated HC (I235) F10 Antibody
Cat # A00641
Real WB data
Anti-Factor X Rabbit Monoclonal Antibody
Cat # M00641-1
Two the supplier anti-F10 antibodies are listed with Western blot images: A00641 in rat cells and M00641-1 in HepG2 cell lysate. These examples document specific tested samples; they do not establish performance across every listed reactive species or sample type.
Which to pick: For rat cell samples, consider A00641, whose WB image uses rat cells and whose listed reactivity includes human, mouse, and rat. For HepG2 lysate, consider M00641-1, which has a WB image in that sample and lists human reactivity.
Source: BosterBio F10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P00742.
- Human Protein Atlas. F10 tissue expression.
- PMC11559047 — target-verified WB comparison