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- Table of Contents
FANCI / Fanconi anemia group I protein · Western blot design guide
Design a Western Blot for FANCI
Real validated FANCI Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FANCI 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
FANCI 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 | ~149.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated FANCI Western Blot Protocols
The A05108 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A05108) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05108) |
| Primary antibody | A05108 · 1:1000 (catalog A05108) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05108) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05108) |
Section 2
What Is the Expected FANCI Western Blot Band Size?
FANCI has a predicted monomer mass of 149.3 kDa; isoforms, phosphorylation, and homodimer formation could affect bands, but their migration is not demonstrated.
| Band near 149.3 kDa | consistent with the predicted FANCI monomer; confirm identity with a specific control |
| Band near 299 kDa | could reflect a retained FANCI homodimer; its migration is not established |
| Several bands at different positions | could reflect FANCI isoforms 1, 2, 3, and 4; distinct migration is not established |
| Doublet near 149.3 kDa | could reflect different phosphorylation states; a visible shift is not established |
💡Expected FANCI appearanceUniProt predicts a 149.3 kDa FANCI monomer, but no empirical band size is supplied; use a FANCI-specific identity control to assess any band near that position.
| Predicted FANCI monomer mass | sets a sequence-based reference of 149.3 kDa |
| FANCI homodimer | could appear near twice the monomer mass if the complex survives sample preparation |
| Isoforms 1 and 2 | may differ in size; individual masses and migration are not supplied |
| Isoforms 3 and 4 | may differ in size; individual masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FANCI may be missed if nuclear protein extraction is inadequate | check nuclear extraction and a FANCI-positive control |
| Band higher than expected | a FANCI homodimer may have survived sample preparation | compare strongly denatured samples and confirm the band with FANCI depletion |
| Band lower than expected | an isoform or FANCI fragment may be detected | confirm with FANCI depletion and an antibody to another region |
| Multiple bands | FANCI isoforms or different phosphorylation states are possible | use FANCI depletion to identify specific bands |
| Weak or no signal | FANCI recovery or antibody detection may be insufficient | check sample loading, nuclear extraction, and a positive control |
| Fragments below expected size | FANCI may have degraded during sample preparation | prepare fresh lysate with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing FANCI band
A05108 · FANCI
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FANCI Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FANCI in Western blot, you can use bone marrow lysate, which HPA rates High.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because FANCI is nuclear, ensure the lysate includes nuclear proteins.
HPA tissue expression evidence for FANCI
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | High | Protein (IHC) | HPA → |
| Skin | melanocytes | High | Protein (IHC) | HPA → |
| Testis | cells in seminiferous ducts | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced FANCI Western Blot Tips
Deeper troubleshooting and optimisation questions for FANCI, answered from its protein features.
How should FANCI 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.
Which FANCI isoforms could produce different bands?
Isoforms · Four isoforms are listed. Relative to the supplied canonical sequence, isoforms 1 and 2 lack residues 819–878; isoform 2 also lacks residue 1117. Isoform 4 lacks residues 253–1328. These differences could affect band size, although the supplied features do not establish which isoforms appear in a sample.
Which phosphorylation sites matter when interpreting FANCI bands?
PTM · UniProt lists phosphoserine at positions 407, 556, 730 and 1121, and phosphothreonine at 952. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish a resolvable band shift.
Does this guide establish induction of FANCI?
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 FANCI Western blot?
Transfer · FANCI’s predicted mass is 149.3 kDa, so choose conditions that transfer a protein of this size and verify transfer in the relevant region. The supplied features do not specify a membrane or transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05108 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 multiple FANCI bands be quantified?
Quantitation · Identify the band being measured and use the same band across samples. The listed alternative sequences mean that distinct isoforms could differ in size, while phosphorylation alone does not establish a separate visible band. The supplied features do not assign an observed band to any isoform.
Should FANCI run at its predicted 149.3 kDa?
Interpretation · 149.3 kDa is the predicted mass; no observed band position is supplied. FANCI has alternative sequences and documented phosphorylation sites, but those features alone do not establish a visible shift or explain any measured difference.
Check whether the antibody epitope overlaps residues 819–878, absent from isoforms 1 and 2, or residues 253–1328, absent from isoform 4. An epitope at residue 1117 is also absent from isoform 2. Detection depends on the antibody’s actual epitope.
Compare their positions with the predicted 149.3 kDa protein and the listed isoform deletions. A smaller band could warrant an isoform check, particularly for isoform 4’s large deletion. Band position alone cannot establish identity, and no observed FANCI band size is supplied.
Check whether the antibody epitope overlaps residues 819–878, absent from isoforms 1 and 2, or residues 253–1328, absent from isoform 4. An epitope at residue 1117 is also absent from isoform 2. Detection depends on the antibody’s actual epitope.
Compare their positions with the predicted 149.3 kDa protein and the listed isoform deletions. A smaller band could warrant an isoform check, particularly for isoform 4’s large deletion. Band position alone cannot establish identity, and no observed FANCI band size is supplied.
Boster reagents
FANCI 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-Fanconi anemia group I protein FANCI Antibody
Cat # A05108
A05108 is listed as human-reactive and has a Western blot image from extracts of various cell lines. The reported conditions include 25 µg of protein per lane and a 1:1000 antibody dilution. The caption does not identify the cell lines.
Which to pick: A05108 is the only listed anti-FANCI antibody. It is listed for human reactivity and has a Western blot image; check whether its reported conditions suit your samples.
Source: BosterBio FANCI gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NVI1.
- Human Protein Atlas. FANCI tissue expression.
- PMC6970268 — target-verified WB comparison
- PMC10703153 — target-verified WB comparison
- PMC5538132 — target-verified WB comparison