This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
FANCA / Fanconi anemia group A protein · Western blot design guide
Design a Western Blot for FANCA
Real validated FANCA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FANCA 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
FANCA 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 | ~162.8 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated FANCA Western Blot Protocols
The M03662 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A431 cell lysate (catalog M03662) |
| 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 | M03662; 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 FANCA Western Blot Band Size?
FANCA's predicted mass is 162.8 kDa; its isoforms could affect band patterns, but no empirical migration or distinct isoform bands are established.
| Band near 162.8 kDa | Consistent with the predicted FANCA mass; identity needs confirmation |
| Several bands at different positions | Could include isoforms 1, 2, and 3; their migration is not established |
| Band in nuclear fraction | Consistent with FANCA's major nuclear localization |
| Band in cytoplasmic fraction | Consistent with FANCA's minor cytoplasmic localization |
💡Expected FANCA appearanceFANCA has a predicted mass of 162.8 kDa, but no empirical band size is supplied; confirm any candidate band with appropriate identity controls.
| Predicted FANCA mass | 162.8 kDa provides a reference size, not a measured migration |
| Isoform 1 | Relative size and migration are not supplied |
| Isoform 2 | May differ in size from other isoforms; relative size is unknown |
| Isoform 3 | May differ in size from other isoforms; relative size is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FANCA is predominantly nuclear and may be poorly recovered | Check extraction and compare a nuclear fraction with loading controls |
| Band higher than expected | Identity or migration of the band is unverified | Confirm with an independent antibody or FANCA-deficient control |
| Band lower than expected | An alternative isoform is possible, but its size is unknown | Compare with a FANCA-deficient control and check isoform coverage |
| Multiple bands | Isoforms 1, 2, and 3 are listed, but distinct bands are unproven | Test band identity with a FANCA-deficient control |
| Weak or no signal | The major FANCA form is nuclear | Check nuclear extraction and sample loading |
| Fragments below expected size | Fragment identities are not established by the supplied features | Check sample integrity and confirm bands with a FANCA-deficient control |
Experiment evidence
Western blot images showing FANCA band
M03662 · FANCA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FANCA Western blot
🧪For positive controls for FANCA in Western blot, you can use a FANCA-expressing sample, but the supplied HPA evidence identifies no specific positive tissue or cell line.
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 sample.
⚠️Feasibility: Without HPA expression data, suitable tissue controls cannot be confirmed.
HPA tissue expression evidence for FANCA
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 FANCA Western Blot Tips
Deeper troubleshooting and optimisation questions for FANCA, answered from its protein features.
How should FANCA 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 FANCA isoform 2 produce a smaller band?
Isoforms · UniProt lists residues 298–1455 as missing in isoform 2, leaving a much shorter sequence than canonical FANCA. A smaller band could therefore be consistent with isoform 2 if the antibody recognizes its retained region; the features do not establish where it migrates.
Isoform 3 replaces canonical residues 1390–1455 with a different sequence. An antibody recognizing that canonical C-terminal region may detect isoforms differently. Check the antibody epitope against the listed sequences before comparing bands.
Isoform 3 replaces canonical residues 1390–1455 with a different sequence. An antibody recognizing that canonical C-terminal region may detect isoforms differently. Check the antibody epitope against the listed sequences before comparing bands.
Does phosphorylation explain a shifted FANCA band?
PTM · UniProt lists phosphoserine at position 1449 in the canonical sequence. This site lies in the region missing from isoform 2 and replaced in isoform 3. Feature presence alone does not show a visible shift or explain an observed-versus-calculated mass difference. Keep UniProt numbering explicit when comparing antibody or paper coordinates.
Does this guide establish induction of FANCA?
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 FANCA Western blot?
Transfer · FANCA's predicted canonical mass is 162.8 kDa. Check transfer of proteins near that size with a molecular-weight marker and membrane staining, and adjust transfer conditions if recovery is poor. The supplied features do not specify one required method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03662 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 FANCA 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.
Where should full-length FANCA appear on a Western blot?
Interpretation · The canonical sequence predicts 162.8 kDa. No observed band position is supplied, so use this as a reference rather than an exact apparent mass. The listed features do not establish a visible shift.
UniProt describes the major FANCA form as nuclear and a minor form as cytoplasmic. Keep fraction identity and loading consistent when quantifying signals, and interpret a weaker cytoplasmic band in that context. The location annotation does not provide an expected nuclear-to-cytoplasmic ratio.
Consider the listed isoforms: isoform 2 lacks residues 298–1455, while isoform 3 has a replacement at residues 1390–1455. Check whether the antibody epitope is retained in each. The listed phosphoserine at canonical position 1449 does not, by itself, establish a separate visible band.
UniProt describes the major FANCA form as nuclear and a minor form as cytoplasmic. Keep fraction identity and loading consistent when quantifying signals, and interpret a weaker cytoplasmic band in that context. The location annotation does not provide an expected nuclear-to-cytoplasmic ratio.
Consider the listed isoforms: isoform 2 lacks residues 298–1455, while isoform 3 has a replacement at residues 1390–1455. Check whether the antibody epitope is retained in each. The listed phosphoserine at canonical position 1449 does not, by itself, establish a separate visible band.
Boster reagents
FANCA 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-FANCA Rabbit Monoclonal Antibody
Cat # M03662
The catalog reports one anti-FANCA antibody: M03662, a rabbit monoclonal with reported human reactivity. Its Western blot image shows FANCA expression in A431 cell lysate; the supplied evidence does not establish performance in other samples.
Which to pick: M03662 is the only listed option. Choose it for a human FANCA Western blot when the A431 cell lysate example is relevant; check suitability for other sample types separately.
Source: BosterBio FANCA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O15360.
- Human Protein Atlas. FANCA tissue expression.
- PMC11460194 — target-verified WB comparison
- PMC9369391 — target-verified WB comparison
- PMC6476309 — target-verified WB comparison