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- Table of Contents
FTH1 / Ferritin heavy chain · Western blot design guide
Design a Western Blot for FTH1
Real validated FTH1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FTH1 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
FTH1 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 | ~21.2 kDa | |
| Observed band | ~19 kDa | |
| Gel | 5–20% (catalog M02401) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Parathyroid gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated FTH1 Western Blot Protocols
The M02401 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human 293T, human MCF-7 (catalog M02401) |
| Gel % | 5–20% (catalog M02401) |
| Load | 30 ug; reducing conditions (catalog M02401) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02401) |
| Membrane | nitrocellulose membrane (catalog M02401) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02401) |
| Primary antibody | M02401 · 1:500 (catalog M02401) |
| Primary incubation | overnight at 4°C (catalog M02401) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M02401) |
| Secondary incubation | 1.5 hour at RT (catalog M02401) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02401) |
| Detection | ECL (catalog M02401) |
Section 2
What Is the Expected FTH1 Western Blot Band Size?
FTH1 is predicted at 21.2 kDa and observed near 19 kDa in reducing blots; the cause of the difference is not established.
| Band near 19 kDa | Empirical FTH1 band observed in reducing whole-cell and tissue lysates |
| Band near 21.2 kDa | Near the UniProt predicted mass; confirm identity with an antibody control |
| Closely spaced bands near the main band | Could reflect the listed N-terminal processing or phosphorylation, but distinct migration is unproven |
| Additional lower band | May warrant checking N-terminal processing or degradation; no fragment size is established |
| High-mass band under poorly dissociating conditions | Could reflect persistence of the listed 24-subunit ferritin complex |
💡Expected FTH1 appearanceUniProt predicts 21.2 kDa, while antibody QC shows an approximately 19 kDa band in reducing lysates; the difference is unexplained, so confirm band identity with appropriate controls.
| UniProt predicted mass | 21.2 kDa for FTH1; antibody QC observes approximately 19 kDa |
| N-terminal processing at Thr2 | Removes the initiating residue; no measurable migration effect is established |
| N-terminal acetylation at Met1 or processed Thr2 | Is documented, but no visible size shift is established |
| Phosphoserine 179 | May alter migration, but no distinct band is demonstrated |
| Phosphoserine 183 | May alter migration, but no distinct band is demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | FTH1 is listed in cytoplasm, lysosome, and autophagosomes; absence from whole-cell lysate has no established feature-specific explanation | Check lysate quality and an FTH1-positive control |
| Band higher than expected | The listed 24-subunit complex may persist if dissociation is incomplete | Check denaturation conditions and compare with a reducing control |
| Band lower than expected | The observed band is approximately 19 kDa versus a 21.2 kDa prediction; its offset is unexplained | Compare with the QC band and confirm identity using an independent antibody or FTH1 depletion |
| Multiple bands | N-terminal processing and phosphorylation are listed, but distinct bands are unproven | Confirm each band with an independent antibody or FTH1 depletion |
| Weak or no signal | FTH1 detection may depend on sample abundance and fraction selection | Check loading, antibody performance, and a positive lysate |
| Fragments below expected size | No defined FTH1 fragment mass is supplied | Check sample integrity and use an independent antibody to assess identity |
Experiment evidence
Western blot images showing FTH1 band
M02401 · FTH1
M02401-2 · FTH1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FTH1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FTH1 in Western blot, you can use bone marrow tissue, which shows high HPA expression.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Parathyroid gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Parathyroid gland is not detected by HPA, but confirm its negative status by Western blot.
HPA tissue expression evidence for FTH1
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 → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced FTH1 Western Blot Tips
Deeper troubleshooting and optimisation questions for FTH1, answered from its protein features.
How should FTH1 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 an FTH1 isoform explain an additional band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for an additional band.
Which phosphorylation sites should I consider?
PTM · UniProt lists phosphoserine at positions 179 and 183. Keep those UniProt coordinates when comparing site-specific results; other numbering conventions may differ. Their presence does not establish a visible band shift.
Does this guide establish induction of FTH1?
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 FTH1?
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 M02401 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.
Does sample fraction matter when quantifying FTH1?
Quantitation · FTH1 is listed in the cytoplasm, lysosome, and autophagosome. Compare the same sample fraction and the same defined band across samples, and report whether quantitation refers to whole-cell material or a compartment-enriched fraction.
Why does FTH1 appear near 19 kDa instead of 21.2 kDa?
Interpretation · The supplied blot observation is approximately 19 kDa, while the predicted mass is 21.2 kDa. Treat 19 kDa as an apparent migration position; the listed features do not establish the cause of the difference.
UniProt lists N-acetylmethionine at position 1 and N-acetylthreonine at position 2 in N-terminally processed ferritin heavy chain. These are UniProt coordinates. The features alone do not establish a visible shift or explain the 19-kDa band.
Ferritin is described as a 24-subunit oligomer containing heavy and light chains. Consider that assembly context when evaluating higher-mass bands, but do not assign a band to an oligomer from migration alone.
UniProt lists N-acetylmethionine at position 1 and N-acetylthreonine at position 2 in N-terminally processed ferritin heavy chain. These are UniProt coordinates. The features alone do not establish a visible shift or explain the 19-kDa band.
Ferritin is described as a 24-subunit oligomer containing heavy and light chains. Consider that assembly context when evaluating higher-mass bands, but do not assign a band to an oligomer from migration alone.
Boster reagents
FTH1 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-Ferritin FTH1 Rabbit Monoclonal Antibody
Cat # M02401
Real WB data
Anti-Ferritin Heavy Chain Rabbit Monoclonal Antibody
Cat # M02401-2
Two the supplier rabbit monoclonal antibodies are listed for FTH1 Western blotting. M02401 has a blot image showing a band near 19 kDa in the named human, rat, and mouse samples; M02401-2 has a HeLa lysate blot image. No publication evidence is supplied.
Which to pick: Choose M02401 for its reported human, mouse, and rat reactivity and blot image across named samples from all three species. M02401-2 reports human reactivity and shows a HeLa lysate blot; consider it for that context.
Source: BosterBio FTH1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P02794.
- Human Protein Atlas. FTH1 tissue expression.
- PMC12811428 — target-verified WB comparison
- PMC6038692 — target-verified WB comparison
- PMC11715977 — target-verified WB comparison