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- Table of Contents
FUNDC1 / FUN14 domain-containing protein 1 · Western blot design guide
Design a Western Blot for FUNDC1
Real validated FUNDC1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FUNDC1 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
FUNDC1 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 | ~17.2 kDa | |
| Observed band | ~17 kDa | |
| Gel | 12% (catalog A08688-2) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (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 | 1 isoform(s) |
Section 1
Real Curated FUNDC1 Western Blot Protocols
The A08688-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SiHa, human 293T, rat brain (catalog A08688-2) |
| Gel % | 12% (catalog A08688-2) |
| Load | 30 ug; reducing conditions (catalog A08688-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A08688-2) |
| Membrane | nitrocellulose membrane (catalog A08688-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A08688-2) |
| Primary antibody | A08688-2 · 0.5 μg/mL (catalog A08688-2) |
| Primary incubation | overnight at 4°C (catalog A08688-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A08688-2) |
| Secondary incubation | 1.5 hour at RT (catalog A08688-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A08688-2) |
| Detection | ECL (catalog A08688-2) |
Section 2
What Is the Expected FUNDC1 Western Blot Band Size?
FUNDC1 is predicted at 17.2 kDa and observed at approximately 17 kDa; the supplied evidence does not establish a cause for the small difference.
| Band near 17 kDa | Matches the observed FUNDC1 band and its 17.2 kDa predicted mass |
| Faint band near 17 kDa in whole-cell lysate | Mitochondrial outer membrane FUNDC1 may be poorly recovered from the sample |
| Little band near 17 kDa in a soluble fraction | FUNDC1 is a multi-pass mitochondrial outer membrane protein |
| Nearby doublet around 17 kDa | Could reflect phosphorylation states at Ser13, Ser17, or Tyr18, but distinct migration is unproven |
💡Expected FUNDC1 appearanceFUNDC1 has a predicted mass of 17.2 kDa and an observed band at approximately 17 kDa; confirm identity with antibody specificity controls and appropriate mitochondrial membrane recovery.
| UniProt predicted mass | Places full-length FUNDC1 near 17.2 kDa; the observed band is approximately 17 kDa |
| Ser13 phosphorylation by CK2 | May affect migration, but a visible size shift is not established |
| Ser17 phosphorylation by ULK1 | May affect migration, but a visible size shift is not established |
| Tyr18 phosphorylation by SRC | May affect migration, but a visible size shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of multi-pass mitochondrial outer membrane FUNDC1 | Check membrane solubilization and a mitochondrial membrane marker |
| Band higher than expected | Incomplete solubilization or altered migration of the membrane protein is possible | Optimize solubilization and compare with the approximately 17 kDa control band |
| Band lower than expected | A smaller signal has no assigned FUNDC1 form in the supplied features | Check sample integrity and antibody specificity |
| Broad smear instead of sharp band | Incomplete solubilization of the multi-pass membrane protein is possible | Optimize extraction and sample preparation |
| Multiple bands | Phosphorylation states are possible, but distinct bands are unproven | Use phosphatase treatment and antibody specificity controls |
| Weak or no signal | Low recovery of mitochondrial outer membrane FUNDC1 | Check membrane enrichment and loading with a mitochondrial membrane marker |
Experiment evidence
Western blot images showing FUNDC1 band
A08688-2 · FUNDC1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FUNDC1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FUNDC1 in Western blot, you can use caudate tissue, where HPA reports high expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: As a mitochondrial outer-membrane protein, FUNDC1 requires lysate containing mitochondria for a useful signal.
HPA tissue expression evidence for FUNDC1
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FUNDC1 Western Blot Tips
Deeper troubleshooting and optimisation questions for FUNDC1, answered from its protein features.
How should FUNDC1 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 another isoform explain an unexpected FUNDC1 band?
Isoforms · UniProt lists one isoform and no alternative sequence. Do not assign an extra band to a FUNDC1 isoform from these features alone; assess antibody specificity before interpreting it.
Which phosphorylation sites matter when comparing FUNDC1 bands?
PTM · The supplied UniProt coordinates list Ser13 phosphorylated by CK2, Ser17 by ULK1, and Tyr18 by SRC. State that these are UniProt positions if comparing antibody or paper numbering. These modifications may matter to phospho-specific detection, but their presence alone does not establish a visible shift.
Does this guide establish induction of FUNDC1?
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 FUNDC1 Western blot?
Transfer · FUNDC1 is a 17.2 kDa multi-pass mitochondrial outer-membrane protein. Optimize transfer for retention of a small membrane protein, and check both the transferred membrane and post-transfer gel. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A08688-2 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 FUNDC1 signal be quantified?
Quantitation · Compare bands within the expected approximately 17 kDa region using equal loading and consistent membrane-protein extraction. Because FUNDC1 resides in the mitochondrial outer membrane, account for differences in mitochondrial content when comparing samples.
Should FUNDC1 run above its predicted mass?
Interpretation · FUNDC1 is predicted at 17.2 kDa, and the reported band is approximately 17 kDa. Its listed features do not establish a visible band shift. Check a molecular weight marker and compare the band with the expected range.
FUNDC1 interacts with MAP1LC3A, MAP1LC3B, GABARAP, DNM1L/DPR1, and GPX4, but those interactions do not identify a higher band. The supplied features list no glycosylation sites or alternative isoform. Verify band specificity before assigning an identity.
FUNDC1 interacts with MAP1LC3A, MAP1LC3B, GABARAP, DNM1L/DPR1, and GPX4, but those interactions do not identify a higher band. The supplied features list no glycosylation sites or alternative isoform. Verify band specificity before assigning an identity.
Boster reagents
FUNDC1 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-FUNDC1 Antibody Picoband®
Cat # A08688-2
The catalog reports one anti-FUNDC1 antibody for Western blotting, A08688-2, with stated human, mouse and rat reactivity. Its WB image shows an approximately 17 kDa band in the human cell, rat brain and cell, and mouse brain and cell lysates named in the caption. No independent validation is supplied.
Which to pick: A08688-2 is the only listed option. Its WB image includes human SiHa and 293T, rat brain and C6, and mouse brain and RAW264.7 lysates, tested at 0.5 μg/mL primary antibody under the stated conditions. Match your sample and protocol to that evidence.
Source: BosterBio FUNDC1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8IVP5.
- Human Protein Atlas. FUNDC1 tissue expression.
- PMC10504546 — target-verified WB comparison
- PMC13603441 — target-verified WB comparison
- PMC6465113 — target-verified WB comparison