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- Table of Contents
SLC39A14 / Metal cation symporter ZIP14 · Western blot design guide
Design a Western Blot for SLC39A14
Source-linked SLC39A14 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC39A14 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
SLC39A14 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~54.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Glycosylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SLC39A14 Western Blot Protocol Options
The A04761 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A04761; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | 1 h (catalog A04761) |
| 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 SLC39A14 Western Blot Band Size?
ZIP14 has a predicted 54.2 kDa precursor; glycosylation, signal peptide cleavage, and isoforms could affect migration, but no band position is demonstrated.
| Band near 54.2 kDa | Consistent with the predicted ZIP14 precursor size; identity requires controls. |
| Band above 54.2 kDa | N-linked glycosylation at Asn77, Asn87, or Asn102 could affect migration; no shift is established. |
| Band below 54.2 kDa | Signal peptide cleavage could yield a smaller mature protein; its band size is unknown. |
| Several bands at different positions | Isoforms 1, 2, and 3 are documented, but distinct bands are not established. |
💡Expected SLC39A14 appearanceZIP14 has a predicted 54.2 kDa precursor mass, while N-linked glycosylation, signal peptide cleavage, and isoforms may affect migration; no empirical band size is supplied, so confirm band identity with controls.
| Predicted precursor mass | 54.2 kDa is the sequence-based reference, not a measured band. |
| N-linked glycosylation at Asn77 | Could alter apparent size; the shift is unknown. |
| N-linked glycosylation at Asn87 | Could alter apparent size; the shift is unknown. |
| N-linked glycosylation at Asn102 | Could alter apparent size; the shift is unknown. |
| Signal peptide at residues 1–30 | Cleavage could make mature ZIP14 smaller than its precursor. |
| Isoforms 1, 2, and 3 | Their sizes may differ, but relative migration is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ZIP14 may be poorly recovered or solubilized. | Check membrane enrichment and extraction, then verify antibody performance with a positive control. |
| Band higher than expected | N-linked glycosylation could affect migration. | Compare matched samples before and after N-glycan removal and verify band identity. |
| Band lower than expected | Signal peptide cleavage could produce a smaller mature protein. | Check whether the antibody recognizes the detected region and confirm identity with a positive control. |
| Broad smear instead of sharp band | Different N-linked glycoforms could contribute, though a smear is not established. | Compare matched samples before and after N-glycan removal. |
| Multiple bands | Isoforms 1, 2, and 3 are documented, but their migration is unknown. | Check transcript expression and confirm candidate bands with an independent antibody or ZIP14 depletion. |
| Weak or no signal | Recovery of this multi-pass membrane protein may be low. | Check membrane extraction and include a positive-control lysate. |
Experiment evidence
Western blot images showing SLC39A14 band
A04761 · SLC39A14
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC39A14 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC39A14 in Western blot, you can use appendix tissue, which HPA rates High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside.
⚠️Feasibility: As a multi-pass membrane protein, SLC39A14 may give a clearer signal in a membrane-enriched sample.
HPA tissue expression evidence for SLC39A14
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Kidney | distal tubules | High | Protein (IHC) | HPA → |
| Rectum | glandular cells | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC39A14 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC39A14, answered from its protein features.
How should SLC39A14 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 ZIP14 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 2, and 3. Relative to the canonical sequence, isoform 2 replaces residues 156–199, while isoform 3 replaces residues 445–492. Check whether the antibody recognizes a region shared by the isoforms before assigning separate bands to them. These UniProt coordinates use canonical numbering.
How can glycosylation be assessed on a ZIP14 blot?
PTM · Compare untreated and N-glycan–removed samples using the same antibody. UniProt lists N-linked sites at Asn77, Asn87, and Asn102 in canonical-sequence numbering. A mobility change would support a glycosylation contribution, but the listed sites alone do not establish a visible shift.
Does this guide establish induction of SLC39A14?
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 SLC39A14 Western blot?
Transfer · ZIP14 is annotated as a multi-pass cell-membrane protein with a predicted mass of 54.2 kDa. Start with a transfer setup suited to membrane proteins and check transfer efficiency on the membrane before interpreting a weak or absent band. The supplied features do not identify one required transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04761 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 ZIP14 bands be quantified?
Quantitation · Compare the same assigned band across samples and normalize to a suitable loading measure. ZIP14 is a multi-pass membrane protein with three listed isoforms, so keep sample preparation and the band chosen for quantification consistent. Do not combine distinct bands without evidence that they represent the same form.
Why might ZIP14 migrate differently from its predicted mass?
Interpretation · The 54.2 kDa prediction is for the canonical sequence. ZIP14 has a signal peptide at UniProt residues 1–30 and N-linked glycosylation sites at Asn77, Asn87, and Asn102. Processing or glycosylation could affect migration, but these features alone cannot predict a band position; no observed band is supplied.
UniProt annotates residues 1–30 as a signal peptide in the 492-residue canonical precursor. Account for possible signal-peptide removal when comparing a band with the 54.2 kDa predicted mass. The annotation does not establish the apparent mass of the detected form.
UniProt reports ZIP14 as a homotrimer, but that annotation alone does not identify a high-mass blot band. Compare reducing and sample-preparation conditions, and verify antibody specificity before attributing the band to ZIP14. Glycosylation at UniProt Asn77, Asn87, and Asn102 also cannot, by itself, explain a particular observed mass.
UniProt annotates residues 1–30 as a signal peptide in the 492-residue canonical precursor. Account for possible signal-peptide removal when comparing a band with the 54.2 kDa predicted mass. The annotation does not establish the apparent mass of the detected form.
UniProt reports ZIP14 as a homotrimer, but that annotation alone does not identify a high-mass blot band. Compare reducing and sample-preparation conditions, and verify antibody specificity before attributing the band to ZIP14. Glycosylation at UniProt Asn77, Asn87, and Asn102 also cannot, by itself, explain a particular observed mass.
Boster reagents
SLC39A14 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-ZIP14 SLC39A14 Antibody
Cat # A04761
The catalog reports A04761 for SLC39A14 Western blotting, with stated human, mouse and rat reactivity. Its WB image caption reports tissue lysates from those species, loaded at 10 μg and probed at 2 μg/mL. No publication evidence is supplied.
Which to pick: A04761 is the only listed antibody and has a WB validation image. Its caption reports human, mouse and rat tissue lysates; use those tested contexts and the reported conditions to judge fit for your samples.
Source: BosterBio SLC39A14 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15043.
- Human Protein Atlas. SLC39A14 tissue expression.
- PMC13489016 — target-verified WB comparison
- PMC11532203 — target-verified WB comparison
- PMC11943061 — target-verified WB comparison