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- Table of Contents
SLC39A8 / Metal cation symporter ZIP8 · Western blot design guide
Design a Western Blot for SLC39A8
Source-linked SLC39A8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC39A8 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
SLC39A8 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 | ~49.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Breast (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SLC39A8 Western Blot Protocol Options
The A04007 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human spleen tissue lysate (catalog A04007) |
| 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 | A04007 · (A) 1 and (B) 2 μg/mL (catalog A04007) |
| 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 SLC39A8 Western Blot Band Size?
ZIP8 has a predicted 49.6 kDa precursor; signal cleavage, N-linked glycosylation, isoforms, and homodimer formation could affect migration, but no empirical band is supplied.
| Band near 49.6 kDa | Consistent with the predicted precursor mass; identity needs validation. |
| Band below 49.6 kDa | Could reflect removal of the 1–22 signal peptide; mature migration is unknown. |
| Band above 49.6 kDa | Could reflect N-linked glycosylation at Asn40 or Asn88; a visible shift is unproven. |
| Band near twice the monomer size | Could represent a homodimer preserved during sample preparation. |
| Several bands at different positions | Could include isoforms 1, 2, and 3; their migration differences are unknown. |
💡Expected SLC39A8 appearanceUniProt predicts a 49.6 kDa precursor; no empirical band size is supplied, so validate any candidate band with antibody and sample controls before assigning it to ZIP8.
| Predicted precursor mass | 49.6 kDa is the sequence-based reference, not a validated blot position. |
| N-linked glycosylation at Asn40 | May increase apparent size if modified; the size of any shift is unknown. |
| N-linked glycosylation at Asn88 | May increase apparent size if modified; the size of any shift is unknown. |
| Homodimer formation | May yield a band near twice the monomer size if the complex survives preparation. |
| Isoforms 1, 2, and 3 | May differ in size, but distinct bands are not established. |
| Signal peptide at residues 1–22 | Cleavage would make the mature protein smaller than the precursor; its migration is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ZIP8 is a cell and lysosome membrane protein that may be poorly recovered. | Check membrane protein extraction and include a positive sample. |
| Band higher than expected | Glycosylation or a preserved homodimer may affect migration. | Compare preparation conditions and confirm band identity with an independent antibody. |
| Band lower than expected | Signal peptide cleavage may lower the mature protein mass. | Confirm identity with an independent antibody; do not infer mature size from the precursor alone. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible, but a smear is not established. | Compare with a deglycosylated aliquot and check antibody specificity. |
| Multiple bands | Isoforms or retained homodimers are possible; nonspecific binding also needs exclusion. | Compare sample preparation conditions and validate bands with an independent antibody. |
| Weak or no signal | Recovery of this multi-pass membrane protein may be low. | Check extraction, loading, and a positive control sample. |
Experiment evidence
Western blot images showing SLC39A8 band
A04007 · SLC39A8
A04007-1 · SLC39A8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC39A8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC39A8 in Western blot, you can use breast tissue, which HPA scores as High.
Positive control: Breast (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA lists no Not-detected tissue, so use siRNA knockdown or a KO line for a clean negative control.
HPA tissue expression evidence for SLC39A8
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 |
|---|---|---|---|---|
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Lung | alveolar cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Low | Protein (IHC) | HPA → |
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SLC39A8 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC39A8, answered from its protein features.
How should SLC39A8 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 SLC39A8 isoforms could produce different bands?
Isoforms · UniProt lists three isoforms. Isoform 2 lacks canonical residues 1–67, including the annotated Asn40 glycosylation site, and replaces residues 68–72. Isoform 3 replaces canonical residues 423–460. Check which isoform your antibody can recognize before assigning bands; these sequence differences do not establish where each isoform will migrate.
How can glycosylation affect band interpretation?
PTM · UniProt lists N-linked glycosylation at Asn40 and Asn88, using canonical sequence coordinates. Compare untreated and deglycosylated aliquots if testing whether glycosylation contributes to a band pattern. A change would need experimental confirmation; the site annotations alone do not predict its size.
Does this guide establish induction of SLC39A8?
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 SLC39A8 Western blot?
Transfer · UniProt identifies SLC39A8 as a 49.6 kDa multi-pass membrane protein at the cell and lysosome membranes. The features do not specify a transfer method. Assess transfer efficiency for the bands in your samples and adjust the method empirically.
How should blocking be optimized for SLC39A8?
Blocking · No supplied feature identifies an optimal blocker. Compare blocking conditions using the same membrane preparation and antibody, then choose the condition that gives a clear specific signal with low background. Confirm candidate bands with an appropriate specificity control, especially if several bands appear.
What matters when quantifying SLC39A8 bands?
Quantitation · Keep sample preparation and the measured band consistent across samples. SLC39A8 is annotated at cell and lysosome membranes and has three isoforms plus two canonical N-linked glycosylation sites. State whether quantitation combines bands or measures one band, since the supplied features do not establish that distinct bands represent equal amounts of the same protein.
Should SLC39A8 run at its predicted 49.6 kDa?
Interpretation · Use 49.6 kDa as a sequence-based reference, not an expected band position. UniProt annotates a signal peptide at residues 1–22 and N-linked glycosylation at Asn40 and Asn88. These features alone do not establish a visible shift or explain any observed mass difference; no empirical band position was supplied.
Check antibody recognition of isoforms 1–3, compare untreated and deglycosylated samples, and use a specificity control. Interpret results against the annotated signal peptide (1–22), canonical glycosylation sites (Asn40 and Asn88), and isoform sequence changes. UniProt also annotates a homodimer, but these features alone cannot identify an unexpected band.
Check antibody recognition of isoforms 1–3, compare untreated and deglycosylated samples, and use a specificity control. Interpret results against the annotated signal peptide (1–22), canonical glycosylation sites (Asn40 and Asn88), and isoform sequence changes. UniProt also annotates a homodimer, but these features alone cannot identify an unexpected band.
Boster reagents
SLC39A8 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-ZIP8 SLC39A8 Antibody
Cat # A04007
Real WB data
Anti-ZIP8/SLC39A8 Antibody Picoband®
Cat # A04007-1
Two the supplier anti-SLC39A8 antibodies have WB images: A04007 with human spleen lysate, and A04007-1 with human HEL, rat lung, and mouse lung lysates. A04007-1 shows a band near 80 kDa, although the expected size is 50 kDa.
Which to pick: For human spleen lysate, A04007 has a WB example at 1–2 μg/mL. For the listed human HEL or rat and mouse lung samples, A04007-1 has a WB example at 0.5 μg/mL; assess its band-size discrepancy.
Source: BosterBio SLC39A8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9C0K1.
- Human Protein Atlas. SLC39A8 tissue expression.
- PMC10457451 — target-verified WB comparison
- PMC13508867 — target-verified WB comparison