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- Table of Contents
SLC8A1 / Sodium/calcium exchanger 1 · Western blot design guide
Design a Western Blot for SLC8A1
Source-linked SLC8A1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC8A1 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
SLC8A1 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 | ~108.5 kDa | |
| Observed band | ~140 kDa | |
| Gel | 8% (catalog M03876) | |
| Positive control | Caudate (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 + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked SLC8A1 Western Blot Protocol Options
The M03876 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, rat heart, mouse brain, mouse heart (catalog M03876) |
| Gel % | 8% (catalog M03876) |
| Load | 30 ug; reducing conditions (catalog M03876) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M03876) |
| Membrane | nitrocellulose membrane (catalog M03876) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M03876) |
| Primary antibody | M03876 · 1:500 (catalog M03876) |
| Primary incubation | overnight at 4°C (catalog M03876) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M03876) |
| Secondary incubation | 1.5 hour at RT (catalog M03876) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M03876) |
| Detection | ECL (catalog M03876) |
Section 2
What Is the Expected SLC8A1 Western Blot Band Size?
SLC8A1 has a predicted precursor mass of 108.5 kDa and an empirical band near 140 kDa; the cause of their difference is not established.
| Band near 140 kDa | Empirical SLC8A1-reactive band under reducing conditions; confirm identity with an independent antibody or knockdown. |
| Band below the full-length precursor | Could reflect cleavage of the signal peptide at residues 1–35; its migration is not established. |
| Additional discrete bands | Could reflect isoforms 1, 3, 7, 10, or 5; distinct band positions are not established. |
| Slight shift or diffuse band | N-linked glycosylation at Asn44 and Asn160 could affect migration; a visible effect is not established. |
💡Expected SLC8A1 appearanceUniProt predicts 108.5 kDa for the precursor, while antibody QC reports a band near 140 kDa under reducing conditions; the cause of the difference is unestablished, so confirm identity with an independent antibody or knockdown.
| UniProt predicted precursor mass | 108.5 kDa calculated, versus an empirical band near 140 kDa; the cause of the difference is unestablished. |
| N-linked glycosylation at Asn44 | May alter apparent mobility; the size of any effect is unknown. |
| N-linked glycosylation at Asn160 | May alter apparent mobility; the size of any effect is unknown. |
| Signal peptide at residues 1–35 | Cleavage would make the mature protein smaller than the full-length precursor; its band position is unknown. |
| Isoforms 1, 3, 7, 10, and 5 | Sequence differences may change size, but relative masses and distinct migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SLC8A1 is a multi-pass cell-membrane protein and may be poorly recovered in whole-cell lysate. | Check membrane enrichment and use a known positive lysate. |
| Band higher than expected | The reported band is near 140 kDa despite a predicted precursor mass of 108.5 kDa; its cause is unknown. | Compare with a known positive sample and confirm identity by knockdown or an independent antibody. |
| Band lower than expected | Signal-peptide cleavage or an isoform may contribute, but band positions are not established. | Check antibody epitope coverage and confirm the band with an independent antibody. |
| Broad smear instead of sharp band | Variable N-linked glycosylation at Asn44 or Asn160 is possible but unproven. | Compare treated and untreated samples after deglycosylation and confirm signal specificity. |
| Multiple bands | Five isoforms are listed, but their migration patterns are unknown. | Use an independent antibody or knockdown to identify SLC8A1-reactive bands. |
| Weak or no signal | Recovery of the cell-membrane protein may be low. | Assess membrane extraction and include a known positive sample. |
Experiment evidence
Western blot images showing SLC8A1 band
M03876 · SLC8A1
A03876-2 · SLC8A1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC8A1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC8A1 in Western blot, you can use caudate tissue, where HPA reports high expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a multi-pass membrane protein, SLC8A1 may require membrane-enriched lysate for a strong signal.
HPA tissue expression evidence for SLC8A1
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 → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Placenta | trophoblastic cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC8A1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC8A1, answered from its protein features.
How should SLC8A1 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 SLC8A1 isoforms account for multiple bands?
Isoforms · Five isoforms are listed: 1, 3, 7, 10 and 5. Isoform 3 lacks residues 652–679; isoforms 7 and 5 lack 652–656. Isoforms 3, 7 and 10 also have substitutions at 605–613 and 619–645. Check which sequence the antibody recognizes before assigning a band to an isoform; these sequence differences alone do not establish band positions.
Which glycosylation sites should I consider when interpreting SLC8A1 bands?
PTM · UniProt lists N-linked glycosylation at Asn44 and Asn160. Compare treated and untreated samples under otherwise matched conditions to assess whether glycosylation contributes to a band difference. A shift would need to be observed; the listed sites alone do not predict one.
UniProt annotates phosphoserine at positions 285 and 392. These are UniProt coordinates; check the numbering convention before comparing them with an antibody or paper. The annotations alone do not show that phosphorylation changes the visible band.
UniProt annotates phosphoserine at positions 285 and 392. These are UniProt coordinates; check the numbering convention before comparing them with an antibody or paper. The annotations alone do not show that phosphorylation changes the visible band.
Does this guide establish induction of SLC8A1?
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 SLC8A1?
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 M03876 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 I quantify SLC8A1 when several bands are present?
Quantitation · Define the band or bands being measured consistently across samples and report which species were included. SLC8A1 has five listed isoforms and two N-linked glycosylation sites, so an unassigned band should not automatically be combined with the ~140 kDa band. Confirm band identity before interpreting intensity differences as changes in SLC8A1 abundance.
Why might SLC8A1 appear near 140 kDa instead of 108.5 kDa?
Interpretation · The supplied apparent band is ~140 kDa, while the predicted mass is 108.5 kDa. SLC8A1 is a multi-pass membrane glycoprotein with N-linked sites at Asn44 and Asn160. These features warrant checking glycosylation and band identity, but their presence alone does not establish the cause or size of the difference.
UniProt lists a signal peptide at residues 1–35. Check whether a proposed band assignment assumes processing of that region, but do not infer a specific apparent mass or attribute a smaller band to cleavage from this annotation alone. Use UniProt coordinates when comparing sequence positions.
Compare them with the ~140 kDa observed band and assess antibody recognition of the listed isoforms. The annotated signal peptide, N-linked sites at Asn44 and Asn160, and phosphoserines at 285 and 392 offer possibilities to investigate. None of these features alone identifies an unexpected band or proves a visible shift.
UniProt lists a signal peptide at residues 1–35. Check whether a proposed band assignment assumes processing of that region, but do not infer a specific apparent mass or attribute a smaller band to cleavage from this annotation alone. Use UniProt coordinates when comparing sequence positions.
Compare them with the ~140 kDa observed band and assess antibody recognition of the listed isoforms. The annotated signal peptide, N-linked sites at Asn44 and Asn160, and phosphoserines at 285 and 392 offer possibilities to investigate. None of these features alone identifies an unexpected band or proves a visible shift.
Boster reagents
SLC8A1 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-NCX1 Rabbit Monoclonal Antibody
Cat # M03876
Real WB data
Anti-NCX1/SLC8A1 Antibody Picoband®
Cat # A03876-2
Both listed anti-SLC8A1 antibodies have Western blot images. M03876 shows a ~140 kDa band in rat and mouse brain and heart lysates, versus 109 kDa expected. A03876-2 shows a ~120 kDa band in human K562 lysate, matching its expected size.
Which to pick: For human K562 samples, A03876-2 has the directly relevant image. For mouse or rat brain or heart, choose M03876 based on its demonstrated lanes. M03876 also lists human reactivity, but the supplied image does not test a human sample.
Source: BosterBio SLC8A1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P32418.
- Human Protein Atlas. SLC8A1 tissue expression.
- PMC9283055 — target-verified WB comparison
- PMC10701601 — target-verified WB comparison
- PMC7507321 — target-verified WB comparison