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- Table of Contents
SLC1A5 / Neutral amino acid transporter B(0) · Western blot design guide
Design a Western Blot for SLC1A5
Real validated SLC1A5 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC1A5 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
SLC1A5 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 | ~56.6 kDa | |
| Observed band | ~55–70 kDa | |
| Gel | 10% (catalog A01474-1) | |
| Positive control | Bronchus (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 Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated SLC1A5 Western Blot Protocols
The A01474-1 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 Jurkat (catalog A01474-1) |
| Gel % | 10% (catalog A01474-1) |
| Load | 30 ug; reducing conditions (catalog A01474-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01474-1) |
| Membrane | nitrocellulose membrane (catalog A01474-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01474-1) |
| Primary antibody | A01474-1 · 0.5 μg/mL (catalog A01474-1) |
| Primary incubation | overnight at 4°C (catalog A01474-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01474-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01474-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01474-1) |
| Detection | ECL (catalog A01474-1) |
Section 2
What Is the Expected SLC1A5 Western Blot Band Size?
SLC1A5 is predicted at 56.6 kDa and observed at ~55–70 kDa; the cause of the apparent range is not established.
| Band at ~55–70 kDa | Empirical SLC1A5 band reported in reducing whole-cell lysates; confirm identity with a specific control |
| Band near 56.6 kDa | Consistent with the predicted protein mass; confirm identity with a specific control |
| Several bands near the expected region | Isoforms 1, 2 and 3 could contribute, but distinct migration is unproven |
| Broader band near the expected region | Variable modification at N-linked sites is possible, but its effect on migration is unproven |
💡Expected SLC1A5 appearanceUniProt predicts 56.6 kDa, while reducing Western blots report ~55–70 kDa; confirm band identity with a specific control because the cause of this apparent range is unestablished.
| UniProt predicted mass | 56.6 kDa is the reference size; the empirical band is ~55–70 kDa |
| N-linked glycosylation at Asn163 | May affect apparent size; the site alone does not establish a visible shift |
| N-linked glycosylation at Asn212 | May affect apparent size; the site alone does not establish a visible shift |
| Isoforms 1, 2 and 3 | Could migrate differently; their size order and resolution are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-localized SLC1A5 may be poorly recovered | Check membrane-protein extraction and a positive lysate control |
| Band higher than expected | N-linked glycosylation may affect migration, but its contribution is unproven | Compare treated and untreated samples and confirm identity with a specific control |
| Band lower than expected | Isoform migration is unknown; band identity is uncertain | Confirm identity with knockdown or another specific control |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible, but unverified | Compare treated and untreated samples and check loading |
| Multiple bands | Isoforms 1, 2 and 3 are annotated, but distinct bands are unproven | Check which bands respond to a specific identity control |
| Weak or no signal | Membrane protein recovery or expression may be low in the sample | Check extraction, loading and a positive lysate control |
Experiment evidence
Western blot images showing SLC1A5 band
A01474-1 · SLC1A5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC1A5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC1A5 in Western blot, you can use bronchus tissue, which HPA rates as high expression.
Positive control: Bronchus (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 the samples.
⚠️Feasibility: As a multi-pass membrane protein, SLC1A5 may require efficient membrane-protein extraction.
HPA tissue expression evidence for SLC1A5
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 |
|---|---|---|---|---|
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | distal tubules | High | Protein (IHC) | HPA → |
| Nasopharynx | basal cells | High | Protein (IHC) | HPA → |
| Placenta | cytotrophoblasts | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC1A5 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC1A5, answered from its protein features.
How should SLC1A5 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 SLC1A5 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 3 lacks residues 1–228, while isoform 2 replaces residues 1–203 with methionine. These differences could affect apparent size and antibody recognition. Check whether the antibody epitope is present in each isoform; the stated coordinates use canonical UniProt numbering.
Which glycosylation sites matter when interpreting SLC1A5 bands?
PTM · UniProt lists N-linked sites at canonical Asn163 and Asn212. Both lie within the region missing from isoform 3; Asn163 also lies within the region replaced in isoform 2. Consider isoform identity when comparing glycosylation patterns. Listed sites do not by themselves prove that glycosylation explains an observed band.
UniProt lists N-acetylmethionine at position 1 and phosphorylation at Ser493, Thr494, Ser503, Ser535, and Ser539, using canonical coordinates. Record treatment conditions when comparing bands, but do not assign a visible shift to these modifications without experimental evidence.
UniProt lists N-acetylmethionine at position 1 and phosphorylation at Ser493, Thr494, Ser503, Ser535, and Ser539, using canonical coordinates. Record treatment conditions when comparing bands, but do not assign a visible shift to these modifications without experimental evidence.
Does this guide establish induction of SLC1A5?
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 SLC1A5 Western blot?
Transfer · SLC1A5 is a multi-pass cell-membrane protein. Verify transfer by checking recovery across the roughly 55–70 kDa observed range, and optimize transfer conditions empirically for your gel and membrane. The supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01474-1 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 SLC1A5 bands be quantified?
Quantitation · Use a consistent band region across samples and document which band or bands were included. Three listed isoforms and two N-linked glycosylation sites can complicate band assignment; an intensity change alone cannot identify the contributing form. Confirm that the quantified signal matches the form your antibody recognizes.
Why might SLC1A5 migrate above its predicted mass?
Interpretation · The predicted mass is 56.6 kDa, while reported bands span about 55–70 kDa. UniProt lists N-linked glycosylation at canonical Asn163 and Asn212, but those features alone do not establish the cause or size of a band shift. Compare treated and untreated samples if testing glycosylation as an explanation.
First compare each band with the roughly 55–70 kDa observed range and the 56.6 kDa predicted mass. Check the antibody epitope against isoforms 1–3, especially the N-terminal changes in isoforms 2 and 3. Glycosylation and the probable homotrimer are additional possibilities to test; neither feature alone identifies an unexpected band.
First compare each band with the roughly 55–70 kDa observed range and the 56.6 kDa predicted mass. Check the antibody epitope against isoforms 1–3, especially the N-terminal changes in isoforms 2 and 3. Glycosylation and the probable homotrimer are additional possibilities to test; neither feature alone identifies an unexpected band.
Boster reagents
SLC1A5 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-ASCT2/SLC1A5 Antibody Picoband®
Cat # A01474-1
The catalog reports one anti-SLC1A5 antibody for Western blot, A01474-1, with reported human and rat reactivity. Its product image shows a band around 55–70 kDa, compared with an expected 57 kDa. The supplied evidence is limited to this product blot.
Which to pick: A01474-1 is the only listed option and has a Western blot image using human HeLa, 293T, Jurkat and HEL cell lysates and rat lung lysate. Use those tested samples and the caption’s conditions to judge its fit for your experiment.
Source: BosterBio SLC1A5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15758.
- Human Protein Atlas. SLC1A5 tissue expression.
- PMC8995533 — target-verified WB comparison
- PMC13430941 — target-verified WB comparison
- PMC11157896 — target-verified WB comparison