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- Table of Contents
SLC7A5 / Large neutral amino acids transporter small subunit 1 · Western blot design guide
Design a Western Blot for SLC7A5
Real validated SLC7A5 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC7A5 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
SLC7A5 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 | ~55 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | Phosphorylated + Ubl conjugation | |
| Caveat | Reducing-condition controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SLC7A5 Western Blot Protocols
The A01016 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A01016) |
| Primary antibody | A01016; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A01016) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A01016) |
Section 2
What Is the Expected SLC7A5 Western Blot Band Size?
SLC7A5 has a predicted 55 kDa monomer; its disulfide-linked heterodimer may migrate higher, but an empirical band size and migration effect are not supplied.
| Band near 55 kDa | consistent with the predicted SLC7A5 monomer; confirm identity with controls |
| Higher band under nonreducing conditions | may represent the disulfide-linked SLC7A5–SLC3A2 heterodimer |
| Higher band decreases after reduction | consistent with disruption of the disulfide-linked heterodimer |
| Weak band in a soluble fraction | consistent with limited recovery of membrane-localized SLC7A5 |
💡Expected SLC7A5 appearanceUniProt predicts a 55 kDa SLC7A5 monomer, but no empirical band size is supplied; confirm a candidate band with reduction and band-identity controls.
| UniProt predicted monomer mass | sets a 55 kDa reference for SLC7A5 |
| Cys164 inter-chain disulfide | can retain SLC7A5 in a higher-mass complex under nonreducing conditions |
| SLC3A2-linked heterodimer | can migrate above the SLC7A5 monomer; its band size is not supplied |
| Reduction of the disulfide-linked heterodimer | can release the SLC7A5 monomer near its predicted size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | poor recovery of membrane-localized SLC7A5 | check membrane extraction and test a membrane-enriched fraction |
| Band higher than expected | disulfide-linked SLC7A5–SLC3A2 may persist | compare reducing and nonreducing lanes |
| Band lower than expected | band identity or degradation is uncertain | check antibody specificity and compare fresh, protease-inhibited lysate |
| Multiple bands | monomer and disulfide-linked heterodimer may coexist | compare fully reducing and nonreducing lanes and confirm band identity |
| Weak or no signal | limited extraction of the multi-pass membrane protein | check solubilization and load a membrane-enriched fraction |
Experiment evidence
Western blot images showing SLC7A5 band
A01016 · SLC7A5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC7A5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC7A5 in Western blot, you can use adrenal gland tissue, where HPA reports high expression.
Positive control: Adrenal gland (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 multipass membrane protein, SLC7A5 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for SLC7A5
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced SLC7A5 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC7A5, answered from its protein features.
How should SLC7A5 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.
Should I expect multiple SLC7A5 isoform bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to splice isoforms based on this evidence alone.
Which SLC7A5 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at positions 31 and 35 and phosphothreonine at position 45. These are UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a visible shift, so compare matched samples before attributing a band difference to phosphorylation.
Does this guide establish induction of SLC7A5?
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 SLC7A5 Western blot?
Transfer · SLC7A5 is a 507-residue multi-pass membrane protein. Check transfer efficiency by examining target left in the gel and signal on the membrane, then adjust the transfer conditions if needed. The supplied features do not identify one optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01016 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 SLC7A5 across samples?
Quantitation · SLC7A5 localizes to the cell membrane, including the apical membrane. Keep sample fraction and reducing conditions consistent across lanes, and quantify the same identified band within the assay’s linear range. Changes in membrane recovery could otherwise affect the comparison.
Why might the SLC7A5 band differ from 55 kDa?
Interpretation · 55 kDa is the predicted mass; no observed band position is supplied. SLC7A5 is a multi-pass membrane protein with three listed phosphorylation sites and a disulfide-linked partner, but these features alone cannot establish the cause of a mass difference. Compare the band under consistent sample conditions.
SLC7A5 forms a disulfide-linked heterodimer with SLC3A2/4F2hc. Compare reducing and nonreducing samples if a higher band appears; a condition-dependent change would help assess whether the disulfide-linked complex contributes. The supplied features do not specify its gel position.
Check whether bands vary with reducing conditions, given the disulfide-linked SLC3A2/4F2hc heterodimer. The record lists one isoform and no glycosylation sites, so it does not support assigning extra bands to alternative isoforms or listed glycosylation. The phosphorylation sites alone do not prove a visible shift.
SLC7A5 forms a disulfide-linked heterodimer with SLC3A2/4F2hc. Compare reducing and nonreducing samples if a higher band appears; a condition-dependent change would help assess whether the disulfide-linked complex contributes. The supplied features do not specify its gel position.
Check whether bands vary with reducing conditions, given the disulfide-linked SLC3A2/4F2hc heterodimer. The record lists one isoform and no glycosylation sites, so it does not support assigning extra bands to alternative isoforms or listed glycosylation. The phosphorylation sites alone do not prove a visible shift.
Boster reagents
SLC7A5 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-SLC7A5 Antibody
Cat # A01016
The catalog reports one anti-SLC7A5 antibody, A01016, with reported human, mouse, and rat reactivity. Its Western blot image uses HT-1080 cell extracts at a 1:1000 primary dilution; the supplied evidence shows this sample context only.
Which to pick: A01016 is the only listed option and has a Western blot image from HT-1080 extracts. Use the reported 1:1000 primary dilution as a starting point; the supplied image does not establish performance in other samples.
Source: BosterBio SLC7A5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q01650.
- Human Protein Atlas. SLC7A5 tissue expression.
- PMC5788887 — target-verified WB comparison
- PMC11371717 — target-verified WB comparison
- PMC3935884 — target-verified WB comparison