This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SLC3A2 / Amino acid transporter heavy chain SLC3A2 · Western blot design guide
Design a Western Blot for SLC3A2
Real validated SLC3A2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SLC3A2 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
SLC3A2 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 | ~57.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebral cortex (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 | 4 isoform(s) |
Section 1
Real Curated SLC3A2 Western Blot Protocols
The A01794-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HEB cells (catalog A01794-1) |
| 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 | A01794-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:20000 (catalog A01794-1) |
| 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 SLC3A2 Western Blot Band Size?
SLC3A2 has a predicted mass of 57.9 kDa; glycosylation, isoforms and heterodimer formation may affect migration, but no empirical band is supplied.
| Band near 57.9 kDa | Consistent with the predicted SLC3A2 mass; confirm its identity with controls. |
| Band above 57.9 kDa | N-linked glycosylation could affect migration, but its visible effect is unestablished. |
| Higher band under non-reducing conditions | Could reflect the disulfide-linked SLC3A2–light-subunit heterodimer. |
| Several bands | Could reflect isoforms or glycosylation states; distinct isoform bands are unestablished. |
💡Expected SLC3A2 appearanceUniProt predicts 57.9 kDa for SLC3A2; glycosylation and disulfide-linked heterodimer formation may alter migration, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted molecular mass | 57.9 kDa is the sequence-based reference, not a measured band. |
| N-linked glycosylation at Asn264 and Asn280 | May affect apparent mass; the size of any shift is unknown. |
| Complex N-linked glycosylation at Asn323 | May affect apparent mass; the size of any shift is unknown. |
| N-linked glycosylation at Asn405 | May affect apparent mass; the size of any shift is unknown. |
| Disulfide-linked heterodimer | May produce a higher band under non-reducing conditions; its mass depends on the light subunit. |
| Isoforms 1, 2, 3 and 5 | May differ in size, but their masses and band separation are unspecified. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-localized SLC3A2 may be poorly extracted. | Check membrane extraction and include a positive control. |
| Band higher than expected | N-linked glycosylation or an incompletely reduced heterodimer may contribute. | Compare deglycosylated and fully reduced samples. |
| Band lower than expected | Reduced glycosylation could affect migration; the band's identity is uncertain. | Compare deglycosylated samples and use an independent SLC3A2 antibody. |
| Broad smear instead of sharp band | Heterogeneous N-linked glycans are possible, but a smear is not established by site annotations. | Compare before and after deglycosylation. |
| Multiple bands | Isoforms or different glycosylation states are possible; their separation is unestablished. | Compare deglycosylated samples and verify bands with an independent antibody. |
| Weak or no signal | Membrane localization may limit recovery in the tested fraction. | Check membrane enrichment, extraction and a positive control. |
Experiment evidence
Western blot images showing SLC3A2 band
A01794-1 · SLC3A2
A01794-4 · SLC3A2
A01794 · SLC3A2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SLC3A2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SLC3A2 in Western blot, you can use cerebral cortex lysate, which shows high HPA expression.
Positive control: Cerebral cortex (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 cell-membrane protein, SLC3A2 may give a stronger signal in membrane-enriched lysates.
HPA tissue expression evidence for SLC3A2
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 |
|---|---|---|---|---|
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Kidney | proximal tubules (cell body) | High | Protein (IHC) | HPA → |
| Pancreas | pancreatic endocrine cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | trophoblastic 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC3A2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SLC3A2, answered from its protein features.
How should SLC3A2 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 SLC3A2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 2, 3 and 5. Its alternative-sequence records describe different N-terminal sequences for isoforms 1, 3 and 5. Check whether the antibody recognizes the sequence present in each isoform before assigning bands; the records do not establish their apparent masses.
Which listed modifications should guide band interpretation?
PTM · UniProt lists N-acetylserine and phosphoserine at position 2; phosphothreonine at 5; phosphoserine at 33, 64, 305, 307, 309 and 426; and the supplied annotation “Phosphoserine N-acetylmethionine” at 430. These are UniProt coordinates, which may differ from antibody or paper numbering. Their presence does not demonstrate a visible band shift.
Does this guide establish induction of SLC3A2?
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 SLC3A2?
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 A01794-1 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.
What should be controlled when quantifying SLC3A2?
Quantitation · Use the same sample preparation and reducing conditions across samples, and quantify the same resolved band or defined set of bands consistently. SLC3A2 is membrane-associated, has four listed isoforms and forms a disulfide-linked heterodimer; changes in recovery or band selection could complicate comparisons.
Why might SLC3A2 migrate differently from its predicted mass?
Interpretation · The predicted mass is 57.9 kDa. UniProt lists N-linked glycosylation at Asn264, Asn280, Asn323 and Asn405, which may affect apparent migration. No observed band position is supplied, and these features alone do not establish a visible shift.
SLC3A2 forms a disulfide-linked heterodimer with a catalytic light subunit, and UniProt lists one disulfide bond. Compare reducing and nonreducing samples when assessing a higher-mass band: disruption of the intersubunit linkage could change its migration. These features alone cannot identify a particular band.
Consider the listed disulfide-linked heterodimer with a non-glycosylated light subunit, especially if the band changes under reducing conditions. The listed N-linked sites may also affect SLC3A2 migration. Neither feature proves the identity of an unexpected band.
UniProt places SLC3A2 at the cell membrane, including the apical cell membrane, and lists a transmembrane helix and signal-anchor keyword. Include membrane-containing material when preparing samples, and keep the fraction used consistent across comparisons.
SLC3A2 forms a disulfide-linked heterodimer with a catalytic light subunit, and UniProt lists one disulfide bond. Compare reducing and nonreducing samples when assessing a higher-mass band: disruption of the intersubunit linkage could change its migration. These features alone cannot identify a particular band.
Consider the listed disulfide-linked heterodimer with a non-glycosylated light subunit, especially if the band changes under reducing conditions. The listed N-linked sites may also affect SLC3A2 migration. Neither feature proves the identity of an unexpected band.
UniProt places SLC3A2 at the cell membrane, including the apical cell membrane, and lists a transmembrane helix and signal-anchor keyword. Include membrane-containing material when preparing samples, and keep the fraction used consistent across comparisons.
Boster reagents
SLC3A2 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-CD98 SLC3A2 Antibody
Cat # A01794-1
Real WB data
Anti-CD98/SLC3A2 Antibody
Cat # A01794-4
Real WB data
Anti-SLC3A2 Antibody
Cat # A01794
Three the supplier anti-SLC3A2 antibodies have Western blot images. A01794-4 shows human SiHa, A549, MCF-7 and HeLa and rat C6 lysates, with a reported 75–120 kDa band versus an expected 68/71 kDa. These examples do not establish universal validation.
Which to pick: For human or rat lysates, A01794-4 has the most detailed WB example, including sample lanes and conditions. A01794 shows human HEB lysate. A01794-1 lists human, mouse and rat reactivity, but its image caption identifies only HEB cells.
Source: BosterBio SLC3A2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P08195.
- Human Protein Atlas. SLC3A2 tissue expression.
- PMC3749780 — target-verified WB comparison
- PMC13317265 — target-verified WB comparison
- PMC5687629 — target-verified WB comparison