This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SLC1A4 / Neutral amino acid transporter A · IHC design guide
Design Immunohistochemistry for SLC1A4
Use pancreatic exocrine glands as a high-staining reference and adipocytes as a not-detected comparison (HPA tissue IHC). This paraffin IHC guide covers fixation, expected staining, and the catalog antibody’s 0.5–1 μg/ml starting dilution (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC1A4 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic, often membranous in tissue (HPA tissue IHC) | |
| Staining pattern | Various cell types: cytoplasmic, often membranous (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Tissue-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; extracellular vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended SLC1A4 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published SLC1A4 IHC methods for PSP brain sections (PMC6208732) and HCC tissue sections (PMC7991385).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PA2186); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SLC1A4, 0.5-1μg/ml (datasheet PA2186) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC1A4-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, often with a membranous pattern. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval). The PSP study used citrate pH 6 for 30 min (PMC6208732).
Section 2
What Is the Expected SLC1A4 Staining Pattern?
SLC1A4 is an 8-pass cell membrane transporter (UniProt P43007 topology); HPA tissue IHC describes general cytoplasmic staining, often with a membranous pattern (HPA: tissue IHC). Expect signal in selected cells, including pancreatic exocrine glandular cells and cerebral cortex neuropil (HPA: High). HPA rates tissue IHC reliability Enhanced, while reporting medium consistency between staining and RNA expression (HPA: tissue IHC).
| Distinct cytoplasmic staining, sometimes outlining cells, in pancreatic exocrine glands; strong signal in cortex neuropil. | This fits HPA's tissue IHC profile and its High calls for those structures (HPA: tissue IHC). A membrane component also fits the transporter topology (UniProt P43007 topology). Judge the stained cell or structure, rather than treating every cell in a positive tissue as positive. |
| Predominantly nuclear staining, with little cytoplasmic or membrane signal, across the section. | A nuclear-only pattern conflicts with the reported tissue IHC profile and membrane location (HPA: tissue IHC; UniProt P43007 subcellular location). Treat it as suspect staining and compare with a known-positive structure and a control that omits the primary antibody before scoring it. |
| Strong staining in adipocytes or liver cholangiocytes while the expected positive structures stain weakly. | Those specific cell populations are Not detected in HPA tissue IHC (HPA: adipocytes; HPA: cholangiocytes). Check antibody-dependent cross-reactivity and endogenous chromogenic detection activity with suitable controls (general IHC practice). The HPA calls do not make every cell in either tissue negative. |
| A broad, even deposit covers cells and spaces, obscuring cell boundaries. | That pattern cannot establish SLC1A4 localisation or identify the stained cell type. Review the primary-omission control, blocking, antibody concentration, washes and chromogen development (general IHC practice); compare any remaining structured signal with HPA's cytoplasmic and often membranous profile (HPA: tissue IHC). |
| No signal in pancreatic exocrine glandular cells or cerebral cortex neuropil. | These are High HPA tissue IHC examples, so an entirely blank known-positive control weakens the interpretation of negative study sections (HPA: pancreas; HPA: cerebral cortex). Check that the expected structure is present, then review the validated IHC workflow and detection controls (general IHC practice). |
💡Expected SLC1A4 appearanceCall a section positive when the appropriate cells or structures show clear cytoplasmic staining, often with a membrane pattern, and stronger signal in HPA High examples such as pancreatic exocrine cells; widespread nuclear-only or uniform background staining is suspect (HPA: tissue IHC; UniProt P43007 topology).
| Compartment and epitope orientation | UniProt annotates 8 membrane segments plus cytoplasmic and extracellular regions (UniProt P43007 topology). Interpret membrane-associated signal in that context; the supplied record gives no antibody epitope, so it cannot predict which side an antibody recognises or how retrieval affects it. |
| Cell-specific reference pattern | HPA calls squamous epithelial cells in cervix, esophagus and oral mucosa High, but adipocytes and liver cholangiocytes Not detected (HPA: tissue IHC). Use the named cell populations as reference points; a tissue-wide positive or negative label would overstate those observations. |
| Strength of IHC evidence | HPA labels the tissue IHC assessment Enhanced and reports medium staining–RNA consistency (HPA: tissue IHC). Antibody HPA034964 has Enhanced IHC validation; HPA034963 has no IHC status in the supplied antibody list (HPA: antibodies). Validation supports comparison with the pattern, not unquestioned assignment of every stained cell. |
| IF/ICC comparison — does its pattern change the IHC call? | HPA reports an Enhanced centrosome ICC-IF location (HPA: subcellular ICC-IF). That observation belongs to cultured-cell IF/ICC; it does not replace the cytoplasmic, often membranous tissue IHC reference pattern (HPA: tissue IHC). |
| Other annotated locations and forms | UniProt lists melanosome membrane identification by mass spectrometry and 2 isoforms (UniProt P43007 subcellular location; isoforms). Neither annotation establishes a distinct paraffin-section staining pattern or an isoform-specific readout for the supplied antibodies. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive structure is blank, and the study sections are blank. | The run may have failed; the blank result alone cannot establish absence where HPA reports High staining (HPA: pancreas; HPA: cerebral cortex). | Confirm the reference structure is present, then check the antibody's IHC validation, retrieval and detection steps against the established workflow (general IHC practice). No target-specific fixation sensitivity is reported in the supplied sources. |
| The entire section has diffuse chromogen, including cells expected to be unstained. | Excess background or endogenous detection activity can obscure a cell-specific result (general IHC practice); HPA reports some named cell populations as Not detected (HPA: tissue IHC). | Inspect a primary-omission control; check blocking, washes, antibody concentration and development time (general IHC practice). Score only a resolved pattern that matches the relevant cell population. |
| Staining is nuclear-only in both the test and known-positive tissue. | That distribution conflicts with the HPA tissue IHC profile and UniProt membrane annotation (HPA: tissue IHC; UniProt P43007 subcellular location). | Compare the primary-omission control and the validated antibody's reference image or pattern, if available (general IHC practice). Do not score nuclear-only deposit as the expected IHC result. |
| Adipocytes or cholangiocytes stain strongly despite little signal in the selected positive control. | HPA reports those specific populations as Not detected (HPA: adipocytes; HPA: cholangiocytes); antibody-dependent staining or detection background needs investigation (general IHC practice). | Verify cell identity and compare primary-omission and known-positive controls (general IHC practice). Avoid extending either HPA negative call to every cell in the tissue. |
| Cortex neuropil stains, but a reader expects every nearby cell body to stain equally. | The HPA High call names neuropil, a structure rather than all cortex cells (HPA: cerebral cortex). | Score the named structure separately and use HPA's cytoplasmic, often membranous tissue profile to describe any other cell staining (HPA: tissue IHC). |
| A centrosomal ICC-IF image appears inconsistent with a paraffin-section IHC result. | HPA reports centrosome localisation for ICC-IF and cytoplasmic, often membranous staining for tissue IHC (HPA: subcellular ICC-IF; tissue IHC). | Interpret this chromogenic paraffin section against the tissue IHC reference and its named cell populations (HPA: tissue IHC). Use the separate IF/ICC guide for IF-specific experimental decisions. |
Experiment evidence
IHC & IF images showing SLC1A4 staining
PA2186 · IHCAnti-SLC1A4 antibody, PA2186, IHC(P) IHC(P): Human Intestinal Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115902-SLC1A4/tissue/cerebral+cortex#imid_15342978
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000115902-SLC1A4/tissue/adrenal+gland#imid_15342946
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115902-SLC1A4/tissue/colon#imid_15339656
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115902-SLC1A4/tissue/liver#imid_15339657
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034963; green = SLC1A4, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034963; green = SLC1A4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC1A4 IHC & IF
🧪Run esophagus first and look for staining in squamous epithelial cells (HPA: High in esophageal squamous epithelial cells). Use adipose tissue as the negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the esophagus slide, adjacent stromal cells are candidate internal negatives and should show only background, but verify their status rather than assuming they lack SLC1A4 (HPA: High call applies to squamous epithelial cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC1A4 in A-431, U-251MG, U2OS, with annotated localisation: Centrosome (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a host- and clonality-matched isotype control, and a biological negative such as SLC1A4 knockout tissue or a validated peptide block (standard IHC practice). For chromogenic esophagus sections, quench endogenous peroxidase and check for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected PA2186 IHC(P) caption does not state a fixative (caption: fixative not stated). Antigen retrieval dependence for SLC1A4 is unreported, so optimize retrieval on paired positive and negative paraffin sections; the supplied evidence does not establish whether frozen sections or IF are easier (standard IHC practice; caption: IHC(P)). In esophagus, assess section edges for nonspecific chromogen staining before scoring the squamous epithelium (standard IHC practice; HPA: High in esophageal squamous epithelial cells).
HPA tissue IHC evidence for SLC1A4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC1A4 IHC Tips
Troubleshoot SLC1A4 staining in paraffin sections by checking retrieval, compartment, cell type and controls before interpreting chromogenic signal.
How should I retrieve SLC1A4 in paraffin sections with weak staining?
For SLC1A4 paraffin IHC, start with citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). Compare serial sections processed with the same antibody, detection chemistry and exposure to chromogen, using pancreatic exocrine glandular cells as a positive reference (HPA: High in pancreatic exocrine glandular cells). If signal remains weak, check deparaffinisation and heating uniformity before cautiously testing longer retrieval on adjacent sections (standard IHC practice). Score membrane-associated and cytoplasmic staining separately because both patterns occur in tissue IHC; diffuse staining alone does not establish successful retrieval (HPA: tissue IHC profile).
Could fixation explain variable SLC1A4 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected PA2186 tissue-IHC caption identifies paraffin sections but does not state the fixative (PA2186 caption). Record fixative, fixation duration and processing history for each block, then compare sections stained in the same run with identical retrieval and chromogen development (standard IHC practice). Include a reference section with documented strong staining, such as pancreatic exocrine glandular cells, to detect run-level failure (HPA: High in pancreatic exocrine glandular cells). If blocks differ, report the association with processing history without assigning the difference to a particular SLC1A4 epitope or modification (UniProt P43007: topology and modified residues).
Where should convincing SLC1A4 staining appear in tissue sections?
Assess cell borders and cytoplasm separately: SLC1A4 is annotated at the cell membrane, while tissue IHC shows general cytoplasmic expression, often with a membranous pattern (UniProt P43007: subcellular location; HPA: tissue IHC profile). Its 8 transmembrane segments make a membrane-associated pattern biologically plausible, but section orientation can make borders difficult to resolve (UniProt P43007: topology; standard IHC practice). Use a positive reference such as pancreatic exocrine glandular cells and compare adjacent negative-control sections before calling faint diffuse colour specific (HPA: High in pancreatic exocrine glandular cells; standard IHC practice). Treat a purely nuclear pattern as discordant with the supplied tissue and protein localisation evidence (HPA: tissue IHC profile; UniProt P43007: subcellular location).
How do epitope position and isoforms affect SLC1A4 IHC interpretation?
SLC1A4 has 2 annotated isoforms, but no antibody epitope or isoform coverage is supplied here (UniProt P43007: isoforms; PA2186 caption). Obtain the catalog antibody's immunogen or epitope information before claiming that a staining pattern represents either isoform (standard IHC practice). The protein has extracellular loops, cytoplasmic regions and 8 transmembrane segments; accessibility in processed sections may depend on where an epitope lies (UniProt P43007: topology; standard IHC practice). Glycosylation sites at 201 and 206 and phosphorylated residues near the cytoplasmic C terminus warrant care when comparing epitopes, but their effect on this antibody is untested (UniProt P43007: glycosylation and modified residues).
How should I follow up a chromogenic SLC1A4 result with IF?
For a separate IF experiment, multiplex SLC1A4 with a marker identifying the expected cell population, such as pancreatic exocrine glandular cells, and retain single-colour controls (HPA: High in pancreatic exocrine glandular cells; standard IF practice). Choose fluorophores and imaging channels after checking tissue autofluorescence in an unstained section (standard IF practice). If the antibody recognises a cytoplasmic epitope, test gentle permeabilisation; for an extracellular epitope, compare staining without permeabilisation, since its epitope is unspecified here (UniProt P43007: topology; PA2186 caption; standard IF practice). Compare any centrosomal IF signal with the tissue result cautiously: the reported centrosome location comes from cellular IF evidence, whereas tissue IHC often appears cytoplasmic with a membranous pattern (HPA: subcellular location; HPA: tissue IHC profile).
How can I reduce diffuse brown background without losing membrane signal?
Run a no-primary control and inspect whether brown colour follows tissue edges, damaged areas or endogenous pigment before adjusting the antibody step (standard IHC practice). Use an appropriate protein block, wash thoroughly, and block endogenous peroxidase when using peroxidase and DAB detection (standard chromogenic IHC practice). Titrate the catalog antibody and shorten DAB development on matched sections while preserving a positive reference such as pancreatic exocrine glandular cells (HPA: High in pancreatic exocrine glandular cells; standard IHC practice). Record cytoplasmic and membrane-associated staining independently, because both occur in SLC1A4 tissue IHC and aggressive background reduction can erase a weak border pattern (HPA: tissue IHC profile; standard IHC practice).
What should I score when comparing SLC1A4 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of positive cells and an intensity-based H-score for membrane-associated and cytoplasmic signal separately (HPA: tissue IHC profile; standard IHC practice). For sparse populations, report positive cells per mm² of viable tissue instead of raw counts (standard IHC practice). Normalise scores to the same tissue area or eligible cell population, with matched section thickness, staining run and chromogen development (standard IHC practice). Keep pancreatic exocrine glandular cells as a positive reference where available, and avoid pooling their scores with other cell populations (HPA: High in pancreatic exocrine glandular cells; standard IHC practice).
When is an apparent SLC1A4-positive area likely to be artefact?
Prefer reproducible cell-associated cytoplasmic or membrane-associated staining in the expected population over colour confined to cut edges or necrotic areas (HPA: tissue IHC profile; standard IHC practice). Pancreatic exocrine glandular cells provide a reported strong reference, while adipocytes are reported as not detected in HPA tissue IHC (HPA: High in pancreatic exocrine glandular cells; HPA: Not detected in adipocytes). Check a no-primary section for endogenous enzyme activity or pigment that could mimic DAB signal (standard chromogenic IHC practice). A purely nuclear pattern conflicts with the supplied membrane annotation and tissue profile; confirm any surprising compartment or cell type with an independent control before interpreting it as SLC1A4 (UniProt P43007: subcellular location; HPA: tissue IHC profile; standard IHC practice).
Boster reagents
Best SLC1A4 / Neutral amino acid transporter A IHC Antibodies
PA2186 has an IHC(P) image of human intestinal cancer tissue (PA2186 image caption). Human, Mouse and Rat are listed as reactive (PA2186 catalog reactivity).
Real IHC data
Anti-SLC1A4 Antibody ®
Cat # PA2186
PA2186 is listed for IHC in Human, Mouse and Rat (PA2186 catalog applications/reactivity). Its IHC(P) image shows human intestinal cancer tissue (PA2186 image caption).
Which to pick: Choose PA2186 for paraffin-section IHC: its own image caption documents IHC(P) on human intestinal cancer tissue, and its listed IHC concentration is 0.5–1 μg/ml (PA2186 image caption; PA2186 catalog IHC dilution). No listed SKU has IF/ICC as an application or an IF image, so there is no documented IF/ICC choice here (PA2186 catalog applications/IF images). PA2186 lists Human, Mouse and Rat reactivity, but its tissue image documents only Human; clonality and the fixative used for that image are unreported (PA2186 catalog reactivity/clone; PA2186 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P43007 (SATT_HUMAN, Neutral amino acid transporter A).
- Human Protein Atlas. SLC1A4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC1A4 subcellular location (ICC-IF): Localized to the centrosome..
- Human Protein Atlas. SLC1A4 antibody validation summary (2 antibodies).
- Divergent brain gene expression patterns associate with distinct cell-specific tau neuropathology traits in progressive supranuclear palsy. Acta neuropathologica 2018 — PMC6208732.
- DMNQ induces ferroptosis and augments the efficacy of anti-PD-L1 immunotherapy in gastric cancer via the STAT3/SLC1A4 axis to mediate cysteine metabolism reprogramming. Redox biology 2026 — PMC12870803.
- SLC1A4: A Powerful Prognostic Marker and Promising Therapeutic Target for HCC. Frontiers in oncology 2021 — PMC7991385.
- Skullcapflavone II Inhibits SLC1A4-Mediated L-Serine Uptake and Promotes Mitochondrial Damage in Gastric Cancer. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2025 — PMC12677694.
- PubMed PMID:8101838 — UniProt-cited evidence.
- PubMed PMID:8340364 — UniProt-cited evidence.
- PubMed PMID:7896285 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC1A4 (Neutral amino acid transporter A). Expected localisation Cytoplasmic, often membranous in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: PA2186 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PA2186); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SLC1A4, 0.5-1μg/ml (datasheet PA2186); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; extracellular vs cytoplasmic epitope matters (UniProt); Regulation: Tissue-dependent expression (HPA tissue IHC). Evidence: UniProt P43007, HPA tissue IHC · Enhanced, HPA ICC-IF · centrosome., Datasheet PA2186, PMC6208732, PMC7991385. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC1A4/immunohistochemistry-ihc-guide.