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SLC27A5 / Long-chain fatty acid transport protein 5 · IHC design guide
Design Immunohistochemistry for SLC27A5
Plan chromogenic IHC on paraffin sections using A30575 at 1:100–1:300 (datasheet: IHC dilution). Assess selective cytoplasmic staining in hepatocytes and score it against appropriate controls (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC27A5 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 | Hepatocyte cytoplasm (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic staining in hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Liver | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Endogenous liver peroxidase may raise DAB background (standard IHC practice) | |
| Regulation | Liver-enriched expression; other regulation unreported (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; cytoplasmic epitope coverage unverified (UniProt) |
Section 1
Recommended SLC27A5 IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol with the published SLC27A5 liver-section protocol (PMC10787101).
| Sample | Paraffin-embedded human liver carcinoma tissue; fixative not specified (datasheet A30575) |
| Fixation | Image fixative and duration unreported (datasheet A30575); 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-SLC27A5, 1:100 - 1:300 (datasheet A30575) |
| 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 | SLC27A5-positive staining in hepatocytes of liver (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in hepatocytes. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule; UniProt Q9Y2P5 topology).
Section 2
What Is the Expected SLC27A5 Staining Pattern?
SLC27A5 is predominantly expressed in liver (UniProt Q9Y2P5), and the expected IHC signal is selective, high cytoplasmic staining in hepatocytes (HPA: liver hepatocytes High; tissue IHC Enhanced). Its annotated locations include endoplasmic reticulum, microsomal and cell membranes; two transmembrane segments anchor the protein, while most of its sequence is cytoplasmic (UniProt Q9Y2P5 topology).
| Strong cytoplasmic staining in hepatocytes, with surrounding cells much less conspicuous. | This matches the observed liver pattern (HPA: selective cytoplasmic expression in hepatocytes; High). Score the hepatocyte signal and its distribution separately from background (general IHC practice). HPA's Enhanced reliability reflects consistency between staining and RNA data; it does not establish the specificity of every new staining run (HPA: tissue IHC Enhanced). |
| Predominantly nuclear staining, with little hepatocyte cytoplasmic signal. | A nuclear dominant pattern conflicts with the reported cytoplasmic liver staining (HPA: selective cytoplasmic expression in hepatocytes) and the annotated membrane locations (UniProt Q9Y2P5). Treat it as suspect until a no primary control and an independent IHC validated antibody help assess nonspecific detection (general IHC practice; HPA: multiple antibodies IHC Enhanced). |
| Strong staining in liver cells other than hepatocytes, or in a listed negative tissue. | This departs from the selective hepatocyte pattern (HPA: liver hepatocytes High). Adipocytes in adipose tissue and hematopoietic cells in bone marrow were not detected in the supplied tissue profile (HPA: Not detected). Cross reactivity or endogenous detection activity are possibilities to investigate, rather than diagnoses from appearance alone (general IHC practice). |
| Diffuse color over many compartments, with no clear hepatocyte contrast. | The expected signal is selective cytoplasmic hepatocyte staining (HPA: tissue IHC). Widespread haze is insufficient to call SLC27A5 positive; excess antibody, incomplete blocking or detection background can produce a similar appearance (general IHC practice). Compare a no primary control and inspect whether identifiable hepatocytes remain stronger than adjacent cells (general IHC practice). |
| No visible hepatocyte signal in an otherwise interpretable liver section. | This conflicts with the supplied high liver hepatocyte staining (HPA: High). First check tissue identity, staining controls and whether the IHC validated antibody's prescribed conditions were followed (general IHC practice). A negative slide alone cannot distinguish failed detection from a genuine difference in the specimen; HPA reports a reference tissue pattern, not a guarantee for every sample (HPA: tissue IHC). |
💡Expected SLC27A5 appearanceCall the result positive when hepatocytes show strong, selective cytoplasmic staining (HPA: liver hepatocytes High); nuclear dominant signal or comparable staining throughout listed negative tissue cells is suspect (HPA: selective cytoplasmic pattern; Not detected examples).
| Which tissue and cells set the reference pattern? | Liver hepatocytes provide the positive comparison (HPA: High; tissue enriched RNA). The supplied adipose tissue adipocytes, adrenal glandular cells and bone marrow hematopoietic cells were not detected (HPA: Not detected). Those findings are cell specific observations, not a claim that every cell in those organs lacks SLC27A5 (HPA: supplied tissue profile). |
| How should membrane localization affect scoring? | The IHC observation is cytoplasmic in hepatocytes (HPA: tissue IHC), while UniProt also annotates endoplasmic reticulum, microsomal and cell membranes (UniProt Q9Y2P5). Membrane accentuation within a hepatocyte can be assessed in that context, but surface staining alone does not establish antibody specificity (general IHC practice). |
| What does antibody validation support? | Three supplied antibodies have IHC Enhanced status: HPA007292, CAB056160 and CAB068239 (HPA: antibody validation). The tissue profile also has Enhanced reliability based on agreement with RNA expression (HPA: tissue IHC). These support the reference pattern; they do not validate an unlisted antibody, a particular dilution or a new specimen (HPA: supplied validation scope). |
| Can isoforms or processing explain a different pattern? | UniProt lists two isoforms and a full length chain with no annotated signal peptide, propeptide or glycosylation site (UniProt Q9Y2P5). The supplied record gives no antibody epitope or isoform specific staining data, so it cannot assign a changed IHC pattern to an isoform or processing event (UniProt Q9Y2P5; HPA: supplied antibody data). |
| What should IF/ICC show? | HPA summarizes the subcellular location as membrane but supplies no main location, ICC/IF image cell line or ICC validation for these antibodies (HPA: subcellular summary; antibody validation). An IHC hepatocyte pattern therefore gives no resolved IF/ICC staining prediction for a particular cell model (HPA: supplied IF/ICC data). |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver hepatocytes are blank. | The result disagrees with the reference positive pattern (HPA: liver hepatocytes High); tissue selection or an IHC detection step may be responsible (general IHC practice). | Confirm hepatocytes are present, review the IHC validated antibody's instructions, and run an appropriate positive control beside the sample (general IHC practice). |
| The no primary control also develops color. | Signal without primary antibody indicates background from the detection workflow, including possible endogenous enzyme activity in chromogenic IHC (general IHC practice). | Address the detection background using controls and the detection system's blocking procedure before interpreting SLC27A5 staining (general IHC practice). |
| Negative reference tissue cells stain as strongly as hepatocytes. | That conflicts with their supplied Not detected observations (HPA: adipose adipocytes; adrenal glandular cells; bone marrow hematopoietic cells). Nonspecific antibody binding remains possible (general IHC practice). | Verify cell identity and compare a no primary control; repeat with an independent IHC Enhanced antibody if available (general IHC practice; HPA: antibody validation). |
| Nuclei dominate the staining. | Nuclear dominant signal conflicts with the cytoplasmic hepatocyte pattern (HPA: tissue IHC) and annotated membrane locations (UniProt Q9Y2P5). | Recheck counterstain versus chromogen, then compare a no primary control and an independent IHC validated antibody before scoring nuclei as positive (general IHC practice). |
| Diffuse haze obscures hepatocyte boundaries. | Poor contrast prevents assessment of the selective cytoplasmic pattern (HPA: tissue IHC); antibody concentration, blocking or detection background may contribute (general IHC practice). | Use the antibody's IHC guidance to review dilution and blocking, then assess controls and hepatocyte to adjacent cell contrast (general IHC practice). |
| Two IHC validated antibodies give different liver patterns. | Their shared IHC Enhanced status does not establish equivalent epitopes or identical performance in every specimen (HPA: antibody validation; supplied data omit epitopes). | Compare each antibody's controls and staining conditions, document the discrepancy, and base interpretation on the reproducible hepatocyte pattern supported by the tissue reference (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing SLC27A5 staining
A30575 · IHCImmunohistochemistry analysis of paraffin-embedded human liver carcinoma tissue, using SLC27A5 Antibody. The picture on the right is blocked
A30575 · IFImmunofluorescence analysis of A549 cells, using SLC27A5 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000083807-SLC27A5/tissue/cerebral+cortex#imid_14920876
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000083807-SLC27A5/tissue/colon#imid_14921386
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000083807-SLC27A5/tissue/liver#imid_14921379
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000083807-SLC27A5/tissue/kidney#imid_14921380
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC27A5 IHC & IF
🧪Run liver first: hepatocytes should stain strongly (HPA: High in hepatocytes). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); non-hepatocyte cells on the liver slide should show only background staining if used as internal negatives, but their SLC27A5 status is not established by the supplied HPA row.
Positive control tissue: Liver (Hepatocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for SLC27A5; derive a cell-line control from the positive tissue's cell type (Hepatocytes) and confirm it by RNA or western blot first.
Technical controls: Include no-primary/secondary-only and host- and clonality-matched isotype controls, plus a knockout specimen or peptide-block control (selected-SKU tissue-IHC caption: peptide block). For chromogenic liver IHC, quench endogenous peroxidase and check for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not report a fixative (selected-SKU tissue-IHC caption). Optimize antigen retrieval empirically for paraffin sections; the supplied evidence does not establish that frozen sections or IF are easier (selected-SKU tissue-IHC caption: paraffin section). Liver pigment can complicate chromogenic scoring, while autofluorescence can complicate IF interpretation (standard IHC/IF practice).
HPA tissue IHC evidence for SLC27A5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended 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 | Endocrine 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 SLC27A5 IHC Tips
Troubleshoot chromogenic IHC for SLC27A5 in paraffin sections using hepatocyte staining, membrane topology and appropriate controls as guides (HPA tissue IHC; UniProt Q9Y2P5).
How should I retrieve SLC27A5 in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval specification). Allow sections to cool in buffer before washing, then compare a consistently processed liver control with the test sections (standard IHC practice; HPA tissue IHC: high in hepatocytes). If staining remains weak, test a longer retrieval time on adjacent sections while checking for tissue damage and increased background (standard IHC practice). Score the resulting cytoplasmic hepatocyte signal rather than treating stronger staining in any compartment as improved detection (HPA tissue IHC: selective cytoplasmic hepatocyte expression).
Could fixation explain weak SLC27A5 staining in my paraffin sections?
The selected antibody image shows paraffin-embedded human liver carcinoma, but its caption does not report a fixative (A30575 tissue-IHC caption). Target-specific fixation sensitivity is therefore unknown; neither the hepatocyte staining profile nor membrane topology establishes how fixation changes this antibody’s signal (HPA tissue IHC; UniProt Q9Y2P5 topology). Record fixative, fixation duration and processing for each specimen, and compare adjacent sections processed together with a liver control (standard IHC practice; HPA tissue IHC: high in hepatocytes). If fixation varies between groups, interpret intensity differences cautiously and establish staining consistency before making biological comparisons (standard IHC practice).
What staining pattern should count as SLC27A5-positive in liver IHC?
Expect predominantly cytoplasmic staining in hepatocytes, the cell population with high liver signal in the tissue profile (HPA tissue IHC: selective cytoplasmic expression; high in hepatocytes). SLC27A5 is associated with endoplasmic reticulum, microsomal and cell membranes, so a cytoplasmic pattern can include membrane-associated signal without requiring an exclusively surface outline (UniProt Q9Y2P5 subcellular annotation). Assess intact hepatocytes and their cytoplasm against the counterstain, using the same detection and viewing conditions across sections (standard IHC practice). Strong nuclear-only staining or staining concentrated in a different cell population should trigger review of morphology, background controls and antibody specificity before scoring (HPA tissue IHC; standard IHC practice).
Can this antibody distinguish SLC27A5 isoforms or reveal epitope masking?
SLC27A5 has 2 annotated isoforms, but the supplied antibody evidence does not map its epitope or establish isoform selectivity (UniProt Q9Y2P5 isoforms; A30575 tissue-IHC caption). Its annotated membrane spans are residues 31–51 and 56–76, with residues 77–690 on the cytoplasmic side (UniProt Q9Y2P5 topology). Compare retrieval conditions on adjacent sections if signal is weak, but do not interpret a change in staining as isoform recognition without epitope information (standard IHC practice). The caption reports inhibition by a synthesized peptide; that observation supports an epitope-dependent signal in that image but does not establish isoform discrimination (A30575 tissue-IHC caption).
How should I assess SLC27A5 by IF alongside chromogenic IHC?
Treat IF as a separate validation question: the selected antibody caption documents paraffin-section IHC, not IF performance (A30575 tissue-IHC caption). Multiplex SLC27A5 with an independently validated hepatocyte marker and check that target signal falls in the expected cell population (HPA tissue IHC: selective cytoplasmic hepatocyte expression; standard IF practice). Choose a far-red target channel, approximately 650–700 nm, after measuring tissue autofluorescence and checking channel bleed-through (standard IF practice). Because residues 1–30 and 77–690 face the cytoplasm, access to an epitope in either region requires suitable permeabilisation; the antibody’s actual epitope is unreported (UniProt Q9Y2P5 topology; A30575 tissue-IHC caption; standard IF practice).
How can I reduce diffuse or misleading SLC27A5 chromogenic background?
Inspect a no-primary control before changing antibody concentration; widespread chromogen there points toward the detection workflow rather than interpretable SLC27A5 staining (standard IHC practice). For peroxidase-based detection, use an endogenous peroxidase block, such as 3% hydrogen peroxide for 10 min, and verify that it reduces control-section signal (standard IHC practice). Apply an appropriate protein block, wash thoroughly and titrate the primary antibody against a liver control while preserving hepatocyte cytoplasmic staining (standard IHC practice; HPA tissue IHC: high in hepatocytes). The selected image includes peptide blocking, but that comparison alone does not exclude tissue background or unrelated cross-reactivity in other specimens (A30575 tissue-IHC caption; standard IHC practice).
How should I quantify SLC27A5 IHC across liver specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment as hepatocyte cytoplasm and exclude folds, necrosis and section edges before measuring signal (HPA tissue IHC: selective cytoplasmic hepatocyte expression; standard IHC practice). Report the percentage of positive hepatocytes and an intensity-weighted H-score from 0–300, using prespecified intensity categories and a consistent threshold (standard IHC scoring practice). Normalise counts to the number of evaluable hepatocytes, or area-based measurements to evaluable tissue area in mm², rather than total slide area (standard IHC practice). Keep retrieval, staining, imaging and scoring conditions aligned across batches, and include a liver control to track technical drift (page retrieval specification; HPA tissue IHC; standard IHC practice).
When is apparent SLC27A5 staining more likely to be artefact?
A credible positive pattern is cytoplasmic staining in intact hepatocytes, consistent with the tissue profile and the protein’s membrane-associated localisation (HPA tissue IHC: selective cytoplasmic hepatocyte expression; UniProt Q9Y2P5 subcellular annotation). Review isolated nuclear signal or dominant staining in the wrong cell population against morphology and controls before calling it SLC27A5 (HPA tissue IHC; standard IHC practice). Edge accentuation, folds and necrotic regions can create uneven chromogenic signal, so exclude them from interpretation and scoring (standard IHC practice). Signal in a no-primary control may reflect endogenous enzyme activity or detection background; assess that control alongside the stained section (standard IHC practice).
Boster reagents
Best SLC27A5 / Long-chain fatty acid transport protein 5 IHC Antibodies
A30575 has IHC data from paraffin-embedded human liver carcinoma tissue and IF data from A549 cells; its listed reactivity is human, mouse and rat (catalog: A30575 images and reactivity).
Real IHC data
Anti-SLC27A5/Fatp5 Antibody
Cat # A30575
A30575 will render with an IHC image of paraffin-embedded human liver carcinoma tissue with a peptide-blocked comparison (catalog: A30575 IHC caption). Its listed applications include IHC, IF and ICC, and its IF image shows A549 cells with a peptide-blocked comparison (catalog: A30575 applications and IF caption).
Which to pick: Choose A30575 for paraffin-section IHC: it is a rabbit polyclonal antibody listed for IHC, with an image from paraffin-embedded human liver carcinoma tissue (catalog: A30575 applications and IHC caption). The caption does not report the fixative (catalog: A30575 IHC caption). For IF/ICC or work across human, mouse and rat samples, A30575 is the listed option; its application list includes IF and ICC, its IF image shows A549 cells, and its stated reactivity covers those three species (catalog: A30575 applications, IF caption and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y2P5 (S27A5_HUMAN, Long-chain fatty acid transport protein 5).
- Human Protein Atlas. SLC27A5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLC27A5 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SLC27A5 antibody validation summary (3 antibodies).
- Loss of SLC27A5 Activates Hepatic Stellate Cells and Promotes Liver Fibrosis via Unconjugated Cholic Acid. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2024 — PMC10787101.
- Decreased SLC27A5 Suppresses Lipid Synthesis and Tyrosine Metabolism to Activate the Cell Cycle in Hepatocellular Carcinoma. Biomedicines 2022 — PMC8869743.
- PubMed PMID:10479480 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC27A5 (Long-chain fatty acid transport protein 5). Expected localisation Hepatocyte cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A30575 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver carcinoma tissue; fixative not specified (datasheet A30575); 2. Fixation: Image fixative and duration unreported (datasheet A30575); 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-SLC27A5, 1:100 - 1:300 (datasheet A30575); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Endogenous liver peroxidase may raise DAB background (standard IHC practice); Isoform / epitope: 2 isoforms; cytoplasmic epitope coverage unverified (UniProt); Regulation: Liver-enriched expression; other regulation unreported (HPA tissue IHC). Evidence: UniProt Q9Y2P5, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A30575, PMC10787101. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC27A5/immunohistochemistry-ihc-guide.