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SLC2A8 / Solute carrier family 2, facilitated glucose transporter member 8 · IHC design guide
Design Immunohistochemistry for SLC2A8
Plan SLC2A8 chromogenic IHC in paraffin sections using testis Leydig cells as a high-staining reference (HPA tissue IHC) and the catalog antibody at 1:25 (datasheet M06641). Interpret cytoplasmic staining cautiously because the tissue IHC profile has uncertain reliability and presumed off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC2A8 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 | Low cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | High in testis Leydig cells; low cytoplasmic in a few tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet M06641) | |
| Positive control | Testis | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Use formaldehyde-fixed paraffin sections (datasheet M06641). (selected-SKU IHC image M06641) | |
| Caveat | Presumed off-target binding makes tissue staining uncertain (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; epitope orientation matters across 12 transmembrane segments (UniProt) |
Section 1
Recommended SLC2A8 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M06641) is followed by a published mouse brain IHC protocol (PMC2480596).
| Sample | Formaldehyde-fixed, paraffin-embedded human testis tissue (datasheet M06641) |
| Fixation | Image formalin-fixed; duration unreported (datasheet M06641); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet M06641); 20 min, 95–100 °C (standard) |
| 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-SLC2A8, 1:25 (datasheet M06641) |
| Primary incubation | 1 hours at 37°C (datasheet M06641) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC2A8-positive staining in leydig cells of testis (HPA tissue IHC: High). HPA tissue profile: Low cytoplasmic expression in a few tissues, highly abundant in testis. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet M06641). Compare staining with Slc2a8 knockout brain sections as a specificity control (PMC2480596).
Section 2
What Is the Expected SLC2A8 Staining Pattern?
SLC2A8 is principally intracellular, on cytoplasmic vesicle membranes, but may move to the plasma membrane (UniProt Q9NY64 subcellular location). In paraffin-section IHC, the strongest reported staining is in testis Leydig cells; lower cytoplasmic staining occurs in a few other tissues (HPA tissue IHC). Interpret that pattern cautiously: HPA rates its tissue IHC evidence Uncertain because presumed off-target binding was observed and disregarded, pending external verification (HPA tissue IHC).
| Strong cytoplasmic, possibly granular staining in testis Leydig cells. | This fits the reported High Leydig-cell signal (HPA tissue IHC) and principally intracellular vesicle localization (UniProt Q9NY64). Chromogenic IHC may not resolve individual vesicles (general IHC practice); morphology alone cannot verify antibody specificity, particularly with HPA's Uncertain tissue-IHC rating (HPA tissue IHC). |
| A rim of membrane staining accompanies cytoplasmic staining. | A membrane component can be plausible because SLC2A8 may move between vesicles and the plasma membrane (UniProt Q9NY64 subcellular location). Interpret a membrane-only or predominantly nuclear pattern cautiously against the expected intracellular localization (UniProt Q9NY64; HPA subcellular ICC-IF). |
| Strong staining appears in a cell population HPA lists as Not detected. | For example, adipocytes in adipose tissue are listed as Not detected (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogen activity (general IHC practice), then compare cell morphology and a detection-only control; the HPA designation applies to the named cells, not every cell in that tissue (HPA tissue IHC). |
| Color spreads diffusely across cells, stroma, or the section. | A diffuse deposit is difficult to reconcile with the reported Leydig-cell enrichment and cytoplasmic pattern (HPA tissue IHC). Uneven detection reagent, inadequate blocking, or excessive antibody concentration can cause background (general IHC practice); a detection-only control helps locate the source. |
| No signal is visible in Leydig cells on the testis section. | HPA reports High Leydig-cell staining, but rates the tissue IHC evidence Uncertain (HPA tissue IHC). Check tissue preservation, retrieval, antibody incubation, and detection with appropriate controls (general IHC practice) before treating a blank section as evidence that SLC2A8 is absent. |
💡Expected SLC2A8 appearanceA plausible positive is High cytoplasmic staining in testis Leydig cells (HPA tissue IHC), consistent with mainly vesicular localization (UniProt Q9NY64; HPA subcellular ICC-IF); widespread diffuse or predominantly nuclear color should prompt artifact checks (general IHC practice).
| Tissue and cell selection | Testis Leydig cells provide the strongest reported IHC signal; hepatocytes, lung macrophages, and several other listed populations show Low staining (HPA tissue IHC). Assess the named cell population rather than judging an entire organ positive or negative (HPA tissue IHC). |
| Intracellular localization | SLC2A8 is principally intracellular and can cycle to the cell surface (UniProt Q9NY64 subcellular location). Expect a cytoplasmic component; surface staining alone is insufficient to establish specificity, while an absolute ban on membrane staining would conflict with the UniProt annotation. |
| Topology and epitope access | The protein has 12 transmembrane segments and both cytoplasmic and extracellular regions (UniProt Q9NY64 topology). The supplied record does not identify this antibody's epitope, so topology cannot determine which retrieval condition will work or predict a target-specific fixation effect. |
| Glycosylation | One glycosylation site is annotated at residue 349 (UniProt Q9NY64 glycosylation). Its presence alone does not establish whether this antibody recognizes a modified epitope or whether glycosylation changes chromogenic staining; interpret staining through controls rather than that annotation alone. |
| Antibody evidence | HPA011935 is rated Uncertain for tissue IHC; HPA reports presumed off-target binding and pending external verification (HPA tissue IHC; HPA antibodies). A matching distribution is supportive, but independent specificity evidence would strengthen a biological conclusion (general IHC practice). |
| IF/ICC Q: Does vesicular IF validate tissue IHC? | A: HPA reports approved vesicular ICC-IF localization and lists CACO-2, Hep-G2, and U2OS images (HPA subcellular ICC-IF; HPA antibodies). That supports an intracellular localization expectation; tissue IHC retains its separate Uncertain rating (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Leydig cells are blank, including on the intended positive section. | HPA reports High signal there, although its tissue IHC rating is Uncertain (HPA tissue IHC); a processing or detection failure is possible (general IHC practice). | Verify tissue morphology and the staining run's controls, then review deparaffinization, retrieval, primary-antibody incubation, and chromogen development (general IHC practice). Do not infer a specific retrieval condition from the supplied SLC2A8 record. |
| The entire section has brown haze. | Excess antibody, insufficient washing, or nonspecific detection can produce diffuse chromogenic background (general IHC practice). | Compare a detection-only control, inspect wash and blocking steps, and titrate the primary antibody using the selected positive and negative cell populations (general IHC practice; HPA tissue IHC). |
| Color appears even without primary antibody. | Endogenous enzyme activity or nonspecific detection chemistry can contribute signal in chromogenic IHC (general IHC practice). | Check the detection-only control and the blocking appropriate to the enzyme system; reassess the tissue pattern after controlling that background (general IHC practice). |
| Predominantly nuclear staining is reproducible. | The reported locations are vesicle and cell membranes, with HPA ICC-IF showing vesicles (UniProt Q9NY64; HPA subcellular ICC-IF); nuclear dominance lacks support in these sources. | Check the counterstain and detection-only control, then seek independent antibody or orthogonal evidence before calling nuclear color SLC2A8 (general IHC practice; HPA tissue IHC: Uncertain). |
| A listed negative cell population stains strongly. | For example, HPA lists adipose-tissue adipocytes as Not detected (HPA tissue IHC); cross-reactivity or staining artifact is possible, especially with Uncertain IHC reliability (HPA tissue IHC). | Confirm cell identity, compare the named negative cells with Leydig cells on matched runs, and investigate background with a detection-only control (HPA tissue IHC; general IHC practice). |
| Low-level staining in liver or lung is hard to score. | HPA assigns Low staining to hepatocytes and lung macrophages (HPA tissue IHC); faint chromogen can approach background (general IHC practice). | Score the specified cells against local background and controls, document intensity and distribution, and avoid calling weak diffuse color positive solely because an organ appears on HPA's Low list (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing SLC2A8 staining
M06641 · IHCM06641 staining SLC2A8 in human testis tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136856-SLC2A8/tissue/cerebral+cortex#imid_3316592
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136856-SLC2A8/tissue/colon#imid_3316510
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136856-SLC2A8/tissue/liver#imid_3316511
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136856-SLC2A8/tissue/kidney#imid_3316528
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA011935; green = SLC2A8, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA011935; green = SLC2A8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC2A8 IHC & IF
🧪Run testis first and require staining in Leydig cells (HPA: High in Leydig cells). Use adipose tissue as the negative tissue, with adipocytes lacking specific staining (HPA: Not detected in adipocytes); on the testis slide, internal negative cells should show counterstain without specific chromogen signal.
Positive control tissue: Testis (Leydig cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC2A8 in CACO-2, Hep-G2, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, an immunoglobulin control matched to the primary antibody’s host species and, if monoclonal, its isotype, and SLC2A8 knockout tissue as a biological negative. Check endogenous peroxidase and biotin background in testis when using chromogenic detection with a biotinylated secondary (selected-SKU IHC caption: biotinylated secondary).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU fixative field is unreported (selected-SKU caption metadata: fixative not stated). The IHC-P caption uses heat retrieval in citrate buffer at pH 6, but does not establish that retrieval is required (selected-SKU IHC caption: citrate retrieval); the supplied evidence does not establish whether frozen sections or IF are easier. IF offers a vesicular localization reference in CACO-2, Hep-G2 and U2OS (HPA: approved vesicular location and ICC-IF images); testis-specific artefacts are unreported.
HPA tissue IHC evidence for SLC2A8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Presumed off target binding observed and disregarded. Pending external verification.). 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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC2A8 IHC Tips
Use matched controls and compartment aware scoring to evaluate SLC2A8 staining in paraffin sections; tissue IHC reliability remains uncertain (HPA: Uncertain).
How should I adjust retrieval when SLC2A8 staining is weak?
Start with heat mediated antigen retrieval in citrate buffer at pH 6 (datasheet M06641). This condition accompanies SLC2A8 staining in human testis paraffin sections using the catalog antibody at 1:25 for 1 hour at 37°C (caption M06641). If staining is weak, compare modest changes in heating time on adjacent sections while holding antibody incubation and chromogen development constant (standard IHC practice). Include testis with expected Leydig cell staining and a no primary control; increased diffuse signal or damaged morphology is not an improvement (HPA: High in Leydig cells; standard IHC practice).
Could fixation explain weak or patchy SLC2A8 staining?
SLC2A8 specific sensitivity to fixation is unknown from the supplied evidence, so a weak result alone cannot identify overfixation as its cause (supplied record: no fixation sensitivity data). Record fixative, duration, section thickness, and processing history for each paraffin block before comparing staining (standard IHC practice). Test sections processed alike with the same retrieval, antibody incubation, and chromogen development, including an expected positive testis control (HPA: High in Leydig cells; standard IHC practice). If processing histories differ, interpret intensity differences cautiously and assess tissue preservation and nonspecific staining alongside the target signal (standard IHC practice).
Should I expect membrane staining or intracellular puncta?
Prioritise intracellular, potentially punctate staining when examining SLC2A8: it is principally intracellular and is assigned to cytoplasmic vesicle membranes (UniProt Q9NY64: subcellular location). Vesicle localisation is also reported in cell imaging, while movement to the plasma membrane remains possible (HPA: Vesicles; UniProt Q9NY64: subcellular location). In testis, assess the expected Leydig cell compartment against surrounding cells rather than treating every stained profile as equivalent (HPA: High in Leydig cells). Compare staining with a no primary control and intact morphology; isolated surface rims, pigment, or precipitate require scrutiny before assignment to transporter localisation (standard IHC practice).
How does epitope position affect interpretation of SLC2A8 IHC?
The supplied record lists a 477 amino acid chain, 12 transmembrane segments, and no annotated isoforms, but does not identify this antibody's epitope (UniProt Q9NY64: processing, topology, isoforms; supplied caption M06641). Therefore, do not assume that a negative section excludes SLC2A8 expression or that retrieval exposes every possible epitope equally (standard IHC practice). UniProt places glycosylation at residue 349, within an extracellular region, although its relevance to this antibody is unknown (UniProt Q9NY64: glycosylation and topology). If epitope mapping becomes available, check its membrane facing side and compare staining under controlled retrieval conditions (standard IHC practice).
How can IF help check an ambiguous chromogenic SLC2A8 pattern?
Use IF as a separate localisation check alongside IHC, pairing SLC2A8 with a validated Leydig cell marker in testis (HPA: High in Leydig cells; standard IF practice). Choose well separated fluorophores and favour a far red channel if tissue autofluorescence obscures shorter wavelengths; include single stain and no primary controls (standard IF practice). SLC2A8 is principally intracellular on vesicle membranes, so access to an intracellular epitope may require permeabilisation (UniProt Q9NY64: subcellular location; standard IF practice). Because this antibody's epitope is unspecified, verify its membrane facing side before interpreting a negative IF result, and compare compartment patterns rather than equating fluorescence intensity with DAB intensity (supplied caption M06641; standard IF practice).
What should I troubleshoot when DAB appears throughout the section?
First compare the stained slide with a no primary control and inspect whether colour follows tissue structures or deposits outside cells (standard IHC practice). The selected testis staining used 3% BSA blocking for 30 minutes and a biotinylated secondary antibody (caption M06641). For a peroxidase and DAB workflow, check endogenous peroxidase blocking, secondary antibody specificity, and chromogen development time; a biotin based system also warrants an endogenous biotin control (standard IHC practice; caption M06641: biotinylated secondary). Reassess concentration against the reported 1:25 primary dilution, and accept changes only when expected Leydig cell staining remains distinguishable from background (caption M06641; HPA: High in Leydig cells).
How should I score SLC2A8 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: testis Leydig cells are the strongest reported tissue IHC finding, while SLC2A8 is principally intracellular (HPA: High in Leydig cells; UniProt Q9NY64: subcellular location). For chromogenic staining, record the percentage of viable positive cells and intensity on a consistent 0–3 scale, then calculate an H score from 0–300 (standard IHC practice). Normalise to the number of evaluable target cells or viable tissue area, and keep acquisition and threshold settings fixed across slides (standard IHC practice). Report compartment specific results and control performance because the HPA tissue staining assessment is uncertain (HPA: Uncertain; standard IHC practice).
How can I distinguish genuine SLC2A8 signal from artefact?
A credible testis result should show reproducible staining in Leydig cells with an intracellular pattern compatible with vesicle localisation (HPA: High in Leydig cells; HPA: Vesicles). Plasma membrane staining can be biologically possible, but a uniform rim alone needs independent support because SLC2A8 is principally intracellular (UniProt Q9NY64: subcellular location). Examine section edges, necrotic regions, pigment, and no primary controls for staining that tracks damage or persists without primary antibody (standard IHC practice). Check endogenous peroxidase contribution when using DAB, and avoid treating a positive slide as definitive specificity evidence because the tissue IHC reliability assessment is uncertain (standard IHC practice; HPA: Uncertain).
Boster reagents
Best SLC2A8 / Solute carrier family 2, facilitated glucose transporter member 8 IHC Antibodies
One IHC-validated anti-SLC2A8 antibody has a human testis paraffin-section image (catalog image caption); listed reactivity covers human, mouse, and rat (catalog reactivity).
Real IHC data
Anti-SLC2A8 Antibody (Center)
Cat # M06641
M06641 has an IHC-P image from human testis paraffin sections (catalog image caption). Its listed applications include IHC-P and flow cytometry, with human, mouse, and rat reactivity (catalog applications and reactivity).
Which to pick: Choose M06641 for tissue IHC: its own image documents staining in human testis paraffin sections, with citrate-buffer heat retrieval and a 1:25 primary dilution (M06641 image caption). No SKU in this payload lists IF/ICC validation, so there is no validated IF/ICC pick (catalog applications). M06641 is the cross-species candidate because it is a rabbit polyclonal antibody listed as reactive with human, mouse, and rat; its supplied IHC image documents human tissue only (catalog antibody details; M06641 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NY64 (SL2A8_HUMAN, Solute carrier family 2, facilitated glucose transporter member 8).
- Human Protein Atlas. SLC2A8 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. SLC2A8 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. SLC2A8 antibody validation summary (1 antibodies).
- Targeted disruption of Slc2a8 (GLUT8) reduces motility and mitochondrial potential of spermatozoa. Molecular membrane biology 2008 — PMC2557070.
- Deletion of glucose transporter GLUT8 in mice increases locomotor activity. Behavior genetics 2008 — PMC2480596.
- PubMed PMID:10821868 — UniProt-cited evidence.
- PubMed PMID:10671487 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC2A8 (Solute carrier family 2, facilitated glucose transporter member 8). Expected localisation Low cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet M06641). Validated IHC antibody: M06641 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Leydig cells); negative Adipose tissue. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded human testis tissue (datasheet M06641); 2. Fixation: Image formalin-fixed; duration unreported (datasheet M06641); 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 retrieval: Citrate pH 6 (datasheet M06641); 20 min, 95–100 °C (standard); 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-SLC2A8, 1:25 (datasheet M06641); 9. Primary incubation: 1 hours at 37°C (datasheet M06641). Watch for: Fixation: Use formaldehyde-fixed paraffin sections (datasheet M06641). (selected-SKU IHC image M06641); Caveat: Presumed off-target binding makes tissue staining uncertain (HPA tissue IHC); Isoform / epitope: No isoforms annotated; epitope orientation matters across 12 transmembrane segments (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt Q9NY64, HPA tissue IHC · Uncertain, HPA ICC-IF · vesicles., Datasheet M06641, PMC2480596. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC2A8/immunohistochemistry-ihc-guide.