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SLC25A23 / Mitochondrial adenyl nucleotide antiporter SLC25A23 · IHC design guide
Design Immunohistochemistry for SLC25A23
Plan paraffin IHC around granular CNS cytoplasmic staining (HPA tissue IHC) and the protein’s inner mitochondrial membrane location (UniProt). Use caudate or hippocampus neuronal cells as staining references (HPA tissue IHC), and consult the catalog antibody’s paraffin-section workflow (datasheet A08997-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLC25A23 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 | Granular cytoplasm in CNS (HPA tissue IHC); inner mitochondrial membrane (UniProt) | |
| Staining pattern | Granular neuronal cytoplasm in caudate and hippocampus (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08997-1) | |
| Positive control | Breast+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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08997-1) | |
| Caveat | Antibody staining and RNA show low consistency (HPA tissue IHC) | |
| Regulation | Brain-enriched RNA (HPA tissue RNA) | |
| Isoform / epitope | 4 isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended SLC25A23 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet: A08997-1). One published IHC protocol reports SLC25A23 staining in colon cancer tissue (PMC9569204: methods).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A08997-1) |
| Fixation | Image fixative and duration unreported (datasheet A08997-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A08997-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08997-1) |
| Primary antibody | Rabbit anti-SLC25A23, 2-5 μg/ml (datasheet A08997-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08997-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08997-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLC25A23-positive staining in glandular cells of breast (HPA tissue IHC: Medium). HPA tissue profile: Granular cytoplasmic expression in CNS with additional cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A08997-1); the published colon cancer protocol does not specify retrieval conditions (PMC9569204: methods).
Section 2
What Is the Expected SLC25A23 Staining Pattern?
SLC25A23 is an inner mitochondrial membrane protein with six transmembrane segments (UniProt Q9BV35 topology). In paraffin sections, expect granular cytoplasmic staining, particularly in CNS neuronal cells and several glandular or tubular cell populations (HPA tissue IHC: granular CNS staining; Medium in caudate and hippocampal neurons, breast glandular cells and kidney tubules). HPA rates tissue IHC as Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Granular cytoplasmic staining in hippocampal or caudate neurons, with an appropriate nuclear counterstain. | This fits the reported CNS pattern and Medium neuronal staining (HPA tissue IHC: hippocampus and caudate). Its cytoplasmic position is consistent with mitochondrial localization (UniProt Q9BV35; HPA subcellular ICC-IF). Chromogenic IHC alone does not resolve the inner mitochondrial membrane (general IHC practice). |
| Predominantly nuclear, cell-surface or extracellular staining, without a granular cytoplasmic component. | Those compartments conflict with the reported mitochondrial location (UniProt Q9BV35 topology; HPA subcellular ICC-IF). Treat the pattern as suspect and compare it with an expected-positive section and a detection control before assigning it to SLC25A23 (general IHC practice). |
| Strong signal in a cell population HPA lists as Not detected, such as adipocytes. | The result conflicts with the reported adipocyte observation (HPA tissue IHC: adipocytes Not detected). Possible explanations include antibody cross-reactivity or endogenous chromogenic detection activity (general IHC practice); the discordance alone cannot identify which cause applies, especially given HPA's low staining–RNA consistency (HPA tissue IHC reliability). |
| Diffuse color over cells and tissue spaces, obscuring cell boundaries and granules. | This is difficult to score as the reported granular cytoplasmic pattern (HPA tissue IHC profile). Widespread background can arise from detection chemistry, inadequate blocking or nonspecific antibody binding (general IHC practice); evaluate it with a primary-antibody omission control (general IHC practice). |
| No staining in hippocampal neurons or kidney tubule cells on a test section. | HPA reports Medium staining in these respective cell populations (HPA tissue IHC: hippocampus; kidney). Check tissue preservation, the antibody's IHC-P instructions and the detection system (general IHC practice). A blank section alone does not establish absent SLC25A23 expression (general IHC practice). |
💡Expected SLC25A23 appearanceCall a positive result when relevant neurons or glandular or tubular cells show discernible granular cytoplasmic staining around Medium intensity in HPA's listed examples; dominant nuclear or extracellular color is suspect (HPA tissue IHC: profile and listed Medium cell populations; UniProt Q9BV35 localization).
| Subcellular location and topology | SLC25A23 has 6 membrane-spanning segments in the mitochondrial inner membrane (UniProt Q9BV35 topology). Interpret chromogenic granules at the cytoplasmic level; do not claim that light microscopy resolves membrane topology (general IHC practice). |
| Tissue and cell selection | HPA reports Medium staining in caudate and hippocampal neurons, breast glandular cells and kidney tubule cells, while adipocytes are Not detected (HPA tissue IHC). Score the named cell populations rather than treating every cell in a tissue as equivalent (general IHC practice). |
| Evidence strength | The tissue IHC assessment is Approved, with low consistency between antibody staining and RNA expression; GTEx data supports IHC (HPA tissue IHC reliability). The listed patterns are useful comparators, but a discordant section needs investigation rather than an automatic biological conclusion (general IHC practice). |
| IF/ICC question: should the IF result look identical to chromogenic IHC? | No exact visual match is implied: HPA reports supported mitochondrial localization by ICC-IF, while tissue IHC reports granular cytoplasmic staining (HPA subcellular ICC-IF; HPA tissue IHC). The modalities support a compartment-level comparison, not identical image texture (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive neuronal or tubular cells are blank. | An assay or detection failure is possible; those cells are reported at Medium intensity (HPA tissue IHC: hippocampus, caudate and kidney). | Check the antibody's IHC-P instructions, section integrity and chromogenic detection controls, then compare a separate reported-positive section (general IHC practice; HPA tissue IHC: listed Medium cells). |
| Signal is mainly nuclear or along cell surfaces. | That distribution is inconsistent with mitochondrial localization (UniProt Q9BV35; HPA subcellular ICC-IF). Nonspecific staining or interpretation of background is possible (general IHC practice). | Recheck morphology and the primary-antibody omission control; score only convincing cytoplasmic granules in the relevant cells (general IHC practice; HPA tissue IHC profile). |
| Broad brown background masks the expected granules. | Nonspecific binding or chromogenic detection background may obscure the reported granular cytoplasmic pattern (general IHC practice; HPA tissue IHC profile). | Inspect the omission control and review blocking, washing and detection conditions under the IHC-P workflow (general IHC practice). |
| Cells listed as Not detected appear strongly positive. | Cross-reactivity or endogenous chromogenic activity is possible (general IHC practice); the observation conflicts with the relevant HPA cell-level entry (HPA tissue IHC). | Check the omission control and inspect the specific cell type, then compare reported-positive tissue before interpreting the discrepancy (general IHC practice; HPA tissue IHC: listed Medium cells). |
| CNS regions give different staining results. | HPA lists Medium staining in caudate and hippocampal neurons but Not detected in cerebellar granular-layer cells (HPA tissue IHC); region and cell identity can therefore change the comparison. | Record the region and scored cell population explicitly, and compare each with its matching HPA entry (HPA tissue IHC). |
| A tissue result disagrees with an RNA-based expectation. | HPA reports low consistency between antibody staining and RNA expression, despite GTEx support for its IHC assessment (HPA tissue IHC reliability). | Document the observed cell-level pattern and assay controls; avoid converting RNA abundance alone into an expected IHC intensity (general IHC practice; HPA tissue IHC reliability). |
Experiment evidence
IHC & IF images showing SLC25A23 staining
A08997-1 · IHCIHC analysis of SLC25A23 using anti-SLC25A23 antibody (A08997-1). SLC25A23 was detected in a paraffin-embedded section of mouse brain tissue
A08997-1 · IHCIHC analysis of SLC25A23 using anti-SLC25A23 antibody (A08997-1). SLC25A23 was detected in a paraffin-embedded section of rat brain tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000125648-SLC25A23/tissue/cerebral+cortex#imid_14440984
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000125648-SLC25A23/tissue/colon#imid_14440910
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000125648-SLC25A23/tissue/liver#imid_14440911
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000125648-SLC25A23/tissue/kidney#imid_14440904
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA009675; green = SLC25A23, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA009675; green = SLC25A23, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLC25A23 IHC & IF
🧪Run hippocampus first: neuronal cells should show Medium staining (HPA: hippocampus, neuronal cells, Medium). Run cerebellum as the comparison, scoring cells in its granular layer where staining was not detected (HPA: cerebellum, cells in granular layer, Not detected); on the hippocampus slide, adjacent non-neuronal cells should remain near background, but their target-negative status is unverified (HPA: hippocampal result specifies neuronal cells).
Positive control tissue: Breast (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLC25A23 in CACO-2, PC-3, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control; and, if available, a matched SLC25A23 knockout specimen or validated immunogen-peptide competition control (selected IHC caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase for HRP/DAB detection, and check unstained brain sections for autofluorescence if using IF (selected IHC caption: HRP/DAB; standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A08997-1 paraffin-section caption does not state a fixative (selected IHC caption: fixative not stated). The caption uses heat-mediated EDTA pH 8.0 retrieval, but does not establish that retrieval is required; the supplied evidence does not show whether frozen sections or IF/ICC are easier (selected IHC caption: EDTA pH 8.0). For IF/ICC, mitochondrial localization is supported, while brain lipofuscin can complicate fluorescence interpretation (HPA subcellular: Mitochondria, supported; standard IF practice).
HPA tissue IHC evidence for SLC25A23
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. GTEx data supports IHC.). 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 |
|---|---|---|---|---|
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | 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 → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLC25A23 IHC Tips
Use the catalog antibody’s paraffin-section conditions as a starting point, then evaluate compartment, cell type and controls before scoring SLC25A23 staining.
What retrieval should I start with if SLC25A23 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA buffer at pH 8.0 (datasheet A08997-1). The selected mouse brain paraffin-section example used that retrieval, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A08997-1). If signal remains weak, compare retrieval heating and cooling conditions on adjacent sections while keeping antibody concentration and DAB development consistent (standard IHC practice). Score improvement by clearer cytoplasmic granularity in the expected cells, while checking that tissue structure and the no-primary control remain acceptable (HPA: granular cytoplasmic CNS expression; standard IHC practice).
How should I troubleshoot fixation-related loss of SLC25A23 staining?
The selected paraffin-section caption does not state its fixative, and target-specific fixation sensitivity is unknown (datasheet A08997-1). Compare sections with documented fixation histories under the same EDTA pH 8.0 retrieval and primary-antibody conditions before attributing a weak result to fixation (datasheet A08997-1; standard IHC practice). Include a consistently processed positive-reference section and a no-primary control in each comparison (standard IHC practice). Record fixation duration, tissue thickness, retrieval conditions and staining intensity together; differences between specimens alone cannot establish a fixation effect when their processing and cellular composition also differ (standard IHC practice).
What staining pattern should I expect for SLC25A23 in tissue sections?
Expect a cytoplasmic, potentially granular pattern at light-microscope resolution because SLC25A23 resides in the mitochondrial inner membrane (UniProt Q9BV35 localisation; HPA: granular cytoplasmic CNS expression). The selected paraffin-section example demonstrates staining in mouse brain, while HPA reports medium staining in caudate and hippocampal neuronal cells (datasheet A08997-1; HPA tissue IHC). Compare signal within morphologically identified cells and include an adjacent section with a mitochondrial marker if compartment assignment matters (standard IHC practice). Diffuse nuclear staining alone is difficult to reconcile with the reported mitochondrial location and warrants review of controls, counterstain and section quality (UniProt Q9BV35 localisation; standard IHC practice).
Could epitope position or alternative splicing explain inconsistent SLC25A23 staining?
SLC25A23 has 4 reported isoforms, but the supplied antibody caption does not identify its bound epitope or establish isoform coverage (UniProt Q9BV35 isoforms; datasheet A08997-1). The protein has 6 transmembrane segments, with residues 1–188 assigned to the intermembrane side and several intervening loops assigned to the matrix side (UniProt Q9BV35 topology). Request the catalog antibody’s immunogen or epitope information before interpreting a negative section as absence of every isoform (standard IHC practice). If staining varies across similarly processed sections, compare retrieval response and an independently validated antibody where available, documenting which epitopes each reagent can detect (standard IHC practice).
How can I check SLC25A23 localisation by multiplex IF after tissue IHC?
For the separate IF/ICC application, pair SLC25A23 with a neuronal marker when examining CNS cells and with a mitochondrial marker to assess subcellular overlap (HPA: neuronal staining in caudate and hippocampus; HPA subcellular: mitochondria supported). Choose a fluorophore channel with low tissue autofluorescence and examine single-stain and no-primary controls before interpreting overlap (standard IF practice). Titrate permeabilisation against the antibody’s mapped epitope: the 1–188 region faces the intermembrane space, whereas several other loops face the matrix (UniProt Q9BV35 topology; standard IF practice). The supplied paraffin-section IHC caption gives no IF/ICC fixation or permeabilisation conditions for this antibody, so establish those conditions independently (datasheet A08997-1).
How do I reduce diffuse or misleading DAB background?
Begin with a no-primary control, a peroxidase block and matched DAB development to identify background from the detection workflow (standard chromogenic IHC practice). The selected paraffin-section example used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A08997-1). If diffuse staining persists, titrate primary antibody, strengthen appropriate blocking and washing, and shorten chromogen development one variable at a time (standard IHC practice). Preserve any granular cytoplasmic signal while checking edges, damaged areas and no-primary sections for staining that tracks tissue handling rather than cells (HPA: granular cytoplasmic CNS expression; standard IHC practice).
How should I quantify heterogeneous SLC25A23 staining across brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before measuring signal, because HPA reports medium staining in caudate and hippocampal neuronal cells but no detection in cerebellar granular-layer cells (HPA tissue IHC). Within each region, record the percentage of positive target cells and an intensity score, then calculate an H-score from those cell-level categories if intensity is reproducible (standard IHC practice). Normalise positive counts to the total number of eligible cells, or stained area to the measured tissue area in mm², using the same rule across sections (standard IHC practice). Exclude damaged tissue and set thresholds using controls stained in the same run (standard IHC practice).
How can I distinguish true SLC25A23 signal from staining artefact?
Give greatest weight to reproducible granular cytoplasmic staining in anatomically identified cells, consistent with the mitochondrial inner-membrane assignment and reported CNS pattern (UniProt Q9BV35 localisation; HPA tissue IHC). The selected mouse brain paraffin-section example supports use of the catalog antibody in that setting, but does not by itself identify every stained cell or establish fixation conditions (datasheet A08997-1). Investigate predominantly nuclear signal, section-edge accumulation, necrotic areas or staining shared by the no-primary control before calling a positive result (UniProt Q9BV35 localisation; standard IHC practice). Endogenous peroxidase can confound DAB interpretation, so compare blocked controls and repeat suspicious regions on intact sections (standard chromogenic IHC practice).
Boster reagents
Best SLC25A23 / Mitochondrial adenyl nucleotide antiporter SLC25A23 IHC Antibodies
The catalog antibody has paraffin-section IHC images from mouse and rat brain and human stomach (catalog image captions); no IF/ICC images are supplied (catalog payload).
Real IHC data
Anti-SLC25A23 Antibody ®
Cat # A08997-1
A08997-1 has paraffin-section IHC images from mouse and rat brain and human stomach (catalog image captions). Its listed applications include IHC, while IF/ICC is unlisted and has no image evidence (catalog applications; catalog image captions).
Which to pick: Choose A08997-1 for paraffin-section tissue IHC: its captions document EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, but do not report the fixative (catalog image captions). For work across species, it lists human, mouse and rat reactivity, with IHC images for all three; clonality is unreported (catalog reactivity; catalog image captions; catalog payload). No SKU in this payload is validated for IF/ICC, so an IF/ICC choice cannot be supported here (catalog applications; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BV35 (SCMC3_HUMAN, Mitochondrial adenyl nucleotide antiporter SLC25A23).
- Human Protein Atlas. SLC25A23 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SLC25A23 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. SLC25A23 antibody validation summary (2 antibodies).
- Comprehensive Analysis and Validation of Solute Carrier Family 25 (SLC25) and Its Correlation with Immune Infiltration in Pan-Cancer. BioMed research international 2022 — PMC9569204.
- PubMed PMID:15054102 — UniProt-cited evidence.
- PubMed PMID:15123600 — UniProt-cited evidence.
- PubMed PMID:15716113 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLC25A23 (Mitochondrial adenyl nucleotide antiporter SLC25A23). Expected localisation Granular cytoplasm in CNS (HPA tissue IHC); inner mitochondrial membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08997-1). Validated IHC antibody: A08997-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A08997-1); 2. Fixation: Image fixative and duration unreported (datasheet A08997-1); 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: EDTA pH 8.0 (datasheet A08997-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08997-1); 8. Primary antibody: Rabbit anti-SLC25A23, 2-5 μg/ml (datasheet A08997-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08997-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08997-1); Caveat: Antibody staining and RNA show low consistency (HPA tissue IHC); Isoform / epitope: 4 isoforms; check epitope coverage across variants (UniProt); Regulation: Brain-enriched RNA (HPA tissue RNA). Evidence: UniProt Q9BV35, HPA tissue IHC · Approved, HPA ICC-IF · mitochondria., Datasheet A08997-1, PMC9569204. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLC25A23/immunohistochemistry-ihc-guide.