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AKR1B10 / Aldo-keto reductase family 1 member B10 · IHC design guide
Design Immunohistochemistry for AKR1B10
Plan AKR1B10 paraffin IHC with the catalog antibody at 0.5–1 μg/mL (datasheet A02976). Assess cytoplasmic staining in intestinal enterocytes and stomach glandular cells, using HPA tissue staining as a reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AKR1B10 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 | Cytoplasm in gastrointestinal epithelium (HPA tissue IHC) | |
| Staining pattern | Enterocyte and glandular-cell cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A02976) | |
| Positive control | Small intestine+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) | |
| Caveat | Secretion may separate tissue RNA and protein patterns (UniProt; HPA tissue IHC) | |
| Regulation | Staining-linked regulation is unreported (UniProt) | |
| Isoform / epitope | No isoforms annotated; one chain spans residues 1–316 (UniProt) |
Section 1
Recommended AKR1B10 IHC & IF Protocols
Compare the catalog antibody's citrate pH 6 IHC-P protocol (datasheet A02976) with four published AKR1B10 chromogenic IHC protocols (PMC5011670; PMC6201704; PMC8305663; PMC11663324).
| Sample | Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02976) |
| Fixation | Image fixative and duration unreported (datasheet A02976); 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, 20 min (datasheet A02976) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02976) |
| Primary antibody | Rabbit anti-AKR1B10, 0.5-1μg/ml (datasheet A02976) |
| Primary incubation | Overnight at 4 °C (datasheet A02976) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02976) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AKR1B10-positive staining in enterocytes - Microvilli of small intestine (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in gastrointestinal tract and gall bladder. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet A02976); use Tris pH 8.5 when reproducing the automated endometrial protocol (PMC8305663).
Section 2
What Is the Expected AKR1B10 Staining Pattern?
In paraffin sections, AKR1B10 should stain mainly the cytoplasm of gastrointestinal epithelial cells, with high staining reported in small intestinal enterocytes and stomach glandular cells (HPA: tissue IHC, Enhanced). The protein has no transmembrane segment, although UniProt also lists lysosomal localization and lysosome-mediated secretion (UniProt O60218: topology and subcellular location). HPA cautions that its tissue profile may include proteins from more than one gene (HPA: tissue IHC reliability note).
| Clear cytoplasmic staining in small intestinal enterocytes or stomach glandular cells. | This matches the reported high-staining cell types and the gastrointestinal cytoplasmic profile (HPA: tissue IHC). Assess the named cells against adjacent tissue before calling the section positive; staining elsewhere in the section does not establish the expected pattern. |
| Predominantly nuclear staining, or a sharply outlined cell surface without a convincing cytoplasmic component. | Treat a nuclear-only pattern as suspect because HPA reports cytoplasmic tissue staining (HPA: tissue IHC). A surface signal alone is also insufficient: HPA reports additional plasma membrane localization in ICC-IF, with a multiple-gene antibody caution (HPA: subcellular). |
| Strong staining mainly in cells outside the reported epithelial populations. | The cell distribution does not match the reported positive populations (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic detection activity (general IHC practice); identify the cells and check detection controls before assigning the signal to AKR1B10. |
| Diffuse color across epithelium, stroma and blank areas, obscuring cell boundaries. | This is background rather than a readable cell-restricted pattern (general IHC practice). Compare with a no-primary control and inspect blocking, washes and detection conditions; the HPA tissue profile cannot identify which workflow step produced the background. |
| No interpretable staining in small intestine or stomach. | Absence in these reported high-staining tissues calls the run into question (HPA: tissue IHC). First confirm that the expected enterocytes or glandular cells are present, then review the catalog antibody’s IHC-P instructions and the run controls (general IHC practice). |
💡Expected AKR1B10 appearanceCall a positive result when small intestinal enterocytes or stomach glandular cells show clear, predominantly cytoplasmic staining at the reported High level (HPA: tissue IHC); isolated nuclear color or diffuse color without cell definition is suspect (HPA: tissue IHC; general IHC practice).
| Tissue and cell selection | Small intestinal enterocytes and stomach glandular cells are reported High; appendix and colon enterocyte microvilli, duodenal enterocytes and gallbladder glandular cells are Medium (HPA: tissue IHC). Compare the same cell population when judging intensity. |
| Localization and secretion | UniProt lists lysosomal localization and secretion through a lysosome-mediated pathway, with no transmembrane segment (UniProt O60218: subcellular location and topology). These annotations do not make extracellular color alone a diagnostic positive IHC pattern; HPA describes the tissue pattern as cytoplasmic (HPA: tissue IHC). |
| Strength and limits of the tissue evidence | The tissue IHC profile is rated Enhanced for agreement between staining and RNA, but HPA cautions that the antibodies can target proteins from more than one gene and that secretion complicates RNA–protein agreement (HPA: tissue IHC reliability note). Interpret staining in its tissue and cell context. |
| Adrenal gland discrepancy | UniProt describes AKR1B10 as highly expressed in adrenal gland (UniProt O60218: tissue specificity), whereas HPA reports adrenal glandular cells as Not detected by tissue IHC (HPA: tissue IHC). Do not use adrenal staining alone to define an expected positive or a failed run. |
| IF/ICC Q: What localization is reported? | A: Mainly cytosol, with additional plasma membrane localization (HPA: subcellular, supported). HPA cautions that these observations rely on antibodies targeting proteins from multiple genes (HPA: subcellular); they do not establish an exclusive AKR1B10 membrane pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive small intestine or stomach has no signal. | The expected cells may be absent from the section, or the staining run may have failed (HPA: tissue IHC; general IHC practice). | Locate enterocytes or glandular cells on the counterstained section, confirm that run controls worked, then follow the catalog antibody’s IHC-P instructions for retrieval and dilution (general IHC practice). No AKR1B10-specific retrieval sensitivity is established here. |
| Nuclear staining dominates while cytoplasm is faint. | The distribution conflicts with HPA’s cytoplasmic tissue profile (HPA: tissue IHC); the image alone cannot identify its cause. | Compare the same cell types in a reported high-staining tissue and inspect a no-primary control. Reassess the result with an independently validated antibody if the nuclear pattern persists (general IHC practice). |
| Color appears in unexpected cells or in a negative comparison tissue. | Possible cross-reactivity or endogenous detection activity needs checking (general IHC practice). HPA cautions that its antibodies may recognize proteins from more than one gene (HPA: tissue IHC reliability note). | Record the stained cell type, compare it with the HPA tissue profile, and use appropriate no-primary and detection controls before interpreting the color as target-specific (general IHC practice). |
| Uniform haze makes epithelial cells hard to score. | Widespread background can reflect blocking, washing or chromogenic detection conditions (general IHC practice); HPA’s tissue observations do not establish a target-specific cause. | Check the no-primary control, then review blocking, washes and detector conditions for the run. Score only where cell boundaries and the cytoplasmic pattern remain interpretable (general IHC practice; HPA: tissue IHC profile). |
| Adrenal gland is unstained despite the UniProt expression description. | The supplied sources differ: UniProt reports high adrenal expression, while HPA reports adrenal glandular cells as not detected by IHC (UniProt O60218: tissue specificity; HPA: tissue IHC). | Use HPA’s high-staining small intestine or stomach cell populations to judge the IHC run, and report the adrenal result without treating either source as proof of an assay failure. |
| A section shows only extracellular deposits or a thin cell-edge signal. | UniProt reports secretion and HPA reports additional plasma membrane localization in ICC-IF, but HPA’s tissue IHC pattern is cytoplasmic (UniProt O60218: subcellular location; HPA: subcellular and tissue IHC). | Look for convincing cytoplasmic staining in the expected epithelial cells, compare controls, and avoid scoring the isolated deposit or edge signal as a definitive AKR1B10-positive tissue pattern. |
Experiment evidence
IHC & IF images showing AKR1B10 staining
A02976 · IHCIHC analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in paraffin-embedded section of human intestinal cancer t
A02976 · IHCIHC analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in paraffin-embedded section of human liver cancer tissue
A02976 · IFIF analysis of AKR1B10 using anti-AKR1B10 antibody (A02976). AKR1B10 was detected in an immunocytochemical section of A549 cells. Enzyme ant
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/cerebral+cortex#imid_18963205
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/duodenum#imid_18963114
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/colon#imid_18963115
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198074-AKR1B10/tissue/liver#imid_18748143
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA020280; green = AKR1B10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA020280; green = AKR1B10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AKR1B10 IHC & IF
🧪Run small intestine first; enterocytes should stain (HPA: High in small intestine, Enterocytes - Microvilli). Use adipose tissue adipocytes as a negative tissue (HPA: Not detected in adipocytes); non-enterocyte cells on the small-intestine slide should be checked for background, but are not a validated negative population.
Positive control tissue: Small intestine (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AKR1B10 in A-549, Hep-G2, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, an isotype-matched rabbit IgG control, and AKR1B10 knockout material or immunizing-peptide competition if available (caption: rabbit primary antibody; standard IHC control practice). In small intestine, quench endogenous peroxidase and check for endogenous biotin signal with the streptavidin-biotin/DAB detection used in the example (caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A02976 paraffin-section caption does not state a fixative (caption: fixative not stated). Heat retrieval in citrate buffer at pH 6 for 20 minutes is a supported IHC starting condition, but retrieval dependence is unreported (caption: citrate retrieval, pH 6, 20 minutes). The supplied evidence does not establish whether frozen sections or IF are easier; endogenous biotin can create misleading intestinal staining with the example’s streptavidin-biotin detection (caption: SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for AKR1B10
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Small intestine | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Duodenum | Enterocytes | 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 → |
| 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 AKR1B10 IHC Tips
Troubleshoot AKR1B10 staining in paraffin sections by checking retrieval, cell type, compartment, controls, and scoring before interpreting signal.
How should I retrieve AKR1B10 in paraffin sections when staining is weak?
Use heat-mediated citrate pH 6 antigen retrieval for AKR1B10 paraffin-section IHC (datasheet A02976). The selected tissue image used 20 minutes of retrieval before incubation with the catalog antibody, providing a documented starting condition for this assay (caption A02976). If signal is weak, compare retrieved and unretrieved sections while keeping antibody concentration and detection conditions matched; assess whether staining increases in the expected epithelial cells rather than across the whole section (caption A02976; HPA tissue IHC; standard IHC practice). Record heating time and cooling conditions for each run, since inconsistent retrieval can make sections difficult to compare (standard IHC practice).
Can I attribute weak AKR1B10 staining to the tissue fixative?
The selected image documents a paraffin-embedded section but does not state its fixative, so target-specific fixation sensitivity is unknown (caption A02976). Do not infer fixation tolerance from the reported tissue distribution, absence of a transmembrane segment, or annotated acetyllysines at positions 125 and 263 (HPA tissue IHC; UniProt O60218 topology and modified residues). For a fixation comparison, process matched samples consistently, then apply the documented citrate pH 6 retrieval and identical staining conditions (datasheet A02976; standard IHC practice). Include a known positive tissue section in each run so processing failure can be distinguished from genuinely low expression in the sample under study (HPA tissue IHC; standard IHC practice).
Which AKR1B10 staining pattern is plausible in tissue sections?
Assess epithelial cytoplasmic staining first: tissue IHC reports cytoplasmic expression in the gastrointestinal tract and gallbladder, with high staining in small-intestinal enterocytes and stomach glandular cells (HPA tissue IHC). Supported cell imaging places the protein mainly in the cytosol and additionally at the plasma membrane, although that subcellular evidence carries a cross-gene antibody caution (HPA subcellular). A peripheral pattern can therefore be evaluated alongside cytoplasmic signal, while a purely nuclear pattern warrants a specificity check (HPA subcellular; standard IHC practice). UniProt also annotates lysosomal localisation and secretion through a lysosome-mediated pathway, so extracellular signal needs cell-level controls before assignment to its source (UniProt O60218 subcellular location).
Could an AKR1B10 epitope or related protein explain discordant IHC staining?
UniProt lists 0 isoforms, one annotated chain spanning residues 1–316, and no transmembrane segment for AKR1B10 (UniProt O60218 processing, isoforms, and topology). Those annotations do not identify the catalog antibody's epitope or establish whether fixation and citrate retrieval preserve its binding site (UniProt O60218; datasheet A02976). The HPA tissue reliability note cautions that its antibodies target proteins from more than one gene, so agreement with those images alone cannot prove AKR1B10 specificity (HPA tissue IHC). When staining conflicts with expected cell patterns, compare independently validated reagents or an orthogonal expression readout on matched material before assigning the signal to AKR1B10 (HPA tissue IHC; standard IHC practice).
How can I check an AKR1B10 pattern by multiplex immunofluorescence?
Pair AKR1B10 with a validated marker for the expected epithelial cell type, then inspect whether the signals occupy the same cells in the relevant tissue compartment (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores after checking unstained tissue for autofluorescence, and include single-label controls to assess channel bleed-through (standard IF practice). For an epitope on the cytosolic side of a membrane, test permeabilisation against a matched unpermeabilised sample; the antibody epitope is unspecified, while AKR1B10 has no transmembrane segment (UniProt O60218 topology; standard IF practice). Compare cytosolic and peripheral signal with the supported subcellular pattern, bearing in mind its cross-gene antibody caution (HPA subcellular).
What should I check when AKR1B10 DAB staining appears diffuse?
First compare a no-primary control with the stained section, then inspect whether deposit follows epithelial cytoplasm or indiscriminately covers stroma, lumina, and damaged areas (HPA tissue IHC; standard IHC practice). The selected image used 10% goat-serum blocking, 1 µg/ml primary antibody overnight at 4°C, a biotinylated secondary, and DAB development (caption A02976). If background persists, review washing, antibody concentration, endogenous peroxidase blocking, and possible endogenous biotin signal in the chosen detection workflow (caption A02976; standard IHC practice). Keep exposure to chromogen consistent across comparison sections, since overdevelopment can obscure the cell boundaries needed to judge AKR1B10 localisation (standard IHC practice).
How should I quantify heterogeneous AKR1B10 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored epithelial compartment before counting, using the expected gastrointestinal and gallbladder cytoplasmic pattern to guide annotation (HPA tissue IHC). Report the percentage of positive eligible cells and an intensity-weighted H-score, applying the same thresholds and scoring rules to every section (standard IHC practice). Normalise counts to the number of eligible epithelial cells, or normalise stained area to the annotated epithelial area, rather than total section area when tissue composition varies (standard IHC practice). Exclude folds, necrotic regions, and section edges using prespecified criteria, and record control-section performance so apparent differences can be checked against staining-run variation (standard IHC practice).
When is an unexpected AKR1B10 signal likely to be artefact?
A credible positive pattern should identify plausible cells and compartments: HPA reports gastrointestinal epithelial cytoplasmic staining, while supported subcellular data place AKR1B10 mainly in the cytosol (HPA tissue IHC; HPA subcellular). Treat isolated nuclear signal, uniform staining across unrelated cell types, or staining concentrated at section edges and necrotic regions as reasons to investigate artefact (HPA tissue IHC; standard IHC practice). Check a no-primary control and endogenous peroxidase blocking when interpreting DAB deposits, particularly because the selected workflow uses chromogenic development (caption A02976; standard IHC practice). Remember that HPA flags cross-gene antibody recognition and possible secretion, so localisation or agreement with RNA alone cannot establish protein identity (HPA tissue IHC).
Boster reagents
Best AKR1B10 / Aldo-keto reductase family 1 member B10 IHC Antibodies
The IHC-validated antibody has images from human paraffin-embedded intestinal and liver cancer sections and an IF image from A549 cells (catalog image captions; catalog reactivity: Human).
Real IHC data
Anti-AKR1B10 Antibody ®
Cat # A02976
A02976 has IHC images from human paraffin-embedded intestinal and liver cancer sections (A02976 IHC captions). A02976 also has an IF image from A549 cells and lists IF and ICC among its applications (A02976 IF caption; catalog applications).
Which to pick: Choose A02976 for paraffin-section IHC because its own IHC captions document staining in human intestinal and liver cancer sections; the fixative is unreported (A02976 IHC captions). Choose A02976 for IF/ICC because those applications are listed and its IF caption documents staining in A549 cells (catalog applications; A02976 IF caption). No cross-species option is supported: A02976 lists Human reactivity only (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O60218 (AK1BA_HUMAN, Aldo-keto reductase family 1 member B10).
- Human Protein Atlas. AKR1B10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AKR1B10 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the plasma membrane. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. AKR1B10 antibody validation summary (3 antibodies).
- Pretreatment AKR1B10 expression predicts the risk of hepatocellular carcinoma development after hepatitis C virus eradication. World journal of gastroenterology 2016 — PMC5011670.
- Immunohistochemistry Detects Increased Expression of Aldo-Keto Reductase Family 1 Member B10 (AKR1B10) in Early-Stage Hepatocellular Carcinoma. Medical science monitor : international medical journal of experimental and clinical research 2018 — PMC6201704.
- AKR1B1 and AKR1B10 as Prognostic Biomarkers of Endometrioid Endometrial Carcinomas. Cancers 2021 — PMC8305663.
- Aldo-keto reductase family 1 member B10 prevents esophageal squamous cell carcinoma from reactive carbonyl species-induced cell death and promotes its progression. Cancer cell international 2024 — PMC11663324.
- PubMed PMID:9565553 — UniProt-cited evidence.
- PubMed PMID:9765596 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AKR1B10 (Aldo-keto reductase family 1 member B10). Expected localisation Cytoplasm in gastrointestinal epithelium (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A02976). Validated IHC antibody: A02976 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Small intestine (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human intestinal cancer tissues; fixative not specified (datasheet A02976); 2. Fixation: Image fixative and duration unreported (datasheet A02976); 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 , 20 min (datasheet A02976); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02976); 8. Primary antibody: Rabbit anti-AKR1B10, 0.5-1μg/ml (datasheet A02976); 9. Primary incubation: Overnight at 4 °C (datasheet A02976). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secretion may separate tissue RNA and protein patterns (UniProt; HPA tissue IHC); Isoform / epitope: No isoforms annotated; one chain spans residues 1–316 (UniProt); Regulation: Staining-linked regulation is unreported (UniProt). Evidence: UniProt O60218, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the plasma…, Datasheet A02976, PMC5011670, PMC6201704. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AKR1B10/immunohistochemistry-ihc-guide.