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ALDOB / Fructose-bisphosphate aldolase B · IHC design guide
Design Immunohistochemistry for ALDOB
Plan chromogenic ALDOB IHC in paraffin sections using 0.5–1 μg/mL of the catalog antibody (datasheet A03893-2). Compare cytoplasmic staining in renal tubules and hepatocytes with adipose tissue adipocytes as a negative control (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ALDOB 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 | Selective cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic renal tubules, hepatocytes and intestinal endocrine cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03893-2) | |
| Positive control | Kidney+3 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Endogenous biotin may mimic ALDOB signal with SABC detection (datasheet A03893-2; standard IHC practice) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one chain spans residues 2–364 (UniProt) |
Section 1
Recommended ALDOB IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published ALDOB staining protocols from three articles (PMC9834807; PMC6606928; PMC5065259).
| Sample | Paraffin-embedded human liver cancer tissues; fixative not specified (datasheet A03893-2) |
| Fixation | Image fixative and duration unreported (datasheet A03893-2); 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 A03893-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03893-2) |
| Primary antibody | Rabbit anti-ALDOB, 0.5-1μg/ml (datasheet A03893-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03893-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03893-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ALDOB-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in renal tubules, hepatocytes, small intestine and duodenum. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A03893-2); the gastric tissue protocol also uses citrate at pH 6 (PMC5065259: Methods).
Section 2
What Is the Expected ALDOB Staining Pattern?
ALDOB is expected mainly in the cytoplasm and cytosol, with additional centrosome and centriolar satellite localisation reported by UniProt (UniProt P05062 subcellular location). In tissue IHC, look for selective cytoplasmic staining in renal tubule cells and hepatocytes, with staining also reported in small intestine and duodenum (HPA: tissue IHC profile). HPA rates the tissue staining evidence Enhanced, citing consistency between antibody staining and RNA expression (HPA: reliability). ALDOB has no transmembrane segment (UniProt P05062 topology).
| Strong cytoplasmic staining in renal tubule cells or hepatocytes. | This matches HPA's High staining in both cell populations and UniProt's cytoplasmic localisation (HPA: Kidney and Liver IHC; UniProt P05062 subcellular location). Compare the stained cells with the section's morphology before scoring intensity; the expected pattern is selective across cell types (HPA: tissue IHC profile). |
| Predominantly nuclear staining or a continuous cell-surface rim. | These dominant patterns conflict with the reported cytoplasmic distribution and absence of a transmembrane segment (HPA: tissue IHC profile; UniProt P05062 topology). Treat them as suspect and check controls and staining conditions (general IHC practice). UniProt also reports centrosome and centriolar satellite localisation, so an isolated cytoplasmic punctum alone is not proof of artefact (UniProt P05062 subcellular location). |
| Strong chromogen in cells expected to be unstained, such as adipocytes or bone marrow hematopoietic cells. | HPA reports ALDOB as Not detected in those specified cells (HPA: Adipose tissue and Bone marrow IHC). Consider antibody cross-reactivity or, if the detection system permits it, endogenous detection activity (general IHC practice). A stained negative cell is a reason to investigate, not by itself a diagnosis of which mechanism occurred. |
| Diffuse haze across cells, stroma, and tissue edges without selective cytoplasmic contrast. | The slide no longer resembles HPA's selective tissue pattern (HPA: tissue IHC profile). Review blocking, antibody concentration, washes, and detection background with appropriate controls (general IHC practice). Background alone cannot establish ALDOB expression in an unexpected cell population. |
| No detectable signal in renal tubules and hepatocytes. | Those are useful known-positive reference cells because HPA reports High staining in each (HPA: Kidney and Liver IHC). First confirm that tissue, detection, and counterstain are interpretable; then review antibody dilution and antigen retrieval using the antibody's IHC-P guidance (general IHC practice). No ALDOB-specific fixation sensitivity is supplied. |
💡Expected ALDOB appearanceA convincing positive is selective, predominantly cytoplasmic chromogen in renal tubule cells or hepatocytes at HPA-reported High staining levels; small-intestine endocrine cells are also reported High and duodenal endocrine cells Medium (HPA: tissue IHC). Dominant nuclear staining or broad staining of HPA-negative cells is suspect (UniProt P05062 subcellular location; HPA: negative tissue IHC).
| Which compartment should guide scoring? | Score cytoplasmic signal against cell morphology: HPA describes selective cytoplasmic tissue staining, and UniProt lists cytosol plus centrosome and centriolar satellites (HPA: tissue IHC profile; UniProt P05062 subcellular location). UniProt reports no transmembrane segment, so a dominant surface-rim pattern warrants review (UniProt P05062 topology). |
| Which tissues give useful contrasts? | Kidney tubule cells and hepatocytes are reported High; small-intestine endocrine cells are High, and duodenal endocrine cells Medium (HPA: tissue IHC). Adipocytes and bone marrow hematopoietic cells are reported Not detected (HPA: tissue IHC). Use the named cell populations when comparing positive and negative areas. |
| How strong is the tissue-pattern evidence? | HPA labels the tissue IHC reliability Enhanced and lists three antibodies with Enhanced IHC status: HPA002198, HPA073201, and CAB020827 (HPA: reliability; HPA: antibodies). That supports the reported tissue pattern; it does not specify performance for an unlisted catalog antibody or a different staining workflow. |
| What can this section conclude about IF/ICC? | HPA supplies no main ICC-IF location and lists no cell lines with ICC-IF images (HPA: subcellular record). UniProt reports cytosolic and centrosome-related localisation (UniProt P05062 subcellular location), but the supplied HPA record cannot validate an IF appearance. Consult the separate IF/ICC guide for that application. |
| What is known about retrieval and fixation? | No ALDOB-specific retrieval condition or fixation-effect evidence is supplied by UniProt or HPA (supplied UniProt P05062 and HPA records). Record the retrieval condition used and compare it with the antibody's IHC-P instructions when troubleshooting (general IHC practice); do not infer an ALDOB fixation effect from tissue intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive-control liver or kidney section has no cytoplasmic stain. | The result conflicts with HPA's High hepatocyte and renal tubule staining (HPA: Liver and Kidney IHC); the failed step cannot be identified from that observation alone. | Verify tissue identity and staining controls, then review antibody dilution, retrieval, and detection according to the antibody's IHC-P instructions (general IHC practice). No target-specific fixation explanation is established here. |
| The slide shows strong nuclear signal with little cytoplasmic contrast. | A dominant nuclear pattern does not match HPA's selective cytoplasmic tissue profile or UniProt's listed cytosolic localisation (HPA: tissue IHC profile; UniProt P05062 subcellular location). | Check the negative control and detection background, then compare the staining with a known-positive liver or kidney section (general IHC practice; HPA: Liver and Kidney IHC). Do not score nuclear signal alone as the expected ALDOB pattern. |
| Adipocytes or bone marrow hematopoietic cells stain strongly. | HPA reports ALDOB as Not detected in these cells (HPA: Adipose tissue and Bone marrow IHC). Cross-reactivity or endogenous activity of the chosen detection system are possible explanations, not established diagnoses (general IHC practice). | Compare a no-primary control with the test slide; address any system-specific endogenous activity and reassess antibody concentration and specificity (general IHC practice). Keep the interpretation tied to the named cells rather than the whole tissue. |
| Chromogen forms a widespread haze that obscures cell boundaries. | This prevents assessment of HPA's selective cytoplasmic pattern (HPA: tissue IHC profile). Excess background can arise from antibody concentration, insufficient blocking or washing, or the detection system (general IHC practice). | Inspect the no-primary control and optimise blocking, dilution, washing, and detection conditions one variable at a time (general IHC practice). Rescore only when cellular localisation and the counterstained tissue are readable. |
| A cell-surface rim dominates the staining. | UniProt reports no ALDOB transmembrane segment and places it in cytosolic and centrosome-related compartments (UniProt P05062 topology and subcellular location). The rim is therefore a poor match for the expected dominant signal. | Inspect tissue edges and the negative control for nonspecific deposition, and compare with a known-positive section showing cytoplasmic staining (general IHC practice; HPA: tissue IHC profile). Do not infer membrane localisation from this IHC result alone. |
| IF/ICC shows an unexpected pattern when compared with the IHC slide. | The supplied HPA subcellular record has no main ICC-IF location or cell-line images, so it provides no IF pattern for direct validation (HPA: subcellular record). The tissue IHC profile describes a separate application (HPA: tissue IHC). | Use the separate IF/ICC guide and application-appropriate controls to assess that result (general IF practice). For this IHC section, judge the chromogenic slide by its tissue cell type and cytoplasmic pattern (HPA: tissue IHC profile). |
Experiment evidence
IHC & IF images showing ALDOB staining
A03893-2 · IHCIHC analysis of ALDOB using anti-ALDOB antibody (A03893-2). ALDOB was detected in paraffin-embedded section of human liver cancer tissues. H
A03893-2 · IHCIHC analysis of ALDOB using anti-ALDOB antibody (A03893-2). ALDOB was detected in paraffin-embedded section of rat liver tissues. Heat media
A03893-2 · IFIF analysis of Aldolase/ALDOB using anti-Aldolase/ALDOB antibody (A03893-2). Aldolase/ALDOB was detected in immunocytochemical section of A4
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136872-ALDOB/tissue/cerebral+cortex#imid_1011800
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000136872-ALDOB/tissue/small+intestine#imid_1011693
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136872-ALDOB/tissue/colon#imid_1011718
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136872-ALDOB/tissue/liver#imid_1011727
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ALDOB IHC & IF
🧪Run liver first: hepatocytes should show cytoplasmic staining (HPA: High in hepatocytes; UniProt P05062: cytosolic localisation). Use adrenal gland as the negative tissue, assessing its glandular cells (HPA: Not detected in glandular cells); on the liver slide, non-hepatocyte areas should show only counterstain or background, but do not assume they are validated ALDOB-negative cells.
Positive control tissue: Kidney (Cells in tubules, 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 ALDOB; derive a cell-line control from the positive tissue's cell type (Cells in tubules) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, a matched rabbit IgG isotype control, and ALDOB-knockout material as a biological negative if available (caption: rabbit primary; standard IHC practice). For liver, quench endogenous peroxidase and control endogenous biotin signal when using the caption’s biotin-based SABC/DAB detection (caption: biotinylated secondary, SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03893-2 tissue-IHC caption does not state the fixative (caption: fixative not stated). The caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish that retrieval is required; the supplied evidence does not establish whether frozen sections or IF/ICC would be easier (caption: retrieval conditions; HPA subcellular: no ICC-IF image cell lines). In liver, endogenous peroxidase or biotin may contribute background with the reported SABC/DAB method (caption: SABC/DAB; standard IHC practice).
HPA tissue IHC evidence for ALDOB
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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Small intestine | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | 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 → |
| 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 ALDOB IHC Tips
Troubleshoot ALDOB staining in paraffin sections by checking retrieval, cell type, cytoplasmic localisation and controls before interpreting intensity.
How should I retrieve ALDOB in paraffin sections when staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A03893-2). The catalog antibody produced ALDOB staining in a paraffin-embedded human liver cancer section under these conditions, although the caption does not state its fixative (datasheet A03893-2). If staining is weak, check heating uniformity and section adhesion, then compare a modestly longer retrieval with the documented condition on adjacent sections (standard IHC practice). Keep antibody concentration and detection constant during that comparison; the documented primary concentration is 1 μg/ml with overnight incubation at 4°C (datasheet A03893-2).
Could fixation explain weak or uneven ALDOB staining?
Target-specific sensitivity of ALDOB staining to fixation is unknown: the catalog caption identifies a paraffin-embedded section but does not report the fixative (datasheet A03893-2). Record the fixative, fixation duration and section age for each specimen, and compare sections processed together before attributing differences to ALDOB abundance (standard IHC practice). If signal varies across a section, inspect morphology and compare tissue centers with edges for uneven processing or staining (standard IHC practice). Repeat the documented citrate pH 6, 20-minute retrieval consistently while investigating fixation; that retrieval is documented, whereas an ALDOB-specific fixation effect is not (datasheet A03893-2).
Where should convincing ALDOB staining appear in tissue?
Expect predominantly cytoplasmic staining: ALDOB is annotated in the cytosol and at the microtubule organizing center, centrosome and centriolar satellite (UniProt P05062 localisation). The tissue profile describes selective cytoplasmic expression in renal tubules, hepatocytes, small intestine and duodenum (HPA tissue IHC profile). Assess cell identity alongside compartment: cells in kidney tubules and liver hepatocytes are reported as high, while duodenal endocrine cells are medium (HPA tissue IHC). Predominantly nuclear or membrane outlining staining should trigger a review of morphology, controls and detection background before it is scored as ALDOB (UniProt P05062 localisation; standard IHC practice).
How can I assess whether the antibody detects an ALDOB epitope specifically?
The supplied record lists 0 isoforms and no transmembrane segment for ALDOB, so an isoform-specific or membrane-facing staining claim has no support here (UniProt P05062 record). It lists 14 modified residues, including phosphorylation at positions 36, 39 and 89, but the antibody epitope is unspecified (UniProt P05062 modified residues; datasheet A03893-2). Compare staining with the expected cytoplasmic pattern and include a no-primary control when investigating unexplained signal (HPA tissue IHC profile; standard IHC practice). Do not attribute retrieval sensitivity or staining differences to a particular modification without epitope information and direct experimental evidence (UniProt P05062 modified residues; datasheet A03893-2).
How should I investigate ALDOB by immunofluorescence alongside the IHC result?
Treat IF as a separate validation exercise: the supplied subcellular record lists no ICC/IF images, and the catalog tissue caption documents chromogenic staining only (HPA subcellular; datasheet A03893-2). Multiplex ALDOB with an independently validated marker identifying the expected cell type, such as hepatocytes or renal tubular cells, and compare cellular overlap rather than brightness alone (HPA tissue IHC profile; standard IF practice). Choose fluorophores after checking tissue autofluorescence and include single-label and no-primary controls to assess bleed-through and background (standard IF practice). Because ALDOB is cytosolic and lacks a transmembrane segment, establish sufficient permeabilisation for intracellular antibody access while preserving morphology (UniProt P05062 localisation and topology; standard IF practice).
What should I check when DAB staining is widespread or granular?
Run a no-primary control through the same detection sequence to identify signal from the secondary reagent, chromogen or tissue (standard IHC practice). The documented workflow used 10% goat serum blocking, a biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, and a streptavidin–biotin complex with DAB (datasheet A03893-2). Check endogenous peroxidase blocking and, for this detection scheme, assess endogenous biotin if background persists (standard chromogenic IHC practice). Compare suspicious deposits with the expected selective cytoplasmic pattern and avoid scoring diffuse extracellular or nuclear staining as ALDOB without corroboration (HPA tissue IHC profile; UniProt P05062 localisation).
How should I score ALDOB staining across different tissue specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the eligible cell population on morphology before scoring; HPA reports ALDOB in hepatocytes, renal tubular cells and intestinal cell populations (HPA tissue IHC). For each region, record the percentage of positive eligible cells and intensity, then use an H-score if intensity differences matter (standard IHC scoring practice). Normalize positive counts to the number of eligible cells, or normalize stained area to the measured tissue area in mm²; report the chosen denominator consistently (standard quantitative IHC practice). Exclude folds, detached tissue and necrotic regions using the same rules across specimens, and compare sections stained in the same run where possible (standard IHC practice).
How do I distinguish true ALDOB staining from artefact?
A credible result combines cytoplasmic staining with an expected cell population, such as hepatocytes or renal tubular cells (UniProt P05062 localisation; HPA tissue IHC). HPA reports ALDOB as not detected in adipocytes from adipose tissue and in adrenal glandular cells, making those populations useful context when reviewing unexpected staining (HPA tissue IHC). Examine section edges, folds and necrotic areas for patterns driven by processing, and compare suspicious signal with a no-primary control (standard IHC practice). Nuclear-dominant staining, diffuse extracellular DAB or signal attributable to endogenous peroxidase warrants investigation before calling a specimen ALDOB-positive (UniProt P05062 localisation; standard chromogenic IHC practice).
Boster reagents
Best ALDOB / Fructose-bisphosphate aldolase B IHC Antibodies
The IHC-validated antibody has paraffin-section images from human liver cancer and rat liver (A03893-2 IHC captions), plus IF/ICC images from A431, CACO-2 and U20S cells (A03893-2 IF captions).
Real IHC data
Anti-Aldolase/ALDOB Antibody ®
Cat # A03893-2
A03893-2 will render with its human liver cancer paraffin-section IHC image; a rat liver paraffin-section IHC image is also reported (A03893-2 IHC captions). The same SKU is listed for IHC and IF/ICC, with IF/ICC images from A431, CACO-2 and U20S cells (A03893-2 applications and IF captions).
Which to pick: Choose A03893-2 for paraffin-section IHC: its caption documents citrate retrieval at pH 6 for 20 minutes and primary antibody at 1 μg/ml (A03893-2 IHC caption); the fixative is unreported (A03893-2 IHC caption). Choose the same SKU for IF/ICC, with a listed primary concentration of 2 μg/ml; clonality is unreported (A03893-2 dilution data and clone field). For work across species, the catalog lists human, monkey, mouse and rat reactivity, while its IHC dilution entry specifically covers human, mouse and rat (A03893-2 reactivity and dilution data).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P05062 (ALDOB_HUMAN, Fructose-bisphosphate aldolase B).
- Human Protein Atlas. ALDOB tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ALDOB subcellular location (ICC-IF): Highest expression in SH-SY5Y: 22.7 nTPM.
- Human Protein Atlas. ALDOB antibody validation summary (3 antibodies).
- Prognostic values of ALDOB expression and (18)F-FDG PET/CT in hepatocellular carcinoma. Frontiers in oncology 2022 — PMC9834807.
- Dynamic bioenergetic alterations in colorectal adenomatous polyps and adenocarcinomas. EBioMedicine 2019 — PMC6606928.
- Downregulation of ALDOB is associated with poor prognosis of patients with gastric cancer. OncoTargets and therapy 2016 — PMC5065259.
- ALDOB represents a potential prognostic biomarker for patients with clear cell renal cell carcinoma. Translational andrology and urology 2023 — PMC10170270.
- PubMed PMID:6548561 — UniProt-cited evidence.
- PubMed PMID:2410860 — UniProt-cited evidence.
- PubMed PMID:6585824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ALDOB (Fructose-bisphosphate aldolase B). Expected localisation Selective cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03893-2). Validated IHC antibody: A03893-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissues; fixative not specified (datasheet A03893-2); 2. Fixation: Image fixative and duration unreported (datasheet A03893-2); 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 A03893-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03893-2); 8. Primary antibody: Rabbit anti-ALDOB, 0.5-1μg/ml (datasheet A03893-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03893-2). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Endogenous biotin may mimic ALDOB signal with SABC detection (datasheet A03893-2; standard IHC practice); Isoform / epitope: 0 annotated isoforms; one chain spans residues 2–364 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P05062, HPA tissue IHC · Enhanced, HPA cell-line RNA · Highest expression in SH-SY5Y: 22.7 nTPM, Datasheet A03893-2, PMC9834807, PMC6606928. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ALDOB/immunohistochemistry-ihc-guide.