This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ALDOB / Fructose-bisphosphate aldolase B · IHC design guide
Design Immunohistochemistry for ALDOB
Use liver hepatocytes or kidney tubules as positive tissue controls and expect cytoplasmic ALDOB staining (HPA tissue IHC). This paraffin-section guide covers the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet A03893-2) and chromogenic detection controls (standard IHC practice).
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 | Cytoplasmic staining in selected tissues (HPA tissue IHC) | |
| Staining pattern | Tubule cells, hepatocytes and intestinal endocrine cells: cytoplasmic (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03893-2) | |
| Caveat | Endogenous peroxidase may add DAB background in liver (standard IHC practice) | |
| Regulation | Expression regulation is unreported in the supplied evidence (UniProt; HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms; epitope location is unspecified (UniProt) |
Section 1
Recommended ALDOB IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published ALDOB staining methods for hepatocellular carcinoma, colorectal lesions, and gastric tissue (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 pH 6 retrieval (datasheet A03893-2); a gastric tissue protocol also reports high-pressure retrieval in 10 mM citrate at pH 6 (PMC5065259).
Section 2
What Is the Expected ALDOB Staining Pattern?
ALDOB should appear predominantly in the cytoplasm of renal tubular cells and hepatocytes, with staining also reported in intestinal endocrine cells (HPA tissue IHC: selective cytoplasmic expression; cell-level scores). This fits a cytosolic protein with no transmembrane segment (UniProt P05062: localization and topology). HPA rates the tissue IHC pattern Enhanced because antibody staining is highly consistent with RNA expression (HPA tissue IHC: reliability description).
| Strong cytoplasmic staining in kidney tubules and hepatocytes. | This matches the reported high cell-level signal in both tissues (HPA tissue IHC: kidney and liver, High). Judge the pattern in the named cells, alongside tissue morphology and the negative control (general IHC practice). |
| Cytoplasmic staining in intestinal endocrine cells, stronger in small intestine than duodenum. | The supplied cell-level scores are High in small intestine and Medium in duodenum (HPA tissue IHC). The broader tissue profile names both sites; it does not establish that every intestinal cell should stain (HPA tissue IHC: profile and cell-level scores). |
| Predominantly nuclear or crisp cell-surface staining in a known-positive tissue. | Those patterns disagree with the expected cytoplasmic distribution and lack of a transmembrane segment (HPA tissue IHC: profile; UniProt P05062: topology). Treat them as suspect and check morphology, detection controls and staining conditions before assigning ALDOB positivity (general IHC practice). |
| Strong staining in adipocytes or other cells scored Not detected. | Adipocytes in adipose tissue are scored Not detected (HPA tissue IHC). Consider nonspecific binding, cross-reactivity or endogenous detection activity; the image alone cannot distinguish these causes (general IHC practice). Compare an appropriate negative tissue and detection-only control (general IHC practice). |
| Weak or absent staining in otherwise interpretable kidney tubules or hepatocytes. | Both are reported High and serve as useful positive tissue comparators (HPA tissue IHC). A missing signal can reflect a staining-run problem or the sampled section; confirm tissue identity and controls, then review the validated assay conditions (general IHC practice). |
💡Expected ALDOB appearanceCall positive when renal tubular cells or hepatocytes show clear, predominantly cytoplasmic chromogenic signal consistent with their High HPA scores; isolated nuclear, cell-surface or broadly diffuse staining is suspect (HPA tissue IHC: cell scores and profile; UniProt P05062: topology; general IHC practice).
| Which compartment should guide scoring? | Cytosol is the primary expectation; UniProt also lists centrosome and centriolar satellite associations (UniProt P05062: localization). HPA tissue IHC reports a cytoplasmic pattern, so do not require visible centrosomal puncta to call a section positive (HPA tissue IHC: profile). |
| Which tissues provide the clearest comparison? | Kidney tubules and hepatocytes are High, intestinal endocrine cells are High in small intestine and Medium in duodenum, and adipocytes in adipose tissue are Not detected (HPA tissue IHC: cell-level scores). These observations support a tissue comparison, not a universal intensity cutoff. |
| How much confidence does antibody validation add? | Three listed antibodies have Enhanced IHC status (HPA antibodies: HPA002198, HPA073201, CAB020827). The tissue profile also has Enhanced reliability through agreement with RNA expression (HPA tissue IHC). Neither label establishes the behavior of an untested antibody or detection system. |
| IF/ICC Q&A: can this IHC pattern be assumed in cultured cells? | No cell lines with ICC-IF images or main subcellular location are supplied (HPA subcellular: unavailable). Cytosolic localization is reported by UniProt (UniProt P05062), but this payload cannot verify an IF/ICC staining pattern, intensity or cell-line control. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney or liver positive control has no chromogenic signal. | The expected High cell-level staining is missing (HPA tissue IHC: kidney tubules and hepatocytes). A failed staining or detection step is possible (general IHC practice). | Verify tissue morphology and control performance, then review the IHC-validated antibody's documented retrieval, dilution and detection conditions (general IHC practice). |
| Slide shows widespread haze that obscures cellular boundaries. | Diffuse background prevents reliable comparison with the selective cytoplasmic profile (HPA tissue IHC: profile). Nonspecific binding or detection background is possible (general IHC practice). | Compare the negative and detection-only controls; review blocking, washes and detection exposure, then rescore only interpretable cells (general IHC practice). |
| Signal is mainly nuclear or outlines cell membranes. | The compartment conflicts with cytoplasmic tissue staining and the absence of a transmembrane segment (HPA tissue IHC: profile; UniProt P05062: topology). | Check counterstain and tissue morphology, then compare positive and negative tissues with the same detection run (general IHC practice). |
| Adipocytes stain as strongly as the positive tissue. | Adipose-tissue adipocytes are scored Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity may explain the result, but staining alone cannot identify which (general IHC practice). | Inspect a detection-only control and compare the pattern against kidney tubules or hepatocytes stained in the same run (general IHC practice; HPA tissue IHC: High scores). |
| Duodenal staining appears weaker than small-intestine staining. | The supplied endocrine-cell scores differ: Medium in duodenum and High in small intestine (HPA tissue IHC). | Score the identified endocrine-cell compartment and tissue morphology; do not require equal intensity across the two sites (HPA tissue IHC: cell-level scores; general IHC practice). |
| A proposed IF/ICC control gives an ambiguous pattern. | HPA supplies no ICC-IF image-bearing cell lines or main subcellular assignment for ALDOB (HPA subcellular: unavailable). | Keep IF/ICC interpretation provisional and assess its own controls and antibody validation separately; the supplied IHC evidence cannot establish an IF/ICC result (HPA antibodies: ICC status unavailable; HPA subcellular). |
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 stain strongly (HPA: High in hepatocytes). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); on the liver slide, use non-hepatocyte areas to assess background without assuming those cells are ALDOB-negative (HPA: High in hepatocytes).
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 no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, plus ALDOB-knockout tissue as a biological negative (selected-SKU caption: rabbit primary antibody; standard IHC practice). Check for endogenous peroxidase and biotin background in liver because the reported detection uses an avidin-biotin complex and DAB (selected-SKU caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required (selected-SKU caption: citrate retrieval); the supplied evidence does not establish that frozen sections or IF are easier (HPA subcellular: no ICC-IF images). Liver endogenous biotin may add background with the reported avidin-biotin detection (selected-SKU caption: SABC; 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
Use the catalog antibody’s paraffin section result as the IHC starting point, then judge staining against ALDOB’s expected cell types and cytoplasmic localisation.
How should I troubleshoot weak ALDOB staining after antigen retrieval?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A03893-2). The selected paraffin section result used this treatment before overnight incubation with 1 μg/mL primary antibody at 4°C (selected IHC caption A03893-2). If staining is weak, compare retrieved and unretrieved serial sections, then adjust heating time in small steps while keeping antibody concentration and detection constant (standard IHC practice). Evaluate hepatocytes or renal tubule cells as expected positive populations (HPA tissue IHC). Excessive heating can damage morphology or increase diffuse staining, so judge signal alongside intact cell boundaries and a negative control (standard IHC practice).
Could fixation explain weak or uneven ALDOB staining?
The selected image documents a paraffin section but does not report its fixative, so ALDOB sensitivity to a particular fixation method or duration is unknown (selected IHC caption A03893-2). Record the fixative, time in fixative, and processing history for each specimen before comparing staining intensities (standard IHC practice). Compare serial sections from similarly processed material using the same citrate pH 6 retrieval and 20-minute heating step (datasheet A03893-2). If processing differs, treat an intensity difference as provisional until matched controls reproduce it (standard IHC practice). Preserve morphology when adjusting retrieval, because damaged tissue can make apparent cytoplasmic signal difficult to score (standard IHC practice).
What ALDOB staining pattern should I expect in tissue sections?
Expect predominantly cytoplasmic staining rather than a continuous membrane rim: ALDOB is annotated in the cytosol and has no transmembrane segment (UniProt P05062 localisation and topology). UniProt also annotates centrosome and centriolar satellite association, but the supplied tissue IHC profile describes selective cytoplasmic expression (UniProt P05062 localisation; HPA tissue IHC). Hepatocytes and renal tubule cells provide useful tissue landmarks with high reported staining (HPA tissue IHC). Check cell identity against the counterstain and section morphology before assigning punctate or perinuclear signal to ALDOB (standard IHC practice). Predominantly nuclear staining warrants review of antibody controls, retrieval, and chromogen deposition before biological interpretation (UniProt P05062 localisation; standard IHC practice).
How can I assess whether staining reflects ALDOB rather than epitope artefact?
UniProt lists 0 annotated ALDOB isoforms, but the supplied evidence does not map this antibody’s epitope or establish cross-reactivity with other aldolases (UniProt P05062; selected IHC caption A03893-2). Ask whether the antibody documentation identifies its immunogen before interpreting unexpected staining as an ALDOB variant (standard IHC practice). ALDOB has no annotated transmembrane segment and is primarily cytoplasmic, which provides a localisation check for the stain (UniProt P05062 topology and localisation). Compare expected positive cell populations with a no-primary control on adjacent sections (HPA tissue IHC; standard IHC practice). If specificity remains uncertain, concordance with an independently validated ALDOB antibody can strengthen the assignment (standard IHC practice).
How should I adapt these ALDOB localisation checks for multiplex IF?
For a companion IF experiment, pair ALDOB with a marker that identifies the expected cell population, such as hepatocytes or renal tubule cells, and assess overlap within individual cells (HPA tissue IHC; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, and reserve a spectrally distinct channel for the cell marker (standard IF practice). ALDOB is cytosolic and has no transmembrane segment, so use controlled permeabilisation to expose an intracellular epitope and check that morphology remains intact (UniProt P05062 localisation and topology; standard IF practice). Include single-stain and no-primary controls to identify bleed-through and background (standard IF practice). The supplied IHC caption does not establish IF performance for this antibody (selected IHC caption A03893-2).
How do I reduce diffuse or misleading chromogenic background?
The selected paraffin section workflow used 10% goat serum blocking, a biotinylated secondary, and DAB development (selected IHC caption A03893-2). Use a no-primary section to reveal secondary reagent or detection background, and include a peroxidase block in a DAB workflow (standard IHC practice). If diffuse colour persists, check washing, primary concentration, and chromogen development time while changing one variable at a time (standard IHC practice). A biotin-based detection system also merits an avidin–biotin background control where endogenous biotin is a concern (selected IHC caption A03893-2; standard IHC practice). Judge improvements against the expected cytoplasmic staining of hepatocytes or renal tubule cells (HPA tissue IHC).
How should I quantify ALDOB staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; hepatocytes and renal tubule cells are documented ALDOB-positive populations (HPA tissue IHC). For chromogenic sections, report an H-score from intensity and percentage of positive cells, or report percentage positive using a prespecified threshold (standard IHC practice). If counting discrete positive cells, express density per mm² of viable, analysable tissue rather than total slide area (standard IHC practice). Keep retrieval, DAB development, imaging, and threshold settings consistent across comparison groups (standard IHC practice). Normalise to the number or area of the relevant viable cells, and exclude folds, necrosis, and edge artefacts before comparing specimens (standard IHC practice).
When is apparent ALDOB positivity likely to be an artefact?
A credible result places staining mainly in cytoplasm and in cell populations consistent with the tissue context, including hepatocytes or renal tubule cells (UniProt P05062 localisation; HPA tissue IHC). Strong nuclear or membrane-rim signal conflicts with the supplied localisation and topology annotations and needs verification (UniProt P05062 localisation and topology). Inspect section edges, folds, and necrotic areas for concentrated DAB before scoring them as positive (standard IHC practice). Compare no-primary and peroxidase-block controls to identify detection background or endogenous peroxidase activity (standard IHC practice). Treat staining in an unexpected cell population as provisional until morphology, controls, and an independent specificity check support it (standard IHC practice).
Boster reagents
Best ALDOB / Fructose-bisphosphate aldolase B IHC Antibodies
A03893-2 has IHC images from human liver cancer and rat liver paraffin sections, plus IF/ICC images from A431, CACO-2 and U20S cells (catalog image captions).
Real IHC data
Anti-Aldolase/ALDOB Antibody ®
Cat # A03893-2
A03893-2 is listed for IHC and IF/ICC, with images from human liver cancer and rat liver paraffin sections (catalog applications; IHC image captions). Its IF/ICC images show A431, CACO-2 and U20S cells; listed reactivity covers human, monkey, mouse and rat (IF image captions; catalog reactivity).
Which to pick: Choose A03893-2 for paraffin-section IHC: its own captions document citrate retrieval at pH 6 for 20 minutes and primary antibody at 1 μg/ml; the fixative is unreported (A03893-2 IHC image captions). The same SKU is listed for IF/ICC at 2 μg/ml, with cell images in the catalog (catalog applications and dilution; A03893-2 IF image captions). For work across species, A03893-2 lists human, monkey, mouse and rat reactivity, though its IHC images document human and rat specimens; its host is rabbit and clonality is unreported (catalog reactivity and host; A03893-2 IHC image captions; catalog clone field).
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 Cytoplasmic staining in selected tissues (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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03893-2); Caveat: Endogenous peroxidase may add DAB background in liver (standard IHC practice); Isoform / epitope: No annotated isoforms; epitope location is unspecified (UniProt); Regulation: Expression regulation is unreported in the supplied evidence (UniProt; HPA tissue IHC). 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.