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ALDOA / Fructose-bisphosphate aldolase A · IHC design guide
Design Immunohistochemistry for ALDOA
Plan chromogenic ALDOA IHC in paraffin sections using the observed cytoplasmic tissue pattern (HPA tissue IHC). Use skeletal myocytes as a high-staining reference and score signal by cell type and compartment (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ALDOA 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 most cell types (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic staining, including skeletal myocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A05022-3) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Caudate+1 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). Selected-image fixative and duration unreported (datasheet A05022-3); verify before use. | |
| Caveat | Skeletal myocytes stain highly; cardiomyocytes are undetected (HPA tissue IHC) | |
| Regulation | Cell-type-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended ALDOA IHC & IF Protocols
The catalog antibody uses citrate pH 6 retrieval (datasheet A05022-3). One published LUSC IHC protocol uses microwave EDTA pH 9.0 (PMC12619220: IHC methods).
| Sample | Paraffin-embedded human Lung cancer tissues; fixative not specified (datasheet A05022-3) |
| Fixation | Image fixative and duration unreported (datasheet A05022-3); 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 A05022-3) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05022-3) |
| Primary antibody | Rabbit anti-ALDOA, 0.5-1μg/ml (datasheet A05022-3) |
| Primary incubation | Overnight at 4 °C (datasheet A05022-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05022-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ALDOA-positive staining in purkinje cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression at variable levels in most cell types, including skeletal myocytes. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 for the catalog antibody (datasheet A05022-3); consider EDTA pH 9.0 if staining is weak (PMC12619220: IHC methods).
Section 2
What Is the Expected ALDOA Staining Pattern?
ALDOA should stain mainly in the cytoplasm, including skeletal myocytes; Purkinje cells also show cytoplasmic and membrane staining (HPA: tissue IHC). In skeletal muscle, ALDOA associates with sarcomeric I bands and M lines (UniProt P04075: subcellular location). It has no transmembrane segment (UniProt P04075: topology). Treat tissue intensity as a guide rather than an absolute rule: HPA rates its IHC profile Supported, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Myocytes stain strongly in their cytoplasm; neighboring cells may differ in intensity. | This matches high myocyte staining in skeletal muscle and variable cytoplasmic expression across most cell types (HPA: tissue IHC). Assess the cell containing the signal, not just the overall darkness of the section (standard IHC practice). |
| Signal is predominantly nuclear, or appears only as a sharp cell-edge outline outside the reported Purkinje-cell pattern. | An isolated nuclear or cell-edge pattern is inconsistent with ALDOA’s reported cytosolic location (HPA: subcellular ICC-IF; UniProt P04075: subcellular location). Check morphology and detection controls before calling it specific; HPA does report cytoplasm/membrane staining in Purkinje cells (HPA: tissue IHC). |
| An unexpected cell population stains more strongly than the expected cells in the same section. | Compare its identity and compartment with HPA’s cell-level observations before interpreting it as ALDOA. Broad cytoplasmic expression is reported, so unexpected staining alone does not prove cross-reactivity; staining persisting without primary antibody can indicate endogenous detection activity (HPA: tissue IHC; standard IHC practice). |
| Brown reaction product is diffuse across tissue, extracellular spaces, or the slide. | A uniform haze that obscures cell boundaries cannot establish ALDOA localisation. In chromogenic IHC, incomplete blocking, excess antibody or detection reagent, and inadequate washing are general causes to investigate (standard IHC practice). |
| A skeletal-muscle section has no myocyte signal. | HPA reports high myocyte staining there, making it a useful positive comparator, but its Supported rating and medium RNA agreement do not make every section guaranteed positive (HPA: tissue IHC). First verify tissue preservation and the assay’s positive control, then examine retrieval and detection steps (standard IHC practice). |
💡Expected ALDOA appearanceCall a result consistent with ALDOA when staining is predominantly cytoplasmic and strong in skeletal myocytes, with variable cytoplasmic staining in other cell types; isolated nuclear signal or diffuse cell-free deposit is suspect (HPA: tissue IHC and subcellular ICC-IF; standard IHC practice).
| Compartment and sarcomeric context | Cytosol is the approved ICC-IF location; UniProt also places ALDOA at skeletal-muscle I bands and M lines (HPA: subcellular ICC-IF; UniProt P04075: subcellular location). On routine chromogenic sections, judge cell-level cytoplasmic staining before attempting to resolve fine sarcomeric detail (standard IHC practice). |
| Tissue and cell-level contrast | Skeletal myocytes and Purkinje cells are High; adipocytes and several glandular or epithelial populations are Medium, while HPA lists cardiomyocytes and caudate glial cells as Not detected (HPA: tissue IHC). These are observed patterns, not universal positive or negative guarantees (HPA: Supported reliability, medium RNA agreement). |
| Intracellular topology | UniProt annotates no transmembrane segment or signal peptide (UniProt P04075: topology and processing). Together with the approved cytosolic ICC-IF location, this supports an intracellular expectation; it does not define how fixation or antigen retrieval affects a particular antibody (HPA: subcellular ICC-IF). |
| Isoforms and antibody interpretation | Two isoforms are listed, but the supplied record gives no isoform-specific tissue staining or antibody epitope (UniProt P04075: isoforms; HPA: antibody summary). Do not assign a staining difference to an isoform or choose retrieval conditions from isoform count alone. |
| Antibody evidence | HPA004177 and CAB006252 are each IHC Supported; HPA004177 is also ICC Approved (HPA: antibody summary). These validation labels support use of the reported pattern as a comparator, but do not establish that either antibody performs identically under every staining condition. |
| What should IF/ICC show? | Predominantly cytosolic ALDOA is the approved ICC-IF pattern, with images listed for A-431, U-251MG, U2OS, RPTEC/TERT1 and serum-starved hTERT-RPE1 (HPA: subcellular ICC-IF). Use that localisation as a cross-check; this IHC section does not specify an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Skeletal myocytes are weak or unstained. | The expected High myocyte signal is missing, but HPA does not identify a target-specific fixation or retrieval failure mechanism (HPA: tissue IHC). | Check tissue morphology and a known-positive section, then review retrieval, primary-antibody application and chromogenic detection against the validated IHC workflow (standard IHC practice). |
| Every cell appears equally dark. | This obscures HPA’s variable cell-level cytoplasmic profile and may reflect excessive reagent or inadequate washing (HPA: tissue IHC; standard IHC practice). | Compare with a section processed without primary antibody; review antibody concentration, blocking, wash steps and development time (standard IHC practice). |
| A no-primary control develops brown signal. | The signal does not require primary-antibody binding; endogenous enzyme activity or nonspecific detection-system binding is possible (standard IHC practice). | Apply the appropriate endogenous-enzyme block for the detection chemistry and check secondary or polymer controls (standard IHC practice). |
| Signal is mainly nuclear. | That location conflicts with approved cytosolic ICC-IF staining and UniProt’s cytoplasmic annotation (HPA: subcellular ICC-IF; UniProt P04075: subcellular location). | Inspect the counterstain and cell boundaries, compare a no-primary control, and reassess antibody concentration and detection conditions (standard IHC practice). |
| Cardiomyocytes stain strongly while the expected positive comparator is weak. | HPA lists cardiomyocytes as Not detected and skeletal myocytes as High; the reversal warrants investigation but is not, by itself, proof of antibody cross-reactivity (HPA: tissue IHC). | Confirm tissue and cell identity, inspect the compartment, and compare a known-positive section and no-primary control in the same run (standard IHC practice). |
| One antibody yields a different pattern from another. | Both listed antibodies are IHC Supported, but the supplied evidence does not establish identical epitopes or identical behavior in every preparation (HPA: antibody summary). | Compare each result with cell-level HPA observations and its own controls; document retrieval and detection conditions before attributing the difference to ALDOA biology (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing ALDOA staining
A05022-3 · IHCIHC analysis of ALDOA using anti-ALDOA antibody (A05022-3). ALDOA was detected in paraffin-embedded section of human Lung cancer tissues. He
A05022-3 · IHCIHC analysis of ALDOA using anti-ALDOA antibody (A05022-3). ALDOA was detected in paraffin-embedded section of mouse heart tissues. Heat med
A05022-3 · IFIF analysis of Aldolase/ALDOA using anti- Aldolase/ALDOA antibody (A05022-3). Aldolase/ALDOA was detected in immunocytochemical section of A
M05022-2 · IHCIHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (M05022-2). Aldolase/ALDOA was detected in paraffin-embedded section of hu
M05022-2 · IHCIHC analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (M05022-2). Aldolase/ALDOA was detected in paraffin-embedded section of hu
M05022-2 · IFIF analysis of Aldolase/ALDOA using anti-Aldolase/ALDOA antibody (M05022-2). Aldolase/ALDOA was detected in immunocytochemical section of HE
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149925-ALDOA/tissue/cerebral+cortex#imid_1521448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149925-ALDOA/tissue/colon#imid_1521374
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149925-ALDOA/tissue/liver#imid_1521359
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000149925-ALDOA/tissue/kidney#imid_1521368
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004177; green = ALDOA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004177; green = ALDOA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ALDOA IHC & IF
🧪Run skeletal muscle first: myocytes must show strong ALDOA staining (HPA: High in myocytes), consistent with cytoplasmic and myofibrillar localisation (UniProt P04075 subcellular location). Use heart muscle as the negative tissue (HPA: Not detected in cardiomyocytes); on the skeletal muscle slide, compare interstitial non-myocytes with myocytes for background or weaker staining, without assuming those cells are ALDOA-negative (HPA: High in myocytes only).
Positive control tissue: Cerebellum (Purkinje cells - cytoplasm/membrane, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ALDOA in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (catalog antibody caption: rabbit primary), plus ALDOA-knockout tissue as a biological negative where available. Quench endogenous peroxidase and assess endogenous biotin background in skeletal muscle because the reported IHC detection uses a streptavidin–biotin complex with DAB (catalog antibody caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU A05022-3 tissue-IHC caption does not state a fixative (catalog antibody caption: fixative unreported). That paraffin-section example uses heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish whether retrieval is required for every specimen (catalog antibody caption). The supplied evidence does not show that frozen sections or IF/ICC are easier; for skeletal muscle, assess staining against its structured myofibrillar localisation when scoring (UniProt P04075 subcellular location).
HPA tissue IHC evidence for ALDOA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ALDOA IHC Tips
Troubleshoot ALDOA staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls, and cell-specific scoring before interpreting signal intensity.
What retrieval should I try when ALDOA staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A05022-3). The catalog antibody detected ALDOA in a paraffin section of human lung cancer tissue after that treatment, followed by 1 μg/mL primary antibody overnight at 4°C (datasheet A05022-3). If staining remains weak, check heating uniformity, section adhesion, antibody incubation, and detection before changing retrieval conditions (standard IHC practice). Run a known positive section alongside the specimen, and compare cytoplasmic signal in matched cell types rather than judging the whole section by its darkest area (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or patchy ALDOA staining?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A05022-3). Record the specimen’s fixative and fixation interval, then compare sections processed together using the same citrate pH 6, 20-minute retrieval and detection conditions (datasheet A05022-3; standard IHC practice). Uneven fixation or processing can produce uneven staining in paraffin sections, so inspect morphology and compare the centre with the edges before attributing a gradient to ALDOA (standard IHC practice). Include a concurrently processed positive section; its cytoplasmic staining helps distinguish a run-level problem from a specimen-specific one (HPA tissue IHC; standard IHC practice).
Which ALDOA staining pattern should I expect in paraffin sections?
Expect predominantly cytoplasmic staining across many cell types; HPA reports variable levels and rates its tissue IHC evidence as Supported, with medium RNA–protein consistency (HPA tissue IHC). In skeletal myocytes, ALDOA can also associate with sarcomeric I bands and M lines, although ordinary chromogenic sections may not resolve that fine pattern (UniProt P04075 subcellular location; standard IHC practice). HPA reports high staining in skeletal myocytes and Purkinje cells, while caudate glial cells and cardiomyocytes are listed as not detected (HPA tissue IHC). Judge a suspicious membrane-only or nuclear-only pattern against cell morphology and a positive control before treating it as ALDOA (HPA subcellular; standard IHC practice).
How should isoforms and epitope uncertainty affect ALDOA IHC optimisation?
ALDOA has 2 annotated isoforms, but the supplied evidence does not map the catalog antibody’s epitope or establish which isoforms it detects (UniProt P04075 isoforms; datasheet A05022-3). Its annotated modified residues include phosphorylation and acetylation sites, so epitope accessibility cannot be inferred from the protein name alone (UniProt P04075 modified residues; standard IHC practice). Keep citrate pH 6 retrieval at 20 minutes as the reference condition while comparing any optimisation on adjacent sections (datasheet A05022-3; standard IHC practice). If staining differs between samples, document processing and cell composition before assigning the difference to an isoform or modification (standard IHC practice).
How can I check an ALDOA pattern by multiplex IF?
Use a marker for the expected cell type, such as a myocyte marker in skeletal muscle, so ALDOA signal can be assigned to cells rather than nearby structures (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores and consider a far-red ALDOA channel when the tissue shows strong autofluorescence; inspect an unstained section in each channel (standard IF practice). ALDOA is cytosolic and has no annotated transmembrane segment, so permeabilisation should give the antibody access to a cytosolic epitope; the catalog epitope itself is unspecified (HPA subcellular; UniProt P04075 topology; datasheet A05022-3). Optimise IF fixation, permeabilisation, and antibody concentration independently of the paraffin-section IHC conditions, and include single-colour controls to assess bleed-through (standard IF practice).
What should I check when ALDOA DAB staining looks diffuse or excessive?
First compare the section with a no-primary control to identify signal from detection reagents or endogenous activity (standard chromogenic IHC practice). The catalog tissue example used biotinylated secondary antibody, a streptavidin–biotin complex, and DAB, so assess background from that detection sequence under matched conditions (datasheet A05022-3; standard IHC practice). Apply a peroxidase block and review serum blocking, washes, and DAB development as general workflow controls; the caption specifies 10% goat serum but gives no peroxidase-block condition (datasheet A05022-3; standard IHC practice). Because ALDOA is broadly cytoplasmic, diffuse colour alone is insufficient: look for cell-associated signal above the no-primary control (HPA tissue IHC; standard IHC practice).
How should I quantify heterogeneous ALDOA staining across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions before scoring, then record the percentage of positive cells and staining intensity to calculate an H-score where morphology permits (standard IHC practice). For each region, use H-score = 1×(% weak) + 2×(% moderate) + 3×(% strong), yielding a 0–300 scale (standard IHC practice). Alternatively, report positive-cell density per mm² with the counted tissue area, while excluding folds, necrosis, and unusable edges by predefined rules (standard IHC practice). Normalise comparisons to the same cell type, viable tissue area, staining run, and scoring thresholds, because ALDOA levels vary among cell types (HPA tissue IHC; standard IHC practice).
How do I distinguish true ALDOA signal from artefact?
A credible ALDOA signal is cell-associated and mainly cytoplasmic, consistent with its approved cytosolic location and variable cytoplasmic tissue profile (HPA subcellular; HPA tissue IHC). Compare expected positive populations, including skeletal myocytes, with the relevant morphology; HPA also lists cardiomyocytes as not detected, so muscle identity alone does not predict positivity (HPA tissue IHC). Treat isolated nuclear or membrane-only colour, strong section-edge gradients, and staining confined to necrosis as patterns requiring control review (HPA subcellular; standard IHC practice). Check the no-primary control and peroxidase block for endogenous enzyme or detection background before interpreting DAB colour as ALDOA (standard chromogenic IHC practice).
Boster reagents
Best ALDOA / Fructose-bisphosphate aldolase A IHC Antibodies
Anti-ALDOA antibodies have IHC images from human cancer tissues and mouse heart, plus IF images from A431 and HEPG2 cells (catalog image captions).
Real IHC data
Anti-Aldolase/ALDOA Antibody ®
Cat # A05022-3
Real IHC data
Anti-Aldolase/ALDOA Antibody ® (monoclonal, 6H8)
Cat # M05022-2
A05022-3 has paraffin-section IHC images from human lung, intestinal and mammary cancer tissues and mouse heart, plus IF data from A431 cells (catalog image captions). M05022-2 has paraffin-section IHC images from human liver, breast and rectal cancer tissues, plus IF data from HEPG2 cells (catalog image captions).
Which to pick: For human tissue IHC, choose A05022-3 with citrate retrieval at pH 6, or mouse monoclonal M05022-2 (clone 6H8) with EDTA retrieval at pH 8; each condition comes from that SKU’s own paraffin-section IHC caption, and neither caption reports the fixative (catalog IHC image captions; catalog clone). For IF/ICC, both have cell images: A05022-3 in A431 cells and M05022-2 in HEPG2 cells (catalog IF image captions). For human, mouse or rat samples, A05022-3 lists reactivity with all three and has a mouse-heart IHC image; M05022-2 lists human reactivity only (catalog reactivity; A05022-3 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P04075 (ALDOA_HUMAN, Fructose-bisphosphate aldolase A).
- Human Protein Atlas. ALDOA tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ALDOA subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. ALDOA antibody validation summary (2 antibodies).
- Knockout of the sulfide: quinone oxidoreductase SQR reduces growth of HCT116 tumor xenograft. Redox biology 2025 — PMC12433914.
- Aldolase A promotes epithelial-mesenchymal transition to increase malignant potentials of cervical adenocarcinoma. Cancer science 2020 — PMC7419050.
- Aldolase a in pan-cancer and lung squamous cell carcinoma: prognostic value and macrophage-driven immune suppression unveiled by multi-omics and cohort validation. Cancer cell international 2025 — PMC12619220.
- High expression of Aldolase A predicts poor survival in patients with clear-cell renal cell carcinoma. Therapeutics and clinical risk management 2017 — PMC5338975.
- PubMed PMID:3840020 — UniProt-cited evidence.
- PubMed PMID:3030757 — UniProt-cited evidence.
- PubMed PMID:3391172 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ALDOA (Fructose-bisphosphate aldolase A). Expected localisation Cytoplasmic staining in most cell types (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A05022-3). Validated IHC antibody: A05022-3 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells - cytoplasm/membrane); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Lung cancer tissues; fixative not specified (datasheet A05022-3); 2. Fixation: Image fixative and duration unreported (datasheet A05022-3); 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 A05022-3); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05022-3); 8. Primary antibody: Rabbit anti-ALDOA, 0.5-1μg/ml (datasheet A05022-3); 9. Primary incubation: Overnight at 4 °C (datasheet A05022-3). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A05022-3); verify before use.; Caveat: Skeletal myocytes stain highly; cardiomyocytes are undetected (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences are unspecified (UniProt); Regulation: Cell-type-dependent expression (HPA tissue IHC). Evidence: UniProt P04075, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A05022-3, PMC12619220. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ALDOA/immunohistochemistry-ihc-guide.