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ALDOC / Fructose-bisphosphate aldolase C · IHC design guide
Design Immunohistochemistry for ALDOC
Plan ALDOC staining in paraffin sections around the selective cytoplasmic signal seen in subsets of neurons and glial cells (HPA tissue IHC). Use the catalog antibody’s IHC conditions and compare stained cells with nearby unstained cells.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ALDOC 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 in subsets of neurons and glia (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic staining in neurons and glia (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05296-1) | |
| Positive control | Caudate+4 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 A05296-1) | |
| Caveat | Only subsets of brain cells stain; nearby cells may be negative (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; no processing-based epitope shift (UniProt) |
Section 1
Recommended ALDOC IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A05296-1) is accompanied by published ALDOC staining protocols for colorectal and gastric cancer tissue (PMC12168302; PMC11565221).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A05296-1) |
| Fixation | Image fixative and duration unreported (datasheet A05296-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A05296-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05296-1) |
| Primary antibody | Rabbit anti-ALDOC, 2-5 μg/ml (datasheet A05296-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05296-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05296-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ALDOC-positive staining in glial cells of caudate (HPA tissue IHC: Medium). HPA tissue profile: Selective cytoplasmic expression in subsets of neurons and glial cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A05296-1). Use the reported citrate retrieval when reproducing either published protocol (PMC12168302; PMC11565221).
Section 2
What Is the Expected ALDOC Staining Pattern?
For paraffin-section IHC, expect selective cytoplasmic ALDOC staining in subsets of neurons and glial cells (HPA tissue IHC: Enhanced reliability). Glial cells in caudate, cerebral cortex, and hippocampus are documented positives, as are processes in cerebellar white matter (HPA tissue IHC: Medium). ALDOC has no annotated transmembrane segment (UniProt P09972 topology). HPA ICC-IF reports additional locations, addressed separately below.
| Selective cytoplasmic stain in some glial cells, with unstained neighbors (HPA tissue IHC: selective expression). | This fits the reported brain IHC pattern. Judge the labeled cells and their cytoplasm, rather than expecting every cell in the section to stain (HPA tissue IHC: selective cytoplasmic expression). |
| Predominantly nuclear stain in paraffin brain sections, without the expected cytoplasmic pattern. | Question the IHC result against HPA's tissue pattern (HPA tissue IHC: cytoplasmic). Nuclear ALDOC is reported in ICC-IF, so compartment alone cannot prove artefact across methods (HPA subcellular ICC-IF: nucleoplasm and nucleoli fibrillar center). |
| Strong, widespread stain in an HPA-negative reference cell population, such as adipocytes. | Suspect nonspecific binding, cross-reactivity, or endogenous chromogenic activity; check controls before assigning ALDOC expression (standard IHC practice). HPA reports adipocytes in adipose tissue as not detected (HPA tissue IHC). |
| Diffuse color over tissue and spaces between cells obscures cell boundaries. | Treat this as background until a cell-restricted pattern is resolved (standard IHC practice). HPA's reported ALDOC signal is selective within cells and processes, not an all-over deposit (HPA tissue IHC). |
| No cytoplasmic signal in caudate glial cells or cerebellar white-matter processes. | A known-positive reference has failed; review the run before interpreting other sections as ALDOC-negative (HPA tissue IHC: Medium in both populations; standard IHC practice). |
💡Expected ALDOC appearanceCall a result consistent with HPA when Medium, selective cytoplasmic staining marks glial cells or cerebellar white-matter processes; uniform staining of HPA-negative cell populations warrants a false-positive check (HPA tissue IHC; standard IHC practice).
| Tissue and cell selection | Brain and retina are RNA group enriched, while the supplied tissue IHC examples document Medium signal in selected brain cells and late or elongated spermatids (HPA RNA specificity; HPA tissue IHC). Do not infer that every cell in those tissues is positive. |
| Cellular compartment and topology | HPA describes selective cytoplasmic tissue IHC, and UniProt annotates no transmembrane segment (HPA tissue IHC; UniProt P09972 topology). A membrane-rim pattern alone would need independent validation (standard IHC practice). |
| Antibody evidence | HPA lists IHC as Enhanced for HPA003282 and CAB020828 and Supported for HPA067442 (HPA antibodies). These grades support checking the expected pattern, but do not establish performance for an unspecified catalog antibody. |
| Protein form and epitope limits | UniProt lists one chain spanning residues 1–364, with no signal peptide, propeptide, or annotated isoforms; it also lists modified residues (UniProt P09972). No antibody epitope or ALDOC-specific retrieval response is supplied, so neither predicts a staining shift. |
| IF/ICC Q: Should nuclear staining be expected? | A: In ICC-IF, HPA reports approved nucleoplasm and nucleoli fibrillar-center localization, plus vesicles and cytosol (HPA subcellular ICC-IF). Use the cytoplasmic tissue-IHC profile when judging paraffin-section chromogenic IHC (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive brain control has no visible signal. | The run may have failed at retrieval, primary-antibody binding, or detection; the image alone cannot identify which step (standard IHC practice). | Confirm tissue identity and include the documented positive cell population; then review retrieval, primary incubation, and detection controls using the antibody's IHC-P instructions (HPA tissue IHC: Medium brain examples; standard IHC practice). |
| Most cells stain equally in a section expected to show selective labeling. | Excess primary concentration or nonspecific detection may obscure selectivity (standard IHC practice). | Compare with a no-primary control and titrate the IHC-validated antibody; check whether HPA-negative reference cells remain unstained (standard IHC practice; HPA tissue IHC: adipocytes not detected). |
| Brown signal appears in negative-reference cells despite no primary antibody. | Endogenous detection activity or substrate deposition can generate chromogen without specific ALDOC binding (standard IHC practice). | Review endogenous-enzyme blocking and detection reagents; interpret the negative reference at the cell-population level (standard IHC practice; HPA tissue IHC: adipocytes not detected). |
| Only nuclei stain in paraffin brain IHC. | This differs from the supplied tissue-IHC profile, although HPA separately reports nuclear ICC-IF localization (HPA tissue IHC; HPA subcellular ICC-IF). | Check the no-primary control and compare cytoplasmic staining in documented positive cells before calling the IHC pattern specific (standard IHC practice; HPA tissue IHC: glial cells Medium). |
| A cerebellar section seems negative when only cell bodies are inspected. | The supplied positive example identifies processes in white matter, which can be missed by cell-body-only scoring (HPA tissue IHC: cerebellum, Medium). | Inspect white-matter processes and assess whether their signal is selective and above the run's background (HPA tissue IHC; standard IHC practice). |
| Color is diffuse or counterstain makes the positive cells hard to distinguish. | Background or an overly strong counterstain can make cellular localization difficult to score (standard IHC practice). | Use the no-primary control to gauge background, review washes and detection development, and adjust counterstaining so cytoplasmic signal can be judged (standard IHC practice; HPA tissue IHC: selective cytoplasmic pattern). |
Experiment evidence
IHC & IF images showing ALDOC staining
A05296-1 · IHCIHC analysis of Aldolase C/ALDOC using anti-Aldolase C/ALDOC antibody (A05296-1). Aldolase C/ALDOC was detected in a paraffin-embedded secti
A05296-1 · IHCIHC analysis of Aldolase C/ALDOC using anti-Aldolase C/ALDOC antibody (A05296-1). Aldolase C/ALDOC was detected in a paraffin-embedded secti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109107-ALDOC/tissue/cerebral+cortex#imid_5849016
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000109107-ALDOC/tissue/cerebellum#imid_5849027
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109107-ALDOC/tissue/colon#imid_5848958
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109107-ALDOC/tissue/liver#imid_5848943
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003282; green = ALDOC, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003282; green = ALDOC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ALDOC IHC & IF
🧪Run caudate first and expect staining in glial cells (HPA: Caudate glial cells, Medium). Use adipose tissue as a negative comparator and assess its adipocytes (HPA: Adipose tissue adipocytes, Not detected); on the caudate slide, cells without specific staining should remain at background, although the supplied HPA rows do not identify a named internal negative cell type.
Positive control tissue: Caudate (Glial cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ALDOC in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched nonimmune rabbit IgG control for the catalog antibody (selected-SKU caption: rabbit antibody), and an ALDOC knockout specimen as a biological negative. For chromogenic detection, quench endogenous peroxidase before applying the peroxidase-conjugated secondary (selected-SKU caption: peroxidase secondary and DAB); assess tissue autofluorescence separately if using IF.
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the exact selected-SKU paraffin-section caption also leaves the fixative unreported (selected-SKU caption: fixative not stated). That caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU caption: EDTA retrieval in a paraffin section). For brain IF, check tissue autofluorescence before interpreting signal; the supplied evidence does not report a tissue-specific artefact.
HPA tissue IHC evidence for ALDOC
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 |
|---|---|---|---|---|
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Processes in white matter | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | 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 ALDOC IHC Tips
Troubleshoot ALDOC staining in paraffin sections by checking retrieval, cell type, compartment and controls against the reported tissue pattern (HPA tissue IHC; HPA subcellular).
What retrieval should I use when ALDOC staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 for ALDOC paraffin sections (datasheet A05296-1). The reported rat brain staining used this retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A05296-1). If staining remains weak, compare a citrate buffer at pH 6.0 on adjacent sections while holding detection and development conditions constant (standard IHC practice). Include a known positive brain section and a no-primary control in each comparison (HPA tissue IHC; standard IHC practice). Score signal in expected cells alongside tissue damage and background, since stronger colour alone does not establish improved specificity (HPA tissue IHC; standard IHC practice).
Could fixation explain variable ALDOC staining between paraffin specimens?
The ALDOC paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A05296-1). Record each specimen's fixative, fixation duration and processing history before comparing staining intensity across specimens (standard IHC practice). Run similarly processed positive brain tissue with each batch, and keep EDTA retrieval at pH 8.0 and antibody concentration at 2 μg/ml constant during the comparison (HPA tissue IHC; datasheet A05296-1). Inspect morphology and staining distribution together, because damaged or poorly preserved tissue can complicate interpretation of chromogenic signal (standard IHC practice). Do not infer an ALDOC-specific fixation effect from its reported tissue pattern or protein annotations (HPA tissue IHC; UniProt P09972).
How should I assess cytoplasmic and nuclear ALDOC staining in tissue?
Expect selective cytoplasmic staining in subsets of neurons and glial cells when assessing tissue IHC (HPA tissue IHC). Reported positive sites include glial cells in caudate, cerebral cortex and hippocampus, plus processes in cerebellar white matter (HPA tissue IHC). Cell-based IF observations also place ALDOC in nucleoplasm and nucleoli, with additional cytosolic and vesicular signal; these observations warrant checking nuclear staining rather than automatically dismissing it (HPA subcellular). UniProt supplies no subcellular localisation annotation for ALDOC, so annotate the compartment and cell type seen in each section (UniProt P09972; standard IHC practice). Compare unusual patterns with a positive control and no-primary section before assigning them to ALDOC (standard IHC practice).
How can I investigate unexpected ALDOC staining across cell types?
The supplied ALDOC record lists 0 isoforms and one chain spanning residues 1–364; an isoform switch therefore has no support here (UniProt P09972). ALDOC has no annotated transmembrane segment or signal peptide, so sharply membranous staining merits scrutiny against the expected cytoplasmic tissue pattern (UniProt P09972 topology; HPA tissue IHC). Its annotated modifications include phosphorylation and lysine acetylation, but the supplied evidence does not identify this antibody's epitope or show an effect of those modifications on staining (UniProt P09972; datasheet A05296-1). Check antibody documentation for epitope information, then compare tissue distribution with appropriate positive and negative controls (standard IHC practice). Avoid assigning a staining difference to a particular modification without direct evidence (standard IHC practice).
How should I adapt ALDOC localisation checks to multiplex IF?
Pair ALDOC with a validated glial marker when testing the reported glial staining pattern, and assess overlap at the level of individual cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, favouring a far-red channel for weak ALDOC signal when tissue autofluorescence is prominent, and include single-stain controls (standard IF practice). ALDOC has no annotated transmembrane segment; use a controlled permeabilisation step when testing access to cytosolic or nuclear epitopes, while recognising that this antibody's epitope is unspecified (UniProt P09972 topology; standard IF practice). Cell-based IF reports nucleoplasmic, nucleolar, vesicular and cytosolic localisation, so document compartment separately from cell identity (HPA subcellular). Compare signal with no-primary and autofluorescence controls before interpreting apparent colocalisation (standard IF practice).
What should I check when DAB obscures ALDOC-positive cells?
The reported ALDOC paraffin-section workflow used 10% goat serum blocking, a peroxidase-conjugated secondary antibody and DAB development (datasheet A05296-1). Add an endogenous peroxidase block as a general chromogenic IHC step, and examine a no-primary section for residual enzyme or secondary-antibody background (standard IHC practice). Compare background in expected negative cells with staining in glial cells of a positive brain section before adjusting conditions (HPA tissue IHC; standard IHC practice). Keep retrieval and development conditions matched across comparison sections so a change in colour can be attributed more clearly (standard IHC practice). If diffuse colour persists, reassess blocking, washes and development time using the controls (standard IHC practice).
How should I quantify ALDOC staining across brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and score relevant cells separately, because tissue IHC reports selective staining in subsets of neurons and glial cells (HPA tissue IHC; standard IHC practice). For cellular staining, record the percentage of positive cells and an intensity-based H-score; document the positivity threshold before reviewing groups (standard IHC practice). For stained processes, measure signal-positive area or density per mm² within a consistently defined region instead of forcing a cell-level score (HPA tissue IHC; standard IHC practice). Normalise cellular counts to the number of eligible cells and area measures to analysed tissue area, excluding folds and missing tissue by the same rules (standard IHC practice). Process comparison sections together and report compartment-specific results (standard IHC practice).
When is an ALDOC IHC signal likely to be artefactual?
Give greatest weight to selective staining in the reported neuronal or glial subsets and cerebellar white-matter processes, with suitable positive and no-primary controls (HPA tissue IHC; standard IHC practice). Predominantly sharp membranous staining warrants review because ALDOC has no annotated transmembrane segment and tissue IHC reports a cytoplasmic pattern (UniProt P09972 topology; HPA tissue IHC). Nuclear signal alone is inconclusive: cell-based IF reports nucleoplasmic and nucleolar ALDOC, while the reported tissue IHC pattern is cytoplasmic (HPA subcellular; HPA tissue IHC). Review section edges, necrotic regions and no-primary staining for processing artefact or endogenous peroxidase before calling diffuse DAB colour positive (standard IHC practice). Record discrepant cell types and compartments explicitly (standard IHC practice).
Boster reagents
Best ALDOC / Fructose-bisphosphate aldolase C IHC Antibodies
A05296-1 has real IHC images from paraffin-embedded rat and human brain sections; listed reactivity includes human, mouse, and rat (A05296-1 image captions; catalog: reactivity).
Real IHC data
Anti-Aldolase C/ALDOC Antibody ®
Cat # A05296-1
A05296-1 is the only rendering SKU and is listed for IHC and WB (catalog: applications). Its IHC captions show paraffin-embedded rat and human brain sections with EDTA pH 8.0 retrieval and 2 μg/ml primary antibody (A05296-1 image captions).
Which to pick: For tissue IHC, choose A05296-1 for paraffin sections using its documented retrieval and staining conditions (A05296-1 image captions). No IF/ICC choice is supported: A05296-1 has no listed IF application or IF image (catalog: applications; IF images). For cross-species planning, A05296-1 is rabbit polyclonal with listed human, mouse, and rat reactivity, while its IHC images cover rat and human brain only; the fixative is unreported (catalog: host, clonality, reactivity; A05296-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P09972 (ALDOC_HUMAN, Fructose-bisphosphate aldolase C).
- Human Protein Atlas. ALDOC tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ALDOC subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli fibrillar center. In addition localized to vesicles and cytosol..
- Human Protein Atlas. ALDOC antibody validation summary (3 antibodies).
- Enrichment of Aldolase C Correlates with Low Non-Mutated IDH1 Expression and Predicts a Favorable Prognosis in Glioblastomas. Cancers 2019 — PMC6770576.
- ALDOC and PGK1 coordinately induce glucose metabolism reprogramming and promote development of colorectal cancer. Molecular medicine (Cambridge, Mass.) 2025 — PMC12168302.
- Fructose-bisphosphate Aldolase C Expression is Associated with Poor Prognosis and Stemness in Gastric Cancer. Acta histochemica et cytochemica 2024 — PMC11565221.
- Region-Restrict Astrocytes Exhibit Heterogeneous Susceptibility to Neuronal Reprogramming. Stem cell reports 2019 — PMC6373495.
- PubMed PMID:3105602 — UniProt-cited evidence.
- PubMed PMID:3267224 — UniProt-cited evidence.
- PubMed PMID:2209624 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ALDOC (Fructose-bisphosphate aldolase C). Expected localisation Cytoplasmic in subsets of neurons and glia (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05296-1). Validated IHC antibody: A05296-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A05296-1); 2. Fixation: Image fixative and duration unreported (datasheet A05296-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A05296-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05296-1); 8. Primary antibody: Rabbit anti-ALDOC, 2-5 μg/ml (datasheet A05296-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05296-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A05296-1); Caveat: Only subsets of brain cells stain; nearby cells may be negative (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; no processing-based epitope shift (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt P09972, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm and nucleoli fibrillar center.…, Datasheet A05296-1, PMC12168302, PMC11565221. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ALDOC/immunohistochemistry-ihc-guide.