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AGPS / Alkyldihydroxyacetonephosphate synthase, peroxisomal · IHC design guide
Design Immunohistochemistry for AGPS
Plan AGPS chromogenic IHC in paraffin sections using the catalog antibody’s 2–5 μg/ml range (datasheet A03481-1). Compare granular cytoplasmic staining with high staining in bronchial ciliary rootlets and Sertoli cells, while accounting for cell-type variation (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AGPS 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 | Granular cytoplasm (HPA tissue IHC); peroxisomal association expected (UniProt) | |
| Staining pattern | Granular cytoplasm, most abundant in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03481-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03481-1) | |
| Caveat | Bronchial ciliary rootlets stain strongly, while alveolar cells may not (HPA tissue IHC) | |
| Regulation | Low tissue specificity; no regulator specified (HPA tissue IHC; UniProt) | |
| Isoform / epitope | No isoforms annotated; the mature chain spans residues 59–658 (UniProt) |
Section 1
Recommended AGPS IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A03481-1) is accompanied by published AGPS methods for human renal biopsies (PMC12460175) and prostate tissues (PMC10782585).
| Sample | Paraffin-embedded human adenocarcinoma of lung tissue; fixative not specified (datasheet A03481-1) |
| Fixation | Image fixative and duration unreported (datasheet A03481-1); 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 A03481-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03481-1) |
| Primary antibody | Rabbit anti-AGPS, 2-5 μg/ml (datasheet A03481-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03481-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03481-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AGPS-positive staining in ciliated cells (ciliary rootlets) of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous granular cytoplasmic expression, most abundant in glandular cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A03481-1); neither published method specifies retrieval (PMC12460175; PMC10782585).
Section 2
What Is the Expected AGPS Staining Pattern?
AGPS is associated with peroxisomes and the peroxisome membrane, with no predicted transmembrane segment (UniProt O00116 topology). In tissue IHC, expect granular cytoplasmic staining across many cell types, particularly glandular cells; strong staining is reported in selected ciliated and Sertoli cells (HPA: tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression (HPA: reliability).
| Discrete cytoplasmic granules in glandular cells, with little nuclear staining. | This fits the reported ubiquitous granular cytoplasmic tissue pattern and peroxisomal location (HPA: tissue IHC; HPA: subcellular; UniProt O00116). Score the stained cell type and intensity alongside the compartment; a granular pattern alone does not establish antibody specificity. |
| Strong, spatially restricted signal at ciliary rootlets in bronchus or fallopian tube. | High staining is reported specifically in ciliated cells at their ciliary rootlets (HPA: bronchus and fallopian tube IHC). Record that location separately from general cytoplasmic granules; the expected appearance depends on which cells are present in the section. |
| Predominantly nuclear, uniform membrane, or broad extracellular staining. | These locations depart from the reported granular cytoplasmic IHC pattern and peroxisomal localization (HPA: tissue IHC and subcellular; UniProt O00116). Treat them as possible artefact or nonspecific detection, then assess controls before assigning a biological interpretation. |
| Strong signal in adipocytes, cardiomyocytes, or lung alveolar cells. | HPA reports AGPS as not detected in these cells (HPA: adipose tissue, heart muscle, and lung IHC). Unexpected staining raises cross-reactivity or endogenous detection activity as possibilities; it does not prove either cause. Compare the signal with a matched negative detection control. |
| No staining in an adequately represented positive control tissue. | Check the relevant cells before calling the assay negative: HPA reports high signal in testis Sertoli cells and ciliated rootlets, and medium signal in several glandular tissues (HPA: tissue IHC). Missing signal warrants review of the IHC workflow and control performance. |
💡Expected AGPS appearanceCall a positive when the appropriate cells show granular cytoplasmic AGPS signal, potentially high in ciliary rootlets or Sertoli cells (HPA: tissue IHC); dominant nuclear or diffuse extracellular staining is discordant with the reported location (HPA: tissue IHC and subcellular).
| Subcellular location | AGPS is assigned to peroxisomes and the peroxisome membrane without a transmembrane segment (UniProt O00116 topology); HPA reports peroxisomes as the enhanced ICC-IF location (HPA: subcellular). Interpret cytoplasmic puncta in that context, while avoiding organelle identification from chromogenic IHC alone. |
| Choice of tissue and cells | HPA describes ubiquitous granular cytoplasmic expression, most abundant in glandular cells, yet reports cell-specific levels ranging from high to not detected (HPA: tissue IHC). Choose controls by the documented cell population, and score cells within the tissue rather than treating the whole section as uniformly positive. |
| Strength of IHC evidence | The tissue profile is Supported, with medium antibody–RNA consistency; HPA030209, HPA030210, and HPA030211 each have Supported IHC status (HPA: tissue reliability and antibody validation). This supports the reported pattern but leaves unexpected staining for control-based review. |
| Retrieval and epitope information | The supplied UniProt and HPA records give no antibody epitope or target-specific retrieval response. If signal is weak, compare retrieval conditions using the IHC-validated antibody's instructions and positive controls as a general IHC optimization step; do not infer an AGPS-specific fixation effect. |
| IF/ICC Q&A: what signal is expected? | Expect peroxisomal puncta: HPA lists peroxisomes as the enhanced ICC-IF location and reports enhanced ICC validation for HPA030209 and HPA030210 (HPA: subcellular and antibody validation). This informs localization only; the staining conditions belong in the separate IF/ICC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive section has no detectable chromogenic signal. | The relevant cells may be absent from the sampled area, or an IHC step may have failed; HPA reports high staining in Sertoli cells and ciliated rootlets (HPA: tissue IHC). | Confirm those cells are present, then check positive and negative controls, antibody dilution, retrieval, and detection reagents as general IHC practice. Do not infer AGPS-specific fixation sensitivity from the HPA pattern. |
| Granular signal is weak in liver, pancreas, or cerebral cortex. | Low staining is reported in hepatocytes, exocrine pancreatic cells, and cortical neurons (HPA: tissue IHC). Weak signal there can fit the reference pattern. | Compare the relevant cell type with a documented stronger control, such as Sertoli cells or ciliated rootlets (HPA: tissue IHC), before increasing detection strength or scoring the sample as negative. |
| Brown deposit is widespread, diffuse, or present where AGPS is reported as undetected. | Background, endogenous detection activity, or antibody cross-reactivity are plausible IHC causes; HPA reports not detected staining in several specified cell types (HPA: tissue IHC). | Inspect a matched negative detection control and review blocking, washes, and detection chemistry as general IHC practice. Reassess whether signal is confined to cells and compartments consistent with HPA. |
| Nuclei stain more strongly than the cytoplasm. | A nuclear-dominant pattern conflicts with granular cytoplasmic IHC and peroxisomal localization (HPA: tissue IHC and subcellular; UniProt O00116). | Check whether counterstain or nonspecific detection explains the appearance using controls. Report the nuclear pattern as discordant unless independent evidence supports its identity. |
| A gland-rich section appears uniformly negative. | HPA reports the strongest overall abundance in glandular cells but specifies medium staining for appendix, colon, duodenum, and endometrium glandular cells (HPA: tissue IHC). | Verify that intact glandular cells are represented and compare them with a documented high-signal control (HPA: tissue IHC). Review the general IHC workflow before interpreting the absence as biological. |
| Two antibodies give different IHC intensity or distribution. | HPA assigns Supported IHC status to HPA030209, HPA030210, and HPA030211; the tissue profile has only medium antibody–RNA consistency (HPA: antibody validation and tissue reliability). | Score each antibody's cellular and subcellular pattern against the same positive and negative tissue controls (HPA: tissue IHC). Document discordance instead of assuming that either intensity reflects AGPS abundance. |
Experiment evidence
IHC & IF images showing AGPS staining
A03481-1 · IHCIHC analysis of AGPS using anti-AGPS antibody (A03481-1). AGPS was detected in a paraffin-embedded section of human adenocarcinoma of lung t
A03481-1 · IHCIHC analysis of AGPS using anti-AGPS antibody (A03481-1). AGPS was detected in a paraffin-embedded section of human adenocarcinoma of lung t
A03481-1 · IFIF analysis of AGPS using anti-AGPS antibody (A03481-1). AGPS was detected in an immunocytochemical section of SiHa cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000018510-AGPS/tissue/cerebral+cortex#imid_8049699
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000018510-AGPS/tissue/colon#imid_8042057
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000018510-AGPS/tissue/liver#imid_8042058
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000018510-AGPS/tissue/kidney#imid_8042059
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA030209; green = AGPS, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA030209; green = AGPS, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AGPS IHC & IF
🧪Run bronchus first and look for staining in ciliated cells at the ciliary rootlets (HPA: High in bronchus ciliated cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the bronchus slide, assess neighboring nonciliated cells for background without assuming they are AGPS-negative.
Positive control tissue: Bronchus (Ciliated cells (ciliary rootlets), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AGPS in A-431, U-251MG, U2OS, with annotated localisation: Peroxisomes (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody), plus an AGPS knockout specimen if available. Quench endogenous peroxidase and check for residual signal in bronchus before interpreting DAB staining (selected-SKU caption: peroxidase detection with DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). The demonstrated IHC procedure uses heat retrieval in EDTA at pH 8.0, but a retrieval dependency has not been established; the supplied evidence does not show whether frozen sections or IF would be easier (selected-SKU caption: EDTA heat retrieval; HPA: ICC-IF images). In bronchus, distinguish the reported ciliary-rootlet pattern from nonspecific apical staining when scoring (HPA: High in ciliated cells at ciliary rootlets).
HPA tissue IHC evidence for AGPS
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 |
|---|---|---|---|---|
| Bronchus | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Testis | Sertoli cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AGPS IHC Tips
Troubleshoot AGPS staining in paraffin sections by checking retrieval, cellular pattern, and controls before comparing signal across specimens.
What retrieval should I start with for AGPS in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03481-1). In the documented tissue image, this preceded 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption A03481-1). If staining is weak, compare shorter and longer heat exposures on adjacent sections while holding antibody concentration, development time, and detection chemistry constant (standard IHC practice). Include a positive section and a no-primary control so increased background is not mistaken for recovered signal; the reported AGPS pattern is granular cytoplasmic and peroxisomal (HPA tissue IHC; HPA subcellular; standard IHC practice).
How should I assess whether fixation affects AGPS staining?
The paraffin-section caption does not report a fixative, so AGPS-specific fixation sensitivity is unknown (caption A03481-1). For a fixation comparison, process matched specimens under each condition and keep EDTA retrieval at pH 8.0, 2 μg/ml primary antibody, and overnight incubation at 4°C constant (caption A03481-1; standard IHC practice). Score granular cytoplasmic signal alongside preserved tissue architecture and no-primary background, rather than treating every increase in brown stain as improved detection (HPA tissue IHC; standard IHC practice). Record fixation duration and any delay before fixation for each specimen when comparing staining intensity (standard IHC practice).
Where should convincing AGPS staining appear?
Expect granular cytoplasmic staining compatible with peroxisomes, rather than a dominant nuclear pattern (HPA tissue IHC; HPA subcellular). UniProt annotates AGPS to peroxisomes and the peroxisome membrane but lists 0 transmembrane segments, so a crisp plasma-membrane rim is unsupported (UniProt O00116 localisation/topology). Assess granules within intact cells at consistent magnification and compare sections processed with the same EDTA retrieval at pH 8.0 (datasheet A03481-1; standard IHC practice). If DAB granules are difficult to resolve, compare an independent peroxisomal marker on a separate or multiplex section; brightfield proximity alone does not establish colocalisation (standard IHC practice).
Could processing or epitope position explain variable AGPS staining?
UniProt lists 0 AGPS isoforms and annotates a processed chain spanning residues 59–658 (UniProt O00116 isoforms/processing). The supplied antibody caption does not identify its epitope, so staining differences cannot be assigned to recognition of the precursor or processed chain (caption A03481-1; UniProt O00116 processing). Annotated modifications include phosphoserine 65 and phosphothreonine 74, but their effect on antibody binding is unreported (UniProt O00116 modified residues; caption A03481-1). If specimens stain differently, first compare EDTA retrieval at pH 8.0, section handling, and controls before proposing an epitope-specific explanation (datasheet A03481-1; standard IHC practice).
How can IF help check the AGPS pattern seen by IHC?
For a separate IF experiment, pair AGPS with a validated ciliated-cell marker in bronchus, where ciliated cells show high AGPS staining at ciliary rootlets (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and assess tissue autofluorescence in an unstained control; a far-red channel can help when shorter wavelengths are bright (standard IF practice). AGPS has 0 annotated transmembrane segments, but the catalog antibody’s epitope and its accessible side are unreported; compare permeabilisation conditions rather than assuming access (UniProt O00116 topology; caption A03481-1; standard IF practice). Judge puncta against a peroxisomal marker, since HPA reports peroxisomal localisation (HPA subcellular; standard IF practice).
How can I reduce diffuse brown staining without losing AGPS puncta?
Start by comparing the stained section with no-primary and reagent-only controls, then check whether diffuse colour follows tissue edges or damaged regions (standard IHC practice). The documented paraffin-section workflow used 10% goat serum, 2 μg/ml primary antibody, an HRP secondary, and DAB (caption A03481-1). If background persists, titrate primary antibody and DAB development while keeping EDTA retrieval at pH 8.0 and control processing matched (datasheet A03481-1; standard IHC practice). Include an endogenous-peroxidase blocking step and check its effect with controls; the reported AGPS pattern is granular cytoplasmic, so uniform haze should not be scored as signal (HPA tissue IHC; standard IHC practice).
How should I quantify AGPS staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, because HPA reports cell-dependent staining, including high signal in Sertoli cells and no detected signal in alveolar cells (HPA tissue IHC; standard IHC practice). Within each region, report the percentage of positive evaluable cells and an intensity-based H-score from 0–300, using the same thresholds across specimens (standard IHC practice). Alternatively, measure positive-cell density per mm² and normalise to viable tissue area, excluding necrosis and section artefacts (standard IHC practice). Record retrieval, antibody concentration, DAB development, and batch, and score granular cytoplasmic staining separately from background (caption A03481-1; HPA tissue IHC; standard IHC practice).
What distinguishes genuine AGPS staining from artefact?
A plausible positive has granular cytoplasmic staining in an appropriate cell population, consistent with HPA’s tissue pattern and peroxisomal localisation (HPA tissue IHC; HPA subcellular). High staining in bronchial ciliated cells or Sertoli cells can support the run, whereas HPA reports AGPS as not detected in alveolar cells (HPA tissue IHC). Treat dominant nuclear colour, edge staining, and staining confined to necrotic areas as suspect, and compare each with matched no-primary controls (HPA subcellular; standard IHC practice). Check endogenous peroxidase and DAB background before interpreting unexpected brown deposits, particularly when using the documented HRP and DAB workflow (caption A03481-1; standard IHC practice).
Boster reagents
Best AGPS / Alkyldihydroxyacetonephosphate synthase, peroxisomal IHC Antibodies
A03481-1 has IHC images from human paraffin sections and an IF image from SiHa cells (catalog image captions).
Real IHC data
Anti-AGPS Antibody ®
Cat # A03481-1
A03481-1 was used for IHC on paraffin sections of human lung adenocarcinoma, cervical cancer and esophageal squamous cell carcinoma (catalog IHC image captions). The same SKU was used for IF on SiHa cells (catalog IF image caption).
Which to pick: For tissue IHC, choose A03481-1 at 2–5 μg/ml (catalog IHC dilution); its image captions show paraffin sections with EDTA retrieval, while the fixative is unreported (catalog IHC image captions). For IF/ICC, choose A03481-1 at 5 μg/ml (catalog applications and IF dilution); its IF image shows SiHa cells (catalog IF image caption). For cross-species work, this catalog offers no validated option beyond human reactivity (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00116 (ADAS_HUMAN, Alkyldihydroxyacetonephosphate synthase, peroxisomal).
- Human Protein Atlas. AGPS tissue IHC expression (reliability: Supported).
- Human Protein Atlas. AGPS subcellular location (ICC-IF): Localized to the peroxisomes..
- Human Protein Atlas. AGPS antibody validation summary (3 antibodies).
- Multiple oestradiol functions inhibit ferroptosis and acute kidney injury. Nature 2025 — PMC12460175.
- Alkylglycerone phosphate synthase (AGPS) deficient mice: models for rhizomelic chondrodysplasia punctate type 3 (RCDP3) malformation syndrome. Molecular genetics and metabolism reports 2014 — PMC4151185.
- TrkA promotes MDM2-mediated AGPS ubiquitination and degradation to trigger prostate cancer progression. Journal of experimental & clinical cancer research : CR 2024 — PMC10782585.
- PubMed PMID:9187299 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AGPS (Alkyldihydroxyacetonephosphate synthase, peroxisomal). Expected localisation Granular cytoplasm (HPA tissue IHC); peroxisomal association expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03481-1). Validated IHC antibody: A03481-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (ciliary rootlets)); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human adenocarcinoma of lung tissue; fixative not specified (datasheet A03481-1); 2. Fixation: Image fixative and duration unreported (datasheet A03481-1); 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 A03481-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03481-1); 8. Primary antibody: Rabbit anti-AGPS, 2-5 μg/ml (datasheet A03481-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03481-1). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03481-1); Caveat: Bronchial ciliary rootlets stain strongly, while alveolar cells may not (HPA tissue IHC); Isoform / epitope: No isoforms annotated; the mature chain spans residues 59–658 (UniProt); Regulation: Low tissue specificity; no regulator specified (HPA tissue IHC; UniProt). Evidence: UniProt O00116, HPA tissue IHC · Supported, HPA ICC-IF · peroxisomes., Datasheet A03481-1, PMC12460175, PMC10782585. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AGPS/immunohistochemistry-ihc-guide.