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- Table of Contents
Plan PTGES2 IHC around granular cytoplasmic tissue staining (HPA tissue IHC) and an antibody starting concentration of 2–5 μg/ml (datasheet A06706-2). Compare high staining in kidney tubules with low staining in hippocampal neurons (HPA tissue IHC), while accounting for cleavage from a Golgi-bound to a cytoplasmic form (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Granular cytoplasm (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06706-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Soft tissue |
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Cleavage can shift signal from Golgi to cytoplasm (UniProt) | |
| Regulation | Tissue-dependent expression (UniProt) | |
| Isoform / epitope | No isoforms; cleavage removes aa 1–87, retaining aa 88–377 (UniProt) |
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A06706-2). Two published PTGES2 IHC protocols provide additional conditions for renal carcinoma and endometrial specimens (PMC12484182; PMC5080328).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06706-2) |
| Fixation | Image fixative and duration unreported (datasheet A06706-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A06706-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06706-2) |
| Primary antibody | Rabbit anti-PTGES2, 2-5 μg/ml (datasheet A06706-2) |
| Primary incubation | Overnight at 4 °C (datasheet A06706-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06706-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PTGES2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
PTGES2 IHC should show mainly granular cytoplasmic staining in appropriate cells (HPA tissue IHC: Approved; medium consistency with RNA). A perinuclear emphasis is plausible because the predominant cleaved form is cytoplasmic and enriched there; the precursor is Golgi membrane bound, with one transmembrane segment at residues 58–74 (UniProt Q9H7Z7: processing and topology). Judge the pattern within each tissue’s documented cell type, rather than treating every cell as equally positive (HPA tissue IHC).
| Granular cytoplasmic signal, sometimes concentrated around the nucleus, in kidney tubule cells or heart cardiomyocytes. | This fits the reported tissue pattern and High staining in those cell types (HPA tissue IHC). Perinuclear enrichment also fits the predominant soluble, cleaved form (UniProt Q9H7Z7: processing). Compare cell identity, compartment, and intensity together; a brown deposit alone is insufficient to establish a PTGES2-positive cell (general IHC interpretation). |
| Predominantly nuclear, sharply membranous, or extracellular deposit, with little granular cytoplasmic signal. | The dominant compartment conflicts with the reported granular cytoplasmic IHC profile (HPA tissue IHC) and the cytoplasmic, perinuclear cleaved form (UniProt Q9H7Z7: subcellular location). Review morphology and controls before calling it specific; the Golgi-bound precursor permits intracellular membrane-associated signal, but does not by itself explain strong surface or extracellular staining (UniProt Q9H7Z7: processing and topology). |
| Strong staining in peripheral nerve within soft tissue, or in cells other than those expected in a positive tissue. | Peripheral nerve was Not detected in the HPA soft-tissue example (HPA tissue IHC). Unexpected staining can reflect cross-reactivity or endogenous chromogen activity (general IHC practice); it is a reason to check controls and cell identity, not proof of either cause. Tissue context matters because PTGES2 expression is broad and staining differs among cell types (UniProt Q9H7Z7: tissue specificity; HPA tissue IHC). |
| Uniform haze, precipitate, or colour across multiple compartments and cell types. | This obscures the granular cytoplasmic pattern reported for PTGES2 (HPA tissue IHC). Assess reagent-only and negative-control sections for detection background, incomplete blocking, or chromogen deposit (general IHC practice). Do not score diffuse colour as target expression solely because the specimen contains an HPA-listed positive cell type (HPA tissue IHC). |
| No staining in kidney tubule cells or heart cardiomyocytes on an otherwise interpretable section. | Both are reported as High IHC cell populations, so an absent signal raises concern about assay performance (HPA tissue IHC). Check the positive control, antibody use, retrieval, and detection steps (general IHC practice). HPA’s Approved rating has medium consistency with RNA, so one negative section should not be treated as definitive evidence that the target is absent (HPA tissue IHC). |
| Processing and compartment | PTGES2 is synthesized as a Golgi membrane-bound 1–377 protein and cleaved to a predominant soluble 88–377 form enriched in perinuclear cytoplasm (UniProt Q9H7Z7: chains and subcellular location). Interpret mixed intracellular staining in that context; these records do not give the catalog antibody’s epitope or prove which form its IHC signal detects. |
| Cell-type selection | HPA reports High staining in adrenal and endometrial glandular cells, kidney tubule cells, cardiomyocytes, pancreatic exocrine cells, parathyroid and rectal glandular cells, and Leydig cells (HPA tissue IHC). It reports Low staining in several other cell types, so match intensity to the cell population when choosing a comparison section (HPA tissue IHC). |
| Strength of validation | The tissue IHC profile is Approved, with medium consistency between antibody staining and RNA expression; HPA020733 is listed as IHC Approved (HPA tissue IHC; HPA antibodies). These labels support pattern comparison but do not establish that every brown cell is specific. No IHC Enhanced designation is supplied for this antibody (HPA antibodies). |
| Antigen retrieval and epitope | No retrieval condition or antibody epitope is supplied here. The transmembrane span is 58–74, while the cleaved chain begins at 88 (UniProt Q9H7Z7: topology and chains). Whether processing changes recognition depends on the unknown epitope; choose and assess retrieval with suitable controls as general IHC optimization, without assuming a PTGES2-specific fixation effect. |
| Chromogenic detection | Endogenous enzyme activity and nonspecific reagent binding can add colour in chromogenic IHC (general IHC practice). Use detection and negative controls to separate those signals from the reported granular cytoplasmic profile (HPA tissue IHC). The supplied HPA and UniProt records provide no target-specific claim about endogenous activity or fixation sensitivity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control tubules or cardiomyocytes show no signal. | A failed staining step is possible; these populations are High in HPA IHC (HPA tissue IHC). | Check control section quality, primary-antibody application, retrieval, and detection reagents (general IHC practice). Record the actual conditions tested; no PTGES2-specific retrieval setting is supplied. |
| Only nuclei stain strongly. | Nuclear dominance conflicts with the reported granular cytoplasmic pattern (HPA tissue IHC). | Check counterstain and negative-control sections, then reassess whether true cytoplasmic granules are present (general IHC practice). Do not score nuclear colour alone as PTGES2-positive. |
| Brown colour appears in peripheral nerve or many unexpected cells. | Peripheral nerve was Not detected in the HPA soft-tissue example; cross-reactivity or detection background is possible (HPA tissue IHC; general IHC practice). | Compare matched negative and detection controls, inspect morphology, and require the expected intracellular pattern before assigning positivity (general IHC practice). |
| Granular signal is buried under diffuse background. | Background can conceal the reported cytoplasmic granules (HPA tissue IHC; general IHC practice). | Review blocking, washes, chromogen development, and detection-only controls (general IHC practice). Adjust the step implicated by the control, then compare the same documented cell type. |
| A faint sample is being called negative. | Intensity varies by cell type; HPA reports Low staining in neuronal, squamous epithelial, salivary glandular, and smooth muscle cells (HPA tissue IHC). | Compare with a documented High cell population and the negative control on the same run (HPA tissue IHC; general IHC practice). Report faint, localized signal separately from complete absence. |
| Can IF/ICC confirm the IHC compartment? | HPA supplies no main ICC-IF location, cell-line images, or ICC validation entry for HPA020733 (HPA subcellular; HPA antibodies). | Treat IF/ICC as a separate assay requiring its own controls and validation (general IF practice). Use the IHC tissue pattern for IHC interpretation; no IF/ICC protocol is established by these records. |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Troubleshoot PTGES2 staining in paraffin sections by checking retrieval, controls, compartment, and cell type before interpreting chromogenic signal.
A06706-2 has real IHC data from paraffin-embedded human breast and ovarian cancer sections and IF data from Hela cells (catalog image captions). Listed reactivity includes human, mouse and rat (catalog reactivity).
A06706-2 shows IHC staining in paraffin-embedded human breast and ovarian cancer sections (catalog IHC captions). The same SKU shows IF staining in Hela cells and lists both IF and ICC applications (catalog IF caption; catalog applications).
Which to pick: For tissue IHC, choose A06706-2: its own images document paraffin-embedded human sections stained at 2 μg/ml; the fixative is unreported (catalog IHC captions). For IF/ICC, the same rabbit-host SKU lists both applications and shows IF staining in Hela cells at 5 μg/ml; clonality is unspecified (catalog host/clone; catalog applications; catalog IF caption). For mouse or rat samples, A06706-2 lists reactivity with both species, while its supplied IHC and IF images document human samples (catalog reactivity; catalog image captions).