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ISYNA1 / Inositol-3-phosphate synthase 1 · IHC design guide
Design Immunohistochemistry for ISYNA1
Plan ISYNA1 IHC in paraffin sections using its cytoplasmic tissue staining profile (HPA tissue IHC). Use the catalog antibody’s IHC conditions and compare staining in defined cell populations (datasheet A08693-1; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ISYNA1 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 (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in late spermatids and cytotrophoblasts (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08693-1) | |
| Positive control | Fallopian tube+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 A08693-1) | |
| Caveat | Staining varies among cell types within tissues (HPA tissue IHC) | |
| Regulation | Tissue-dependent expression (UniProt) | |
| Isoform / epitope | 3 isoforms; antibody epitope coverage is unspecified (UniProt; datasheet A08693-1) |
Section 1
Recommended ISYNA1 IHC & IF Protocols
The catalog antibody protocol and two published ISYNA1 IHC reports cover paraffin sections from breast metastasis and pancreatic cancer studies (PMC13557034; PMC7521282).
| Sample | Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A08693-1) |
| Fixation | Image fixative and duration unreported (datasheet A08693-1); 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 A08693-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08693-1) |
| Primary antibody | Rabbit anti-ISYNA1, 2-5 μg/ml (datasheet A08693-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08693-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08693-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ISYNA1-positive staining in non-ciliated cells of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several different tissue types, most abundant in testis and placental trophoblast. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A08693-1); neither included article specifies a retrieval condition.
Section 2
What Is the Expected ISYNA1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic ISYNA1 staining (UniProt Q9NPH2: cytoplasm; HPA: cytoplasmic tissue expression). The clearest reported cells include placental cytotrophoblasts, late spermatids, fallopian tube non-ciliated cells and cells of Bowman's capsule (HPA: High). ISYNA1 has no transmembrane segment (UniProt Q9NPH2: topology). HPA rates the tissue pattern Enhanced for consistency with RNA expression, pending external verification (HPA: reliability).
| Distinct cytoplasmic staining in placental cytotrophoblasts or testicular late spermatids. | This fits the reported High cell-specific tissue pattern (HPA: placenta and testis). Compare cells within the section; a positive tissue need not stain uniformly (HPA: cell-specific tissue IHC). |
| Predominantly nuclear staining in a paraffin section, with little cytoplasmic staining. | Check IHC controls and scoring before calling this the expected tissue pattern (HPA: cytoplasmic tissue profile). Nuclear signal alone is not proof of artefact: ICC-IF reports approved nucleoplasmic localization (HPA: subcellular). |
| Strong staining in a cell population reported as not detected, such as adipocytes. | Treat this as a specificity warning, especially if staining is widespread (HPA: adipocytes not detected). Cross-reactivity or endogenous detection activity are possible explanations (general IHC practice); review controls before assigning ISYNA1 expression. |
| Diffuse chromogen across cells and surrounding tissue, without clear cell boundaries. | This cannot establish the reported cytoplasmic, cell-specific pattern (HPA: tissue IHC). Background from detection reagents or nonspecific binding is possible (general IHC practice); compare a no-primary control. |
| No staining in placental cytotrophoblasts or testicular late spermatids. | A negative result conflicts with their reported High staining (HPA: placenta and testis). Check the run and section quality before interpreting other tissues as negative (general IHC practice). |
💡Expected ISYNA1 appearanceA convincing positive is distinct, predominantly cytoplasmic chromogen in reported High cells, such as placental cytotrophoblasts (HPA: tissue IHC); diffuse staining or signal in no-primary controls is suspect (general IHC practice).
| Cell identity within tissue | Interpret the stained cell population, not just the organ: Bowman's capsule is High, while pancreatic exocrine glandular cells are Low (HPA: tissue IHC). |
| Subcellular localization | Cytoplasmic tissue staining agrees with UniProt's cytoplasm assignment and absence of a transmembrane segment (HPA: tissue IHC; UniProt Q9NPH2: location and topology). |
| IF/ICC Q: Can nuclear signal be expected? | A: Yes in ICC-IF: HPA approves nucleoplasm as the main location and cytosol as an additional location (HPA: subcellular). Evaluate paraffin IHC against its separate cytoplasmic tissue profile (HPA: tissue IHC). |
| Strength of tissue evidence | HPA labels tissue IHC Enhanced and lists 3 antibodies with Enhanced IHC status (HPA: reliability and antibodies). The tissue profile still awaits external verification (HPA: reliability description). |
| Antigen retrieval and fixation | No ISYNA1-specific fixation sensitivity or retrieval condition is supplied (UniProt Q9NPH2; HPA: supplied records). Assess retrieval as a general IHC workflow variable, without predicting a target-specific effect (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank. | A run failure, unsuitable section or suboptimal staining conditions may suppress detection (general IHC practice); these cells are reported High (HPA: placenta and testis). | Check a known-positive section in the same run, then review the catalog antibody's IHC-P instructions, detection reagents and section quality (general IHC practice). |
| All tissue compartments show similar brown signal. | Nonspecific binding or endogenous detection activity may obscure cell-specific staining (general IHC practice; HPA: tissue IHC profile). | Compare a no-primary control; review blocking, washes and detection chemistry as general IHC steps (general IHC practice). |
| Signal is mainly nuclear in tissue IHC. | This differs from HPA's cytoplasmic tissue profile, although HPA ICC-IF approves nucleoplasmic localization (HPA: tissue IHC and subcellular). | Record assay and cell context; check controls and cytoplasmic signal before assigning the nuclear staining to ISYNA1 (general IHC practice). |
| A reported negative cell population stains strongly. | Cross-reactivity or endogenous chromogen activity is possible (general IHC practice); HPA reports adipocytes as not detected (HPA: adipose tissue). | Check a no-primary control and compare the population with a reported positive on the same run (general IHC practice; HPA: tissue IHC). |
| Only faint staining appears in pancreatic exocrine cells. | Low staining is reported there (HPA: pancreas), so faint signal alone does not show that the run failed. | Judge sensitivity using reported High cells, such as placental cytotrophoblasts, and retain cell-level scoring (HPA: placenta and pancreas). |
| ICC-IF and paraffin IHC appear to disagree. | HPA reports mainly nucleoplasmic ICC-IF localization but cytoplasmic tissue IHC staining (HPA: subcellular and tissue IHC). | Interpret each assay against its own HPA profile; document the cell type and compartment instead of transferring an ICC-IF expectation directly to tissue IHC (HPA: both profiles). |
Experiment evidence
IHC & IF images showing ISYNA1 staining
A08693-1 · IHCIHC analysis of ISYNA1 using anti-ISYNA1 antibody (A08693-1). ISYNA1 was detected in a paraffin-embedded section of human glioblastoma tissu
A08693-1 · IHCIHC analysis of ISYNA1 using anti-ISYNA1 antibody (A08693-1). ISYNA1 was detected in a paraffin-embedded section of human liver cancer tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105655-ISYNA1/tissue/cerebral+cortex#imid_19794801
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105655-ISYNA1/tissue/colon#imid_19788671
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105655-ISYNA1/tissue/liver#imid_19788672
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000105655-ISYNA1/tissue/kidney#imid_19788689
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007931; green = ISYNA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007931; green = ISYNA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ISYNA1 IHC & IF
🧪Run placenta first: cytotrophoblasts should stain strongly (HPA: High in placental cytotrophoblasts). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); on the placenta slide, score morphologically distinct cells without specific staining as internal negatives only if such cells are present.
Positive control tissue: Fallopian tube (Non-ciliated cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ISYNA1 in A-431, HEK293, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-matched rabbit control IgG matched to the catalog antibody’s clonality, plus ISYNA1 knockout material or immunogen-peptide competition as a biological specificity control (hero caption: rabbit anti-ISYNA1 antibody; standard IHC practice). On placenta, check blood-associated endogenous peroxidase background after peroxidase blocking, using the no-primary slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected A08693-1 paraffin-section caption also does not state a fixative (hero caption: fixative not stated). The documented IHC procedure uses heat-mediated EDTA retrieval at pH 8.0, but the evidence does not establish that retrieval is required for every specimen (hero caption: EDTA retrieval, pH 8.0). Neither frozen sections nor IF are shown to be easier by the supplied evidence; placental blood-associated peroxidase can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for ISYNA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Fallopian tube | Non-ciliated cells | High | Protein (IHC) | HPA → |
| Kidney | Bowman's capsule | High | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ISYNA1 IHC Tips
Troubleshoot chromogenic ISYNA1 IHC in paraffin sections using the catalog antibody’s tissue image, protein annotations, and tissue staining patterns.
Which retrieval conditions should I start with for ISYNA1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A08693-1). The reported paraffin-section image used that retrieval before 2 μg/ml primary antibody overnight at 4°C (datasheet A08693-1). If staining is weak, vary heating duration while keeping the buffer and antibody concentration fixed, and compare a positive control on the same run (HPA: high staining in placental cytotrophoblasts). Assess tissue integrity alongside signal: excessive heating can damage morphology and make cytoplasmic staining difficult to score. Record retrieval time and instrument settings so that a stronger signal can be distinguished from variation between runs.
Could fixation explain weak or patchy ISYNA1 staining?
Target-specific fixation sensitivity is unknown: the catalog image describes a paraffin section but does not state its fixative (datasheet A08693-1). Record the actual fixative, fixation duration, and time before processing for each specimen; compare sections processed together when investigating patchiness. In formalin-fixed material, incomplete or prolonged fixation can change antigen accessibility as a general IHC concern, so assess morphology and retrieval performance before changing antibody concentration. Use a concurrently processed positive control with an expected staining pattern, such as placental cytotrophoblasts (HPA: High), and avoid attributing a failed stain specifically to ISYNA1 fixation sensitivity without a controlled comparison.
Should I score nuclear staining as ISYNA1 positive in tissue IHC?
Score the compartment explicitly rather than combining nuclear and cytoplasmic signal: tissue IHC shows cytoplasmic expression across several tissues (HPA: tissue IHC profile). UniProt places ISYNA1 in the cytoplasm, whereas cell imaging reports mainly nucleoplasmic localisation with additional cytosol (UniProt Q9NPH2; HPA: subcellular localisation). Nuclear staining therefore deserves separate review, particularly when cytoplasmic signal is absent in an expected positive population. Compare adjacent morphology, a no-primary control, and a known positive tissue under identical detection conditions; report nuclear and cytoplasmic scores separately until the nuclear pattern is supported in the tissue assay.
How could isoforms or epitope accessibility affect my tissue staining?
ISYNA1 has 3 annotated isoforms, but the supplied evidence does not map the catalog antibody’s epitope to any one of them (UniProt Q9NPH2). Its annotated chain spans residues 1–558, with phosphoserines at 279 and 357; epitope overlap with either site is unknown (UniProt Q9NPH2). Treat unexpected staining differences as an epitope-accessibility question only after checking section processing, retrieval, and positive-control performance. For isoform-specific interpretation, obtain the antibody’s immunogen or epitope sequence and compare it with isoform sequences; total chromogenic staining alone cannot establish which isoform contributed signal.
How should I investigate ISYNA1 by multiplex IF after tissue IHC?
For multiplex IF, pair ISYNA1 with a validated marker of the expected cell population, such as placental cytotrophoblasts, and assess overlap at single-cell resolution (HPA: High in placental cytotrophoblasts). Choose a fluorophore channel with low tissue autofluorescence, often a longer-wavelength channel, and include unstained and single-stain controls to check bleed-through. ISYNA1 has no annotated transmembrane segment, so permeabilise cells to access intracellular epitopes while comparing conditions that preserve nuclear and cytosolic morphology (UniProt Q9NPH2). Cell imaging reports nucleoplasm and cytosol, but that observation does not establish IF performance for catalog antibody A08693-1 (HPA: subcellular localisation; datasheet A08693-1).
What should I change when ISYNA1 DAB staining is diffuse?
Check a no-primary section first to separate secondary-detection or endogenous-peroxidase signal from primary-antibody staining. The catalog image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB detection (datasheet A08693-1). In the general chromogenic workflow, block endogenous peroxidase before the HRP step, then review wash quality and DAB development time. Compare background in expected low or undetected populations with a positive region on the same run; adipocytes were not detected in the cited tissue profile, but that result alone does not validate every adipose specimen as a negative control (HPA: adipocytes Not detected).
How can I quantify ISYNA1 IHC across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; tissue IHC reports cytoplasmic expression, while cell imaging also reports nucleoplasmic signal (HPA: tissue IHC profile; HPA: subcellular localisation). For chromogenic staining, record the percentage of positive target cells and an intensity score, or calculate an H-score from intensity categories 0–3 and their cell percentages. Normalise positive-cell counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counting is impractical. Apply identical thresholds, counterstain assessment, and region-selection rules across slides, and exclude folds, edges, and necrotic areas before comparing specimens.
How do I distinguish true ISYNA1 staining from artefact?
Look for reproducible signal in intact cells with an expected compartment and cell distribution: ISYNA1 is annotated as cytoplasmic, and tissue IHC is predominantly cytoplasmic (UniProt Q9NPH2; HPA: tissue IHC profile). Placental cytotrophoblasts and late spermatids offer reported high-staining reference populations, whereas the catalog image demonstrates staining in a paraffin section of human glioblastoma (HPA: High; datasheet A08693-1). Treat staining confined to cut edges, folds, necrosis, or morphologically implausible cells as suspect and compare it with a no-primary control. If DAB appears in the control, assess endogenous peroxidase and detection reagents before interpreting the signal as ISYNA1.
Boster reagents
Best ISYNA1 / Inositol-3-phosphate synthase 1 IHC Antibodies
A08693-1 has real IHC data from human paraffin sections (IHC image captions); its listed IHC reactivity includes human, mouse and rat (catalog: reactivity). No IF data are supplied (catalog: IF images).
Real IHC data
Anti-ISYNA1 Antibody ®
Cat # A08693-1
A08693-1 is listed for IHC in human, mouse and rat (catalog: applications and reactivity). Its IHC captions show human paraffin sections of glioblastoma, liver cancer, lung cancer and pancreatic ductal adenocarcinoma (IHC image captions).
Which to pick: Choose A08693-1 for paraffin-section IHC: its image captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (IHC image captions). It is also listed as reactive with mouse and rat for IHC, although the supplied images show human tissue (catalog: reactivity; IHC image captions). No IF/ICC application or image is supplied, so there is no supported IF/ICC pick here; the fixative used for the pictured sections is unreported (catalog: applications and IF images; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NPH2 (INO1_HUMAN, Inositol-3-phosphate synthase 1).
- Human Protein Atlas. ISYNA1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ISYNA1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. ISYNA1 antibody validation summary (3 antibodies).
- Proteomics analysis of melanoma metastases: association between S100A13 expression and chemotherapy resistance. British journal of cancer 2014 — PMC4021518.
- Integrative Transcriptional and Chromatin Analyses Reveal Enhancer-Mediated Regulatory Programs Driving Breast Cancer Metastasis. Molecular cancer research : MCR 2026 — PMC13557034.
- Colony-stimulating factor 1 positive (CSF1(+) ) secretory epithelial cells induce excessive trophoblast invasion in tubal pregnancy rupture. Cell proliferation 2023 — PMC10334268.
- Musashi2 promotes the progression of pancreatic cancer through a novel ISYNA1-p21/ZEB-1 pathway. Journal of cellular and molecular medicine 2020 — PMC7521282.
- PubMed PMID:12941308 — UniProt-cited evidence.
- PubMed PMID:17121280 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ISYNA1 (Inositol-3-phosphate synthase 1). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08693-1). Validated IHC antibody: A08693-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Non-ciliated cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A08693-1); 2. Fixation: Image fixative and duration unreported (datasheet A08693-1); 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 A08693-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08693-1); 8. Primary antibody: Rabbit anti-ISYNA1, 2-5 μg/ml (datasheet A08693-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08693-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08693-1); Caveat: Staining varies among cell types within tissues (HPA tissue IHC); Isoform / epitope: 3 isoforms; antibody epitope coverage is unspecified (UniProt; datasheet A08693-1); Regulation: Tissue-dependent expression (UniProt). Evidence: UniProt Q9NPH2, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08693-1, PMC13557034, PMC7521282. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ISYNA1/immunohistochemistry-ihc-guide.