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TPI1 / Triosephosphate isomerase · IHC design guide
Design Immunohistochemistry for TPI1
Plan TPI1 staining in paraffin sections using the IHC-validated antibody at 1:50–1:100 (datasheet). Compare the expected cytoplasmic location (UniProt) with the general nuclear and cytoplasmic tissue staining, which has uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TPI1 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 expected (UniProt); nuclear and cytoplasmic observed (HPA tissue IHC) | |
| Staining pattern | General nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Use formalin-fixed paraffin sections (selected-SKU IHC image A02559-1); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02559-1) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | RNA tissue-enhanced in muscle, tongue (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; C-terminal epitope coverage is unknown (UniProt; datasheet) |
Section 1
Recommended TPI1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published TPI1 tissue IHC method (PMC11531968).
| Sample | FFPE human prostate carcinoma tissue (datasheet A02559-1) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A02559-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-TPI1, 1:50-1:100 (datasheet A02559-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TPI1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule; UniProt P60174: cytoplasm).
Section 2
What Is the Expected TPI1 Staining Pattern?
TPI1 is a cytoplasmic enzyme with no transmembrane segment (UniProt P60174: subcellular location and topology). In paraffin IHC, assess intracellular staining in HPA-reported cells, including adipocytes, respiratory epithelium and glia (HPA: Medium). HPA also reports a general nuclear and cytoplasmic tissue pattern, while ICC-IF places TPI1 mainly in the nucleoplasm (HPA: tissue IHC; HPA: ICC-IF). Interpret compartment differences cautiously: HPA rates its tissue IHC evidence Uncertain (HPA: reliability).
| Intracellular staining in adipocytes, bronchial respiratory epithelium or caudate glia. | These are plausible positive cells: each has Medium tissue staining (HPA: tissue IHC). Compare a candidate positive section with a matched negative control before assigning TPI1 specificity (general IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic staining. | This differs from UniProt's cytoplasmic annotation (UniProt P60174: subcellular location), but nuclear staining is reported in tissue IHC and nucleoplasmic ICC-IF (HPA: tissue IHC; HPA: ICC-IF). Verify the pattern; nuclear signal alone is not proof of artefact. |
| Strong membrane-rim staining or staining confined to cell surfaces. | A membrane-dominant pattern conflicts with the cytoplasmic location and absence of a transmembrane segment (UniProt P60174: location and topology). Suspect nonspecific staining and check the matched negative control (general IHC practice). |
| Strong staining confined to a cell population outside the reported pattern. | Compare with HPA's cell-level observations, which vary by tissue (HPA: tissue IHC). Unexpected staining warrants investigation for antibody cross-reactivity or endogenous chromogen activity; it cannot be identified as TPI1 from appearance alone (general IHC practice). |
| Diffuse colour across cells and surrounding tissue, or no signal in a candidate positive section. | Neither result establishes TPI1 distribution. HPA reports Medium staining in selected cells but rates tissue IHC Uncertain (HPA: tissue IHC). Use controls to distinguish background or assay failure from a genuinely low signal (general IHC practice). |
💡Expected TPI1 appearanceCall a section provisionally positive when intracellular cytoplasmic and/or nuclear staining is discernible in HPA-reported cells, with approximately Medium staining in the listed positive populations (HPA: tissue IHC); a strong membrane rim or uniform diffuse deposit needs control-based review (UniProt P60174: topology; general IHC practice).
| How should the compartment calls be reconciled? | UniProt assigns TPI1 to the cytoplasm, whereas HPA describes nuclear and cytoplasmic tissue staining (UniProt P60174: location; HPA: tissue IHC). Score compartments separately; HPA's tissue reliability is Uncertain (HPA: reliability). |
| How should tissue and cell choice affect interpretation? | HPA records Medium staining in selected adipocytes, glandular cells, respiratory epithelial cells and glia, but Low staining in several other sampled populations (HPA: tissue IHC). Identify the scored cell type before comparing sections. |
| How strong is the IHC validation? | Both listed HPA antibodies, HPA050924 and HPA053568, have Uncertain IHC status (HPA: antibody validation). HPA attributes tissue uncertainty to low agreement between staining and RNA data, pending external verification (HPA: reliability). |
| Do isoforms or processing specify an expected staining pattern? | UniProt lists three isoforms and a chain spanning residues 2–249, with no signal peptide or propeptide (UniProt P60174: isoforms and processing). The supplied evidence gives no antibody epitope, so it cannot establish isoform-specific IHC staining. |
| Q: Can ICC-IF determine the expected paraffin IHC compartment? | A: HPA reports mainly nucleoplasmic ICC-IF staining and Approved ICC status for both listed antibodies (HPA: ICC-IF; HPA: antibody validation). Use that as context, while interpreting paraffin IHC against its own Uncertain tissue evidence (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No discernible signal in an HPA-listed Medium cell population. | The assay may have failed, or the sampled cells may differ from HPA's observation; tissue IHC reliability is Uncertain (HPA: tissue IHC). | Confirm cell identity, check the run's positive and negative controls, then review retrieval, primary-antibody dilution and detection conditions (general IHC practice). |
| Broad, pale chromogen haze obscures cell boundaries. | Nonspecific antibody binding or excessive detection signal can obscure compartment scoring (general IHC practice). | Review the matched negative control; optimise blocking, antibody dilution and detection time one variable at a time (general IHC practice). |
| Brown deposits persist in the negative control. | Endogenous peroxidase activity or nonspecific detection chemistry may contribute to chromogenic signal (general IHC practice). | Check the omitted-primary control and the detection system; apply an appropriate endogenous-peroxidase block if indicated (general IHC practice). |
| Signal is mainly at the cell membrane. | That distribution conflicts with TPI1's cytoplasmic location and lack of a transmembrane segment (UniProt P60174: location and topology). | Compare with controls and with intracellular staining in HPA-listed cells before calling the membrane signal TPI1 (HPA: tissue IHC; general IHC practice). |
| Nuclear staining appears stronger than cytoplasmic staining. | HPA reports nuclear tissue staining and mainly nucleoplasmic ICC-IF, despite UniProt's cytoplasmic annotation (HPA: tissue IHC; HPA: ICC-IF; UniProt P60174: location). | Record both compartments and check control staining. Treat a nuclear-only IHC result as provisional given HPA's Uncertain IHC reliability (HPA: reliability; general IHC practice). |
| A different cell type stains more strongly than the expected population. | HPA staining varies across sampled cell types, and its tissue IHC assessment is Uncertain (HPA: tissue IHC). Cross-reactivity or endogenous signal is also possible (general IHC practice). | Confirm the cell identities and inspect negative controls; report the observed distribution without assigning target specificity until supported (general IHC practice). |
Experiment evidence
IHC & IF images showing TPI1 staining
A02559-1 · IHCFormalin-fixed and paraffin-embedded human prostate carcinoma reacted with TPI1 Antibody (C-term), which was peroxidase-conjugated to the se
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000111669-TPI1/tissue/cerebral+cortex#imid_18369595
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000111669-TPI1/tissue/colon#imid_18369521
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000111669-TPI1/tissue/liver#imid_18369530
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000111669-TPI1/tissue/kidney#imid_18369523
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA050924; green = TPI1, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA050924; green = TPI1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TPI1 IHC & IF
🧪Run adipose tissue first and confirm staining in adipocytes (HPA: Medium in adipocytes). HPA detects TPI1 in all 45 scored tissues, so there is no supported negative tissue; use the no-primary and isotype controls as negatives, and treat unstained cells on the positive slide as a finding to investigate rather than established internal negative cells (HPA: no negative rows).
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: None in HPA: TPI1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TPI1 in HEK293, MCF-7, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Run a secondary-only slide and a host-species-matched isotype control for a monoclonal primary, or matched nonimmune IgG for a polyclonal primary; use TPI1 knockout material or a validated peptide-block control to test target specificity (standard IHC/IF practice). Quench endogenous peroxidase before chromogenic detection, and check adipose tissue for autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: The selected A02559-1 caption documents formalin-fixed, paraffin-embedded tissue with peroxidase/DAB detection, but gives no fixation duration or target-specific fixation window (selected A02559-1 caption). Retrieval dependence is unreported, so optimize retrieval empirically; the supplied evidence does not establish whether frozen sections or IF are easier for TPI1 (selected A02559-1 caption; HPA: ICC-IF images available). In adipose sections, lipid extraction during paraffin processing can affect morphology, so assess tissue preservation when scoring adipocytes (standard IHC practice).
HPA tissue IHC evidence for TPI1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: TPI1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TPI1 IHC Tips
Troubleshoot TPI1 staining in paraffin sections using the retrieval setting, the catalog antibody’s tissue image, and compartment-specific controls.
How should I retrieve TPI1 in paraffin sections with weak staining?
Start with citrate buffer at pH 6.0 and heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). TPI1 has no transmembrane segment and is annotated in the cytoplasm, so assess cytoplasmic staining alongside morphology after retrieval (UniProt P60174; standard IHC practice). If staining remains weak, compare a shorter or longer retrieval time on serial sections while keeping the antibody and detection conditions fixed (standard IHC practice). Record tissue damage and background as well as signal, because an apparently stronger stain is unhelpful when cellular boundaries become difficult to score (standard IHC practice).
Could fixation explain inconsistent TPI1 staining across paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied evidence, so treat block-to-block differences as a question to test experimentally (supplied evidence). The catalog antibody’s image shows formalin-fixed, paraffin-embedded human prostate carcinoma stained with peroxidase detection and DAB, but provides no comparison of fixation conditions (caption A02559-1). Compare sections processed together using the same pH 6.0, 95–98 °C, 20 min retrieval setting and identical detection timing (page retrieval setting; standard IHC practice). If staining still differs, inspect preservation and document fixation history where available before attributing the difference to TPI1 abundance (standard IHC practice).
How should I assess nuclear versus cytoplasmic TPI1 staining?
Score nuclear and cytoplasmic staining separately: UniProt places TPI1 in the cytoplasm, whereas HPA reports general nuclear and cytoplasmic tissue staining (UniProt P60174; HPA tissue IHC). HPA’s cell imaging lists the nucleoplasm as its approved main location, creating a compartment-specific discrepancy worth checking in the stained section (HPA subcellular). Use a counterstain and examine intact cells at high magnification so overlap from adjacent cytoplasm does not look intranuclear (standard IHC practice). Because HPA rates its tissue IHC reliability Uncertain, compare each compartment with controls before interpreting nuclear signal biologically (HPA tissue IHC; standard IHC practice).
Can this antibody distinguish TPI1 isoforms or modified epitopes in tissue?
The record lists 3 TPI1 isoforms and several modified residues, including phosphoserines at positions 21, 80, and 106 (UniProt P60174). The selected antibody is described as C-terminal in its tissue image, but the supplied evidence does not establish isoform coverage or modification-dependent binding (caption A02559-1; supplied evidence). Treat the chromogenic signal as antibody-reactive TPI1 unless epitope mapping or isoform-specific validation supports a narrower claim (standard IHC practice). When comparing conditions, keep retrieval and staining together across sections, since changes in epitope accessibility can complicate an apparent difference in abundance (standard IHC practice).
How can IF help resolve ambiguous TPI1 compartment staining?
Use IF as a separate validation experiment, pairing TPI1 with a marker for the expected cell type and a nuclear stain to assign signal to individual cells (standard IF practice). TPI1 lacks a transmembrane segment and is annotated in the cytoplasm, so use permeabilisation that allows antibody access to intracellular epitopes; the exact C-terminal epitope accessibility is unreported (UniProt P60174; caption A02559-1; standard IF practice). Choose a fluorophore channel away from the section’s strongest autofluorescence and include an unstained section to assess that background (standard IF practice). Compare nuclear and cytoplasmic signal independently because HPA cell imaging reports nucleoplasmic localisation while UniProt reports cytoplasm (HPA subcellular; UniProt P60174).
What should I check when TPI1 DAB staining looks diffuse?
Run a no-primary control through the same peroxidase detection and DAB development to reveal detection-related staining (standard IHC practice). Block endogenous peroxidase before detection, particularly when assessing signal in damaged or blood-containing areas, and compare those areas with well-preserved cells (standard IHC practice). The selected image demonstrates peroxidase-linked DAB staining in formalin-fixed, paraffin-embedded human prostate carcinoma, but does not establish the cause of diffuse signal in another section (caption A02559-1). If background persists, compare antibody dilution, blocking, wash conditions, and DAB development time on adjacent sections while checking whether cellular localisation remains interpretable (standard IHC practice).
How should I quantify TPI1 staining without conflating cell composition and intensity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and cell population before scoring, then report the percentage of positive cells and an intensity-weighted H-score for that population (standard IHC practice). Score nuclear and cytoplasmic compartments separately because both appear in HPA tissue IHC, while UniProt annotates cytoplasmic localisation (HPA tissue IHC; UniProt P60174). Normalise positive-cell counts to all evaluable cells of the same type, or report stained-cell density per mm² of viable tissue when area is the relevant denominator (standard IHC practice). Apply one threshold and development time across compared sections, and document exclusions for necrosis, folds, and poorly preserved edges (standard IHC practice).
When is a TPI1-positive cell credible rather than a staining artefact?
A credible result follows intact cell boundaries, appears in comparable well-preserved areas, and remains distinguishable from the no-primary control (standard IHC practice). Cytoplasmic signal fits UniProt’s localisation, while nuclear signal needs separate scrutiny because HPA reports it in tissue and cell imaging yet rates tissue IHC reliability Uncertain (UniProt P60174; HPA tissue IHC; HPA subcellular). Check whether the stained cell type is the one being scored; HPA reports medium staining in adipocytes and several glandular or epithelial populations, so mixed regions require cell-level review (HPA tissue IHC). Discount edge staining, necrotic deposits, and residual endogenous peroxidase signal before making a biological interpretation (standard IHC practice).
Boster reagents
Best TPI1 / Triosephosphate isomerase IHC Antibodies
One IHC-validated anti-TPI1 antibody has real staining data from formalin-fixed, paraffin-embedded human prostate carcinoma (A02559-1 figure caption). The catalog lists human, mouse and rat reactivity (A02559-1 catalog).
Real IHC data
Anti-TPI1 Antibody (C-term)
Cat # A02559-1
A02559-1 will render with a DAB IHC image of formalin-fixed, paraffin-embedded human prostate carcinoma (A02559-1 figure caption). Its listed applications include IHC-P and flow cytometry, and its listed reactivity covers human, mouse and rat (A02559-1 catalog).
Which to pick: Choose A02559-1 for paraffin-section tissue IHC: it is a rabbit polyclonal antibody with a listed IHC-P dilution of 1:50–1:100 (A02559-1 catalog), and its figure shows FFPE human prostate carcinoma (A02559-1 figure caption). No IF/ICC option is supported by this catalog payload because A02559-1 has no listed IF/ICC application or IF image (A02559-1 catalog). For mouse or rat IHC-P, A02559-1 lists reactivity with both species, although its supplied IHC figure shows human tissue only (A02559-1 catalog; A02559-1 figure caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P60174 (TPIS_HUMAN, Triosephosphate isomerase).
- Human Protein Atlas. TPI1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. TPI1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. TPI1 antibody validation summary (2 antibodies).
- Glycolysis- and immune-related novel prognostic biomarkers of Ewing's sarcoma: glucuronic acid epimerase and triosephosphate isomerase 1. Aging 2021 — PMC8312448.
- TPI1 Loss Triggers a Metabolite-Driven Mitochondrial Redox Vulnerability via the SARM1-cADPR-Ca(2+) Axis. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13379248.
- Identification of TPI1 As a potential therapeutic target in pancreatic cancer with dependency of TP53 mutation using multi-omics analysis. Cancer science 2024 — PMC11531968.
- Single-Cell RNA Sequencing Revealed That the Enrichment of TPI1(+) Malignant Hepatocytes Was Linked to HCC Metastasis and Immunosuppressive Microenvironment. Journal of hepatocellular carcinoma 2024 — PMC10896104.
- PubMed PMID:2579079 — UniProt-cited evidence.
- PubMed PMID:4022011 — UniProt-cited evidence.
- PubMed PMID:8723724 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TPI1 (Triosephosphate isomerase). Expected localisation Cytoplasmic expected (UniProt); nuclear and cytoplasmic observed (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02559-1 at 1:50-1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: FFPE human prostate carcinoma tissue (datasheet A02559-1); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A02559-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-TPI1, 1:50-1:100 (datasheet A02559-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Use formalin-fixed paraffin sections (selected-SKU IHC image A02559-1); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02559-1); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; C-terminal epitope coverage is unknown (UniProt; datasheet); Regulation: RNA tissue-enhanced in muscle, tongue (HPA tissue RNA). Evidence: UniProt P60174, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A02559-1, PMC11531968. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TPI1/immunohistochemistry-ihc-guide.