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TCP1 / T-complex protein 1 subunit alpha · IHC design guide
Design Immunohistochemistry for TCP1
Plan chromogenic IHC on paraffin sections around the cytoplasmic staining reported in most tissues (HPA tissue IHC). Use the reported high signal in gallbladder glandular cells and undetected signal in soft-tissue fibroblasts to inform control selection (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TCP1 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9826) | |
| Positive control | Gallbladder+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | No isoforms annotated; no signal peptide or TM segment (UniProt) |
Section 1
Recommended TCP1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet PB9826) is accompanied by published protocols for ovarian tissue (PMC8223286) and interspinous ligament samples (PMC8806538).
| Sample | Paraffin-embedded human ovary cancer tissue; fixative not specified (datasheet PB9826) |
| Fixation | Image fixative and duration unreported (datasheet PB9826); 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 PB9826); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9826) |
| Primary antibody | Rabbit anti-TCP1, 2-5μg/ml (datasheet PB9826) |
| Primary incubation | Overnight at 4 °C (datasheet PB9826) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9826) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TCP1-positive staining in glandular cells of gallbladder (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet PB9826). The interspinous ligament study used trypsin-EDTA retrieval (PMC8806538).
Section 2
What Is the Expected TCP1 Staining Pattern?
TCP1 is a cytosolic protein with a reported centrosome association and no transmembrane segment (UniProt P17987 localization and topology). In paraffin-section IHC, expect predominantly cytoplasmic staining across many tissues (HPA: cytoplasmic expression in most tissues). Gallbladder glandular cells, ovarian follicle cells and cells in testicular seminiferous ducts are reported as high-staining examples (HPA: High). HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA data (HPA: reliability).
| Cytoplasmic staining is clear in gallbladder glandular cells, ovarian follicle cells or cells in seminiferous ducts. | This matches the reported compartment and high-staining cell populations (HPA: cytoplasmic expression; High in these cells). Compare staining within identifiable cells and against a negative control on the same run (general IHC practice). A focal centrosomal accent may be compatible with UniProt localization, but the supplied HPA IHC record does not establish a required centrosomal pattern (UniProt P17987 localization; HPA: tissue IHC profile). |
| Strong nuclear-only or membrane-rim staining dominates while cytoplasm is clear. | Treat this as discordant with the expected cytoplasmic pattern and lack of a transmembrane segment (HPA: cytoplasmic expression; UniProt P17987 topology). Review morphology and the detection controls before assigning biological meaning (general IHC practice). The supplied sources do not validate nuclear-only or membrane-rim staining as a TCP1 IHC result. |
| Soft-tissue fibroblasts stain strongly, especially if expected glandular or follicle cells are weak. | HPA reports TCP1 as not detected in soft-tissue fibroblasts, while selected glandular and follicle cells stain at medium or high levels (HPA: tissue IHC). Unexpected fibroblast signal raises possible antibody cross-reactivity or endogenous detection activity; the image alone cannot distinguish them (general IHC practice). Compare with a no-primary control and an independently validated antibody (general IHC practice). |
| Chromogen spreads across extracellular spaces, tissue edges or many unrelated cells without clear cytoplasmic borders. | This distribution is difficult to score as cellular TCP1 because the reported tissue pattern is cytoplasmic (HPA: tissue IHC profile). Diffuse background can arise from nonspecific reagent binding or detection activity (general IHC practice). Check the no-primary control and staining distribution before calling weak positive cells (general IHC practice). |
| No cytoplasmic staining appears in a chosen high-staining reference tissue. | Absence in gallbladder glandular cells, ovarian follicle cells or cells in seminiferous ducts conflicts with their reported high levels (HPA: High in these populations). Check that the relevant cells are present, then review the run controls and assay settings (general IHC practice). This result alone does not show that TCP1 is absent from the specimen. |
💡Expected TCP1 appearanceCall a positive result when cytoplasmic chromogen is clearly cell-associated in the relevant cells, with strong examples in gallbladder glands, ovarian follicle cells or seminiferous-duct cells (HPA: cytoplasmic expression; High in these populations); nuclear-only, membrane-rim or diffuse extracellular signal is discordant with that pattern (HPA: tissue IHC profile; UniProt P17987 topology).
| Compartment and topology (UniProt P17987 localization and topology) | TCP1 is reported in cytosol and at the centrosome, with no transmembrane segment (UniProt P17987). Score the predominant cellular distribution against HPA's cytoplasmic IHC pattern; do not require visible centrosomal puncta in paraffin sections (HPA: tissue IHC profile; general IHC practice). |
| Cell-specific tissue distribution (HPA: tissue IHC) | High staining in selected gallbladder, ovary and testis cells contrasts with low staining in several listed cell populations and non-detection in soft-tissue fibroblasts (HPA: tissue IHC). Select a control by its documented cell population, then interpret weak signal in that cellular context. |
| Strength of tissue evidence (HPA: reliability and antibody records) | The tissue profile is Approved but has medium staining-to-RNA consistency; three listed antibodies have Approved IHC status (HPA: reliability; HPA: HPA027337, HPA031082, CAB017460). Those ratings support comparison with the reported pattern, without proving that every signal in a new specimen is specific. |
| IF/ICC evidence boundary (HPA: subcellular and antibody records) | HPA supplies no ICC/IF images or main IF location, and lists no ICC validation status for the three antibodies (HPA: subcellular record; HPA: antibody records). UniProt cytosolic and centrosomal localization can guide interpretation, but the supplied HPA record cannot establish an IF staining pattern (UniProt P17987 localization). |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-staining reference tissue gives no signal. | The expected positive cell population may be absent from the section, or the IHC run may have failed (HPA: High in selected cells; general IHC practice). | Confirm the named cell population on the section; review positive and no-primary controls, antigen retrieval, primary incubation and detection steps for the run (general IHC practice). Do not infer TCP1 absence from a failed control. |
| Signal is faint in glia, thyroid glands or cardiomyocytes. | Low staining is reported for these cell populations (HPA: Low in caudate glial cells, thyroid glandular cells and heart cardiomyocytes). | Compare with an HPA high-staining cell population processed in the same run, and score the weak signal only if it is cellular and above the negative control (HPA: High in selected populations; general IHC practice). |
| Staining is nuclear-only or outlines cell membranes. | That pattern conflicts with reported cytoplasmic IHC expression and the absence of a transmembrane segment (HPA: tissue IHC profile; UniProt P17987 topology). | Check the no-primary control and cell morphology; repeat with an independently IHC-approved antibody if the discordant pattern persists (general IHC practice; HPA: listed antibodies have Approved IHC status). |
| Fibroblasts stain strongly. | Soft-tissue fibroblasts are reported as not detected; cross-reactivity or endogenous detection activity is possible, but the cause is unresolved (HPA: soft-tissue fibroblasts; general IHC practice). | Compare a no-primary control and the expected positive cells on matched sections; if needed, compare an independently IHC-approved antibody (general IHC practice; HPA: antibody records). |
| Diffuse chromogen obscures cellular boundaries. | Nonspecific binding or endogenous detection activity may contribute background (general IHC practice); HPA reports a cytoplasmic cellular pattern (HPA: tissue IHC profile). | Inspect the no-primary control, verify blocking and detection steps, and adjust assay conditions using controls before scoring weak cytoplasmic staining (general IHC practice). |
| Can the IHC result be used as an IF/ICC pattern or protocol? | The supplied HPA record has no ICC/IF images, main IF location or ICC validation status for the listed antibodies (HPA: subcellular and antibody records). | Interpret IF/ICC separately using appropriate modality controls; use cytosolic and centrosomal localization only as a provisional expectation, not an HPA-validated IF pattern (UniProt P17987 localization; HPA: subcellular record; general IF practice). |
Experiment evidence
IHC & IF images showing TCP1 staining
PB9826 · IHCIHC analysis of TCP1 using anti-TCP1 antibody (PB9826). TCP1 was detected in a paraffin-embedded section of human ovary cancer tissue. Heat
PB9826 · IHCIHC analysis of TCP1 using anti-TCP1 antibody (PB9826). TCP1 was detected in a paraffin-embedded section of human testis tissue. Heat mediat
PB9826 · IFIF analysis of TCP1 using anti- TCP1 antibody (PB9077). TCP1 was detected in immunocytochemical section of CACO-2 cells. Enzyme antigen retr
M02389 · IHCIHC analysis of TCP1 alpha using anti-TCP1 alpha antibody (M02389). TCP1 alpha was detected in paraffin-embedded section of human lung cance
M02389 · IHCIHC analysis of TCP1 alpha using anti-TCP1 alpha antibody (M02389). TCP1 alpha was detected in paraffin-embedded section of human lung cance
M02389 · IFIF analysis of TCP1 alpha using anti-TCP1 alpha antibody (M02389). TCP1 alpha was detected in immunocytochemical section of MCF7 cells. Enzy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120438-TCP1/tissue/cerebral+cortex#imid_7000912
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120438-TCP1/tissue/colon#imid_7000838
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120438-TCP1/tissue/liver#imid_7000823
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000120438-TCP1/tissue/kidney#imid_7000824
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TCP1 IHC & IF
🧪Run ovary first and assess staining in follicle cells (HPA: High in ovary follicle cells); use soft tissue fibroblasts as the negative tissue (HPA: Not detected). On the ovary slide, treat other cell types as internal negatives only if their staining remains at background under matched controls; their TCP1 status is not established by the supplied HPA row (HPA: ovary follicle cells High).
Positive control tissue: Gallbladder (Glandular cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for TCP1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and clonality-matched rabbit IgG isotype controls, plus a TCP1 knockout specimen as a biological negative (caption: rabbit anti-TCP1; standard IHC practice). Check the ovary slide for endogenous peroxidase signal before interpreting DAB staining (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected PB9826 ovary cancer caption does not state a fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used in that paraffin-section example, but whether retrieval is required has not been established (caption: EDTA pH 8.0). The supplied evidence does not establish that frozen sections or IF are easier; for ovary, compare follicle-cell signal with background from the peroxidase control (HPA: ovary follicle cells High; caption: HRP/DAB detection).
HPA tissue IHC evidence for TCP1
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TCP1 IHC Tips
Troubleshoot TCP1 staining in paraffin sections by checking retrieval, compartment, tissue controls and detection conditions before interpreting signal.
How should I retrieve TCP1 when paraffin sections stain weakly?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet PB9826). The catalog antibody detected TCP1 in a paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet PB9826). If staining is weak, compare retrieval duration across matched sections while keeping antibody concentration and detection conditions fixed (standard IHC practice). Include a known positive section in each run, and compare cytoplasmic signal with tissue damage or diffuse background before increasing retrieval intensity (HPA: cytoplasmic expression in most tissues; standard IHC practice). The caption does not state the fixative, so this retrieval result does not establish performance across fixation conditions (datasheet PB9826).
Could fixation explain weak or uneven TCP1 staining?
TCP1-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (datasheet PB9826). Record the fixative and processing history for each specimen, then compare sections processed alike before attributing weak staining to TCP1 abundance (standard IHC practice). Keep heat retrieval in EDTA at pH 8.0 and the 2 μg/ml primary incubation constant during that comparison (datasheet PB9826). Inspect morphology and staining across the section, including edges, because uneven processing can complicate chromogenic interpretation (standard IHC practice). Neither TCP1's cytosolic location nor its annotated modified residues establishes a target-specific fixation effect (UniProt P17987: localisation and modified residues).
What staining pattern should I expect for TCP1 in tissue?
Expect predominantly cytoplasmic staining: TCP1 is annotated in the cytosol and at the microtubule organizing center or centrosome, while tissue IHC reports cytoplasmic expression in most tissues (UniProt P17987: subcellular location; HPA: tissue profile). Evaluate signal within identified cells rather than treating every stained area as equivalent (standard IHC practice). HPA reports high staining in ovary follicle cells and gallbladder glandular cells, but no detection in soft-tissue fibroblasts (HPA: tissue IHC). Compare those cell compartments with a counterstained section, and review isolated nuclear or extracellular deposits as possible artefacts before assigning them to TCP1 (HPA: tissue profile; standard IHC practice).
How do epitope accessibility and TCP1 variants affect staining?
The supplied record lists 0 annotated isoforms and no transmembrane segment; this does not identify the catalog antibody's epitope (UniProt P17987: isoforms and topology; datasheet PB9826). TCP1 has annotated modifications, including phosphoserine at residue 6 and phosphotyrosine at 181, but the payload does not link either site to antibody binding (UniProt P17987: modified residues; datasheet PB9826). If staining varies between specimens, check the antibody's epitope documentation and compare matched retrieval conditions before proposing a variant or modification effect (standard IHC practice). Interpret the observed signal as antibody-dependent TCP1 staining, with orthogonal confirmation if an epitope-specific biological conclusion is needed (standard IHC practice).
How can I investigate TCP1 by IF alongside tissue IHC?
For IF, pair TCP1 with a marker that identifies the cell population being examined, such as a marker for ovary follicle cells when studying that population (HPA: high in ovary follicle cells; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue autofluorescence before assigning a weak TCP1 channel (standard IF practice). Because TCP1 is cytosolic and lacks a transmembrane segment, use permeabilisation appropriate for access to an intracellular epitope, then assess preservation of cell morphology (UniProt P17987: location and topology; standard IF practice). The selected antibody evidence is a chromogenic paraffin-section result; it does not establish IF performance or an IF fixation condition (datasheet PB9826).
How do I reduce diffuse or misleading chromogenic background?
Compare a no-primary control with the TCP1-stained section to identify signal arising from detection reagents or tissue rather than primary-antibody binding (standard IHC practice). The selected paraffin-section procedure used 10% goat serum block, 2 μg/ml primary antibody overnight at 4°C, a peroxidase-conjugated secondary, and DAB development (datasheet PB9826). Check blocking, washes and primary-antibody concentration if staining spreads beyond cell boundaries (standard IHC practice). Include a peroxidase-blocking step when using chromogenic peroxidase detection, and compare any remaining deposit with the expected cytoplasmic distribution (standard IHC practice; HPA: tissue profile). Keep the counterstain light enough to distinguish cytoplasm from precipitate (standard IHC practice).
How should I score TCP1 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score cytoplasmic staining in a defined cell population and region, recording both the percentage of positive cells and intensity; an H-score can combine those observations (HPA: tissue profile; standard IHC practice). If cell counts are unreliable, report stained-cell density per mm² of viable tissue using the same threshold across sections (standard IHC practice). Normalise comparisons to the number of eligible cells or viable tissue area, and document exclusion of necrosis, folds and section edges (standard IHC practice). Keep retrieval at EDTA pH 8.0, detection settings and counterstaining consistent, because protocol differences can shift apparent intensity (datasheet PB9826; standard IHC practice).
When is TCP1 staining credible rather than artefactual?
A credible result follows the predominantly cytoplasmic pattern reported for TCP1 and is assessed in identified cells with preserved morphology (UniProt P17987: localisation; HPA: tissue profile; standard IHC practice). High signal in ovary follicle cells or gallbladder glandular cells is consistent with the HPA tissue observations, while staining assigned to soft-tissue fibroblasts warrants closer review (HPA: tissue IHC). Check for edge effects, necrosis and extracellular DAB deposits before scoring apparent positives (standard IHC practice). Compare a no-primary control and ensure peroxidase blocking is effective, because endogenous enzyme activity can imitate chromogenic signal (standard IHC practice). Treat unexpected nuclear-only staining as a finding requiring independent confirmation (UniProt P17987: localisation; standard IHC practice).
Boster reagents
Best TCP1 / T-complex protein 1 subunit alpha IHC Antibodies
IHC images show TCP1 staining in paraffin sections of human ovary cancer, lung cancer, placenta and testis, plus mouse and rat testis (PB9826 and M02389 IHC captions). IF is documented in MCF7 cells (M02389 IF caption).
Real IHC data
Anti-TCP1 alpha Antibody ®
Cat # PB9826
Real IHC data
Anti-TCP1 alpha Antibody ® (monoclonal, 2E7)
Cat # M02389
PB9826 will render with its human ovary cancer IHC figure; its other captions show human, mouse and rat testis paraffin sections (PB9826 IHC captions). M02389 will render with its human lung cancer IHC figure; its other captions show human placenta IHC and MCF7 cell IF (M02389 IHC and IF captions).
Which to pick: For paraffin section IHC across human, mouse and rat samples, choose rabbit PB9826 (PB9826 applications, reactivity and IHC captions); for human tissue IHC, mouse monoclonal M02389 also has paraffin section images (M02389 clone, reactivity and IHC captions). For IF/ICC, prefer M02389 because its own IF caption documents MCF7 cells; PB9826 lists IF/ICC, but its supplied IF caption names PB9077 (M02389 IF caption; PB9826 applications and IF caption). Both IHC captions omit the fixative, so fixation conditions are unreported (PB9826 and M02389 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P17987 (TCPA_HUMAN, T-complex protein 1 subunit alpha).
- Human Protein Atlas. TCP1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TCP1 subcellular location (ICC-IF): Highest expression in HAP1: 728.0 nTPM.
- Human Protein Atlas. TCP1 antibody validation summary (3 antibodies).
- TCP1 expression alters the ferroptosis sensitivity of diffuse large B-cell lymphoma subtypes by stabilising ACSL4 and influences patient prognosis. Cell death & disease 2024 — PMC11341815.
- TCP1 regulates PI3K/AKT/mTOR signaling pathway to promote proliferation of ovarian cancer cells. Journal of ovarian research 2021 — PMC8223286.
- Low expression of TCP1 (T-Complex 1) and PSMC1 (Proteasome 26S subunit, ATPase 1) in heterotopic ossification during ankylosing spondylitis. Bioengineered 2021 — PMC8806538.
- TCP1 promotes the progression of malignant tumours by stabilizing c-Myc through the AKT/GSK-3β and ERK signalling pathways. Communications biology 2025 — PMC11971430.
- PubMed PMID:2377466 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TCP1 (T-complex protein 1 subunit alpha). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9826). Validated IHC antibody: PB9826 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovary cancer tissue; fixative not specified (datasheet PB9826); 2. Fixation: Image fixative and duration unreported (datasheet PB9826); 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 PB9826); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9826); 8. Primary antibody: Rabbit anti-TCP1, 2-5μg/ml (datasheet PB9826); 9. Primary incubation: Overnight at 4 °C (datasheet PB9826). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms annotated; no signal peptide or TM segment (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt P17987, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in HAP1: 728.0 nTPM, Datasheet PB9826, PMC8223286, PMC8806538. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TCP1/immunohistochemistry-ihc-guide.