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TPBG / Trophoblast glycoprotein · IHC design guide
Design Immunohistochemistry for TPBG
Use this guide to plan TPBG chromogenic IHC on paraffin sections, with kidney tubules as a high-staining control and hippocampal glia as a no-signal comparison (HPA tissue IHC). Assess cytoplasmic staining with a membranous pattern (HPA tissue IHC) alongside TPBG’s cell membrane topology (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TPBG 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 with a membranous pattern (HPA tissue IHC) | |
| Staining pattern | Glandular cells: cytoplasmic with a membranous pattern (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Hippocampus+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Signal varies by cell type; glia and chondrocytes are undetected (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms listed; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended TPBG IHC & IF Protocols
The catalog antibody protocol is paired with one published TPBG protocol for chromogenic IHC on paraffin sections (PMC12208316).
| Sample | Paraffin-embedded human uters cancer tissue; fixative not specified (datasheet M07442) |
| Fixation | Image fixative and duration unreported (datasheet M07442); 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 monoclonal (clone EHE-20) anti-TPBG, 1:50 (datasheet M07442) |
| 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 | TPBG-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with a membranous pattern in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the published protocol also uses citrate pH 6.0 (PMC12208316).
Section 2
What Is the Expected TPBG Staining Pattern?
TPBG is a cell-membrane protein with an extracellular region spanning residues 32–355 and a transmembrane segment at 356–376 (UniProt Q13641 topology). In paraffin-section IHC, expect cytoplasmic staining with a membranous pattern in selected cells (HPA tissue IHC: Enhanced reliability). HPA reports high staining in kidney tubule cells, lung type I alveolar cells, and several glandular cell populations; expression varies by cell type (HPA tissue IHC).
| Cell edges are outlined, with accompanying cytoplasmic stain in the expected cells. | This fits the reported tissue IHC pattern (HPA: cytoplasmic expression with a membranous pattern). Membrane-associated signal is consistent with TPBG topology (UniProt Q13641: cell membrane; extracellular residues 32–355). Score the identified cell population, because an adjacent unstained population does not negate a positive result. |
| Kidney tubule cells or lung type I alveolar cells stain strongly, while neighboring cells differ. | These are useful positive-reference populations (HPA: High in kidney tubule cells; High in lung type I alveolar cells). Compare staining within the named population and against local background. HPA describes tissue IHC staining, so a strong result in one population does not establish that every cell in that tissue must stain. |
| Staining is exclusively nuclear, without a convincing cytoplasmic or membranous IHC pattern. | Investigate the compartment assignment: nuclear-only staining differs from the tissue IHC profile (HPA: cytoplasmic expression with a membranous pattern). Interpret cautiously because the separate ICC-IF record reports nucleoplasm as an approved location, while this antibody's ICC validation is Uncertain (HPA subcellular; HPA antibody HPA010554). |
| Unexpected cells stain, including glial cells in hippocampus or chondrocytes in soft tissue. | HPA reports TPBG as Not detected in those specific cell populations (HPA tissue IHC). Recheck cell identification and compare with controls; unexpected chromogen could reflect cross-reactivity or endogenous detection activity (general IHC practice). A result in those cells warrants investigation, not an automatic conclusion about the whole tissue. |
| The section shows broad haze, or a known high-staining population has no signal. | Diffuse staining that obscures cell boundaries does not establish TPBG localisation; assess background controls (general IHC practice). Absence of signal in a high-staining reference population calls for a run check before interpreting the study section as negative (HPA: High in kidney tubule cells and other listed populations). |
💡Expected TPBG appearanceCall positive when the named cell population shows discernible cytoplasmic staining with a membranous pattern above background—potentially strong in HPA High populations; diffuse haze or unexplained staining in HPA Not detected cell populations is suspect (HPA tissue IHC).
| Cell population and tissue | HPA grades selected glandular cells, kidney tubule cells, lung type I alveolar cells, and skin eccrine glands High; other populations are Low or Not detected (HPA tissue IHC). Choose and score controls by cell population rather than assuming uniform staining across a section. |
| Protein topology | TPBG has an extracellular region at 32–355, a transmembrane segment at 356–376, and a cytoplasmic tail at 377–420 (UniProt Q13641 topology). This supports a membrane-associated interpretation, but no antibody epitope is supplied; topology alone cannot predict retrieval requirements. |
| Evidence strength and antibody scope | The tissue IHC profile has Enhanced reliability, described as medium consistency with RNA data (HPA tissue IHC). Antibody HPA010554 is IHC Enhanced but ICC Uncertain (HPA antibodies). These labels support use of the tissue IHC pattern while preserving uncertainty about results in a different application. |
| IF/ICC Q&A: should nuclear IF determine the IHC score? | No. The separate ICC-IF record lists nucleoplasm as an approved location (HPA subcellular), whereas tissue IHC reports a cytoplasmic pattern with membranous staining (HPA tissue IHC). The listed antibody's ICC status is Uncertain (HPA antibodies); resolve an IHC score using the IHC slide and controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a reference population expected to stain strongly. | A run-level detection or tissue problem is possible; the result alone cannot identify its cause (general IHC practice). | Check section integrity, primary-antibody addition, retrieval and detection records, then compare with a concurrently stained positive control. Kidney tubule cells are one documented High population (HPA tissue IHC). |
| Cytoplasmic colour is present, but cell borders cannot be distinguished. | High background, excess detection signal, or poor morphological resolution can obscure the reported pattern (general IHC practice; HPA tissue IHC: cytoplasmic with membranous pattern). | Review a negative control and tissue morphology; adjust general IHC blocking, washing, antibody concentration, or detection development as the control findings indicate (general IHC practice). |
| Colour appears broadly across the section, including cell-free areas. | Nonspecific reagent deposition or endogenous detection activity may contribute (general IHC practice). | Inspect no-primary and detection controls; address the implicated detection chemistry and washing before scoring TPBG (general IHC practice). |
| Only nuclei stain in an IHC section. | The compartment differs from the reported tissue IHC pattern; the cause cannot be determined from this appearance alone (HPA tissue IHC). | Check the counterstain and detection controls, then reassess localisation at adequate magnification (general IHC practice). Keep the ICC-IF nucleoplasm report separate because ICC validation is Uncertain (HPA subcellular; HPA antibodies). |
| Unexpected cell populations stain more strongly than the intended population. | Cell misidentification, cross-reactivity, or endogenous activity are possibilities (general IHC practice). HPA grades hippocampal glial cells and soft-tissue chondrocytes Not detected (HPA tissue IHC). | Confirm cell identity from morphology, compare with the HPA cell-specific profile, and inspect negative controls before treating the signal as TPBG (HPA tissue IHC; general IHC practice). |
| A study section appears negative while the control stains appropriately. | TPBG staining differs across cell populations (HPA tissue IHC), and an appropriate run control does not establish expression in the study population (general IHC practice). | Record the specific cells assessed and their background; report absence of detectable IHC staining in that population under these conditions. Avoid extending that finding to other cells or tissues (general IHC practice). |
Experiment evidence
IHC & IF images showing TPBG staining
M07442 · IHCImmunohistochemical analysis of paraffin-embedded human uters cancer, using 5T4 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000146242-TPBG/tissue/cerebral+cortex#imid_2926824
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000146242-TPBG/tissue/colon#imid_2926750
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000146242-TPBG/tissue/liver#imid_2926767
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000146242-TPBG/tissue/kidney#imid_2926760
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA010554; green = TPBG, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA010554; green = TPBG, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TPBG IHC & IF
🧪Run kidney first: tubular cells should stain (HPA: High in cells in tubules). Use hippocampus glial cells as the negative comparison (HPA: Not detected in glial cells); on the kidney slide, surrounding nontubular cells should show only background signal, with their TPBG status verified independently.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Hippocampus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TPBG in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, subclass and concentration; and TPBG knockout tissue where available (standard IHC practice). For kidney, quench endogenous peroxidase and, if using avidin–biotin detection, block endogenous biotin or use polymer detection to assess tubular background (standard IHC practice).
⚠️Feasibility: No TPBG-specific fixation window or antigen-retrieval effect is reported in the supplied evidence; the fixative is unreported in the selected M07442 paraffin-section caption (caption: paraffin-embedded tissue, fixative not stated). Optimize retrieval on matched paraffin sections; the evidence does not establish that frozen sections or IF/ICC are easier (caption: paraffin-embedded tissue; HPA: ICC-IF images). Kidney tubular endogenous biotin can create staining artefacts with avidin–biotin detection (standard IHC practice).
HPA tissue IHC evidence for TPBG
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type I | High | Protein (IHC) | HPA → |
Section 3
Advanced TPBG IHC Tips
Use the documented paraffin section image and TPBG localisation data to plan controls, then optimise staining and scoring within each specimen series.
Where should I start antigen retrieval for TPBG in paraffin sections?
Start with citrate pH 6.0 heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). The M07442 image shows staining in paraffin-embedded human uters cancer, but its caption gives neither a fixative nor a retrieval method (M07442 caption). Compare a retrieved section with a matched section processed without retrieval, keeping detection and imaging conditions constant (standard IHC practice). If staining remains weak, test Tris-EDTA pH 9 as a fallback on adjacent sections and check whether tissue integrity and background deteriorate (standard IHC practice). Score the cell surface and cytoplasmic patterns separately because both occur in tissue staining (UniProt Q13641 localisation; HPA tissue IHC profile).
How should I handle fixation when TPBG staining varies between blocks?
Target-specific fixation sensitivity is unknown: the M07442 caption identifies paraffin-embedded tissue but does not report its fixative or fixation duration (M07442 caption). For new specimens, record the fixative, time to fixation, and duration, then process comparison sections together using a consistent 10% neutral buffered formalin workflow (standard IHC practice). Compare blocks with similar tissue preservation before changing the antibody concentration or the page’s citrate pH 6.0 retrieval setting (page retrieval setting; standard IHC practice). Include an internal positive cell population where the specimen provides one, and interpret absent staining cautiously when preanalytic histories differ (HPA tissue IHC profile; standard IHC practice). Neither tissue staining patterns nor protein topology establishes how this target responds to fixation (HPA tissue IHC profile; UniProt Q13641 topology).
Should TPBG staining be membranous, cytoplasmic, or nuclear?
Prioritise cell surface staining when assessing the proposed membrane pool: TPBG has an extracellular region at residues 32–355 and a transmembrane segment at 356–376 (UniProt Q13641 topology). Tissue IHC also reports cytoplasmic expression with a membranous pattern, so record these compartments separately (HPA tissue IHC profile). The subcellular IF record lists approved nucleoplasmic localisation, which merits a separate nuclear score and orthogonal verification rather than automatic reassignment to the membrane signal (HPA subcellular record). On adjacent sections, compare the suspect pattern with a no-primary control and with cell morphology under the counterstain (standard IHC practice). Document whether staining follows intact cell borders, diffuse cytoplasm, or nuclei before drawing a biological conclusion (standard IHC practice).
Could epitope position or glycosylation explain discordant TPBG staining?
First establish the antibody’s mapped immunogen or epitope, if available; no epitope position is supplied for M07442 (M07442 caption). TPBG exposes residues 32–355 outside the cell, spans the membrane at 356–376, and has a cytoplasmic tail at 377–420 (UniProt Q13641 topology). Its annotated glycosylation sites are residues 81, 124, and 275, making epitope accessibility a reasonable hypothesis to test rather than an established cause of weak staining (UniProt Q13641 glycosylation). No isoforms are annotated in the supplied record, so do not assign differing compartments to a named isoform (UniProt Q13641 isoform record). Compare retrieval conditions on adjacent sections while preserving identical detection and scoring criteria (standard IHC practice).
How can IF help resolve ambiguous TPBG localisation in stained tissue?
On a matched IF section, multiplex TPBG with a marker for the expected cell type, such as a trophoblast lineage marker in trophoblast tissue, and inspect overlap cell by cell (UniProt Q13641 tissue specificity; standard IF practice). Choose a red or far-red fluorophore after checking tissue autofluorescence in an unstained section, and keep exposure settings fixed across controls (standard IF practice). For an extracellular epitope within residues 32–355, compare staining without permeabilisation; a cytoplasmic-tail epitope within 377–420 requires access through the membrane (UniProt Q13641 topology; standard IF practice). Because the M07442 epitope is unspecified, confirm its location before interpreting that comparison (M07442 caption). Evaluate nuclear signal separately because the IF subcellular record reports nucleoplasmic localisation (HPA subcellular record).
How do I distinguish TPBG signal from chromogenic background?
Inspect a no-primary section alongside each run to identify colour produced by tissue or detection reagents without the primary antibody (standard IHC practice). For peroxidase detection, test a 3% hydrogen peroxide block for 10 min and verify that residual endogenous activity does not resemble cellular DAB staining (standard IHC practice). If diffuse colour persists, optimise blocking, washing, and antibody concentration on serial sections while holding retrieval at citrate pH 6.0 constant (page retrieval setting; standard IHC practice). Compare staining within preserved cell boundaries against the expected membrane and cytoplasmic tissue patterns, especially near folds or section edges (UniProt Q13641 localisation; HPA tissue IHC profile; standard IHC practice). Record pigment and precipitate separately from scored cells under the counterstain (standard IHC practice).
What scoring method captures heterogeneous TPBG staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define membrane and cytoplasmic compartments before scoring because TPBG is annotated at the cell membrane while tissue IHC also reports cytoplasmic staining (UniProt Q13641 localisation; HPA tissue IHC profile). For each compartment, report the percentage of viable target cells at intensity 0–3, or calculate an H-score as the sum of intensity times percentage, spanning 0–300 (standard IHC practice). Normalise positive counts to the number of viable cells in the defined cell population, or report positive-cell density per mm² of viable tissue (standard IHC practice). Exclude necrosis, folds, and detached edges by a prespecified rule, then use the same thresholds and sampling plan for every section (standard IHC practice). Keep nuclear scores separate if investigating the reported IF localisation (HPA subcellular record).
When should I doubt an apparent TPBG-positive result?
Check whether staining belongs to the intended cells: TPBG is reported in trophoblasts and several carcinoma types, while tissue IHC also identifies positive glandular and other defined cell populations (UniProt Q13641 tissue specificity; HPA tissue IHC profile). A rim restricted to section edges, colour over necrosis, or staining that survives a no-primary control warrants investigation before scoring (standard IHC practice). Membrane and cytoplasmic signals fit the tissue IHC profile, whereas a nuclear-only chromogenic result needs independent confirmation because the IF subcellular record reports nucleoplasmic localisation (HPA tissue IHC profile; HPA subcellular record). For peroxidase detection, verify the 3% hydrogen peroxide block when endogenous enzyme could produce DAB colour (standard IHC practice). Interpret weak or absent staining against matched positive tissue and processing controls, not morphology alone (standard IHC practice).
Boster reagents
Best TPBG / Trophoblast glycoprotein IHC Antibodies
The catalog antibody has a human paraffin-section IHC image (M07442 image caption; catalog: Human, IHC). No IF image or IF application is listed (catalog: applications and image captions).
Real IHC data
Anti-5T4 TPBG Rabbit Monoclonal Antibody
Cat # M07442
M07442 will render with an IHC image captioned as paraffin-embedded human “uters cancer” (M07442 image caption). Its listed reactivity is Human and its applications include IHC, with no IF application listed (catalog: M07442 applications and reactivity).
Which to pick: Choose M07442 for human tissue IHC: it is a rabbit monoclonal with a paraffin-section IHC image (catalog: M07442 host and clone; M07442 image caption). The caption does not report the fixative (M07442 image caption). No SKU in the payload is listed for IF/ICC or cross-species use (catalog: M07442 applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13641 (TPBG_HUMAN, Trophoblast glycoprotein).
- Human Protein Atlas. TPBG tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. TPBG subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. TPBG antibody validation summary (1 antibodies).
- Trophoblast glycoprotein is a marker for efficient sorting of ventral mesencephalic dopaminergic precursors derived from human pluripotent stem cells. NPJ Parkinson's disease 2021 — PMC8289854.
- Proteogenomic analysis of the CALGB 40601 (Alliance) HER2+ breast cancer neoadjuvant trial reveals resistance biomarkers. Cell reports. Medicine 2025 — PMC12208316.
- Expression of Oncofetal Antigen 5T4 in Murine Taste Papillae. Frontiers in cellular neuroscience 2019 — PMC6685444.
- Combining data from TCGA and GEO databases and reverse transcription quantitative PCR validation to identify gene prognostic markers in lung cancer. OncoTargets and therapy 2019 — PMC6345189.
- PubMed PMID:8132670 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16303743 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TPBG (Trophoblast glycoprotein). Expected localisation Cytoplasmic staining with a membranous pattern (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M07442 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Hippocampus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human uters cancer tissue; fixative not specified (datasheet M07442); 2. Fixation: Image fixative and duration unreported (datasheet M07442); 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 monoclonal (clone EHE-20) anti-TPBG, 1:50 (datasheet M07442); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Signal varies by cell type; glia and chondrocytes are undetected (HPA tissue IHC); Isoform / epitope: No isoforms listed; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q13641, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet M07442, PMC12208316. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TPBG/immunohistochemistry-ihc-guide.