This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TNFRSF10B / Tumor necrosis factor receptor superfamily member 10B · IHC design guide
Design Immunohistochemistry for TNFRSF10B
Plan TNFRSF10B IHC in paraffin sections using cytoplasmic tissue staining as the reference pattern (HPA tissue IHC). This guide covers fixation, controls and interpretation of extracellular versus cytoplasmic epitopes based on receptor topology (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TNFRSF10B 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 tissue staining (HPA tissue IHC); membrane topology (UniProt) | |
| Staining pattern | Cytoplasmic staining across many tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00410) | |
| 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 | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may differ from RNA expression (HPA tissue IHC) | |
| Regulation | HCMV UL141 blocks surface expression (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope side matters; coverage unknown (UniProt) |
Section 1
Recommended TNFRSF10B IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA retrieval (datasheet A00410). The published IHC protocols below cover synovial tissue (PMC13607894 methods) and lung autopsy sections (PMC10417228 methods).
| Sample | Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A00410) |
| Fixation | Image fixative and duration unreported (datasheet A00410); 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 A00410); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00410) |
| Primary antibody | Rabbit anti-TNFRSF10B, 0.5-1μg/ml (datasheet A00410) |
| Primary incubation | Overnight at 4 °C (datasheet A00410) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00410) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TNFRSF10B-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA, pH 8.0 (datasheet A00410); for lung autopsy sections, compare citrate, pH 6.0, at 105°C (PMC10417228 methods).
Section 2
What Is the Expected TNFRSF10B Staining Pattern?
TNFRSF10B is a membrane receptor with an extracellular region at residues 56–210 and a cytoplasmic region at 232–440 (UniProt O14763 topology). On paraffin sections, expect broad cellular staining, often cytoplasmic, with strong staining in several listed cell populations (HPA: ubiquitous cytoplasmic expression; High in listed cells). Interpret intensity with care: HPA rates tissue IHC reliability Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Adipocytes, bronchial respiratory epithelium, or breast glandular cells show strong cellular chromogen, sometimes outlining cells. | High staining in these cell populations is consistent with HPA tissue IHC (HPA: High in adipocytes, respiratory epithelial cells, and breast glandular cells). A resolved cell-edge pattern is compatible with membrane localisation (UniProt O14763 topology; HPA subcellular: Membrane). Record cytoplasmic and edge staining separately: HPA describes the overall tissue pattern as cytoplasmic (HPA tissue IHC). |
| Cytoplasmic staining is widespread, but crisp membrane outlines are absent. | This can match the reported tissue pattern; it does not, by itself, establish surface receptor staining (HPA tissue IHC: ubiquitous cytoplasmic expression; UniProt O14763: Membrane). Assess whether signal follows cell boundaries in a well-preserved positive area. The HPA subcellular summary says Membrane but supplies no ICC-IF images here (HPA subcellular). |
| Only nuclei stain, or the deposit follows folds, debris, or section edges. | A nuclear-only pattern conflicts with the membrane annotation and reported cytoplasmic tissue pattern (UniProt O14763: Membrane; HPA tissue IHC). Treat deposit confined to tissue damage as suspect in routine chromogenic IHC; compare a clean area and the negative detection control before scoring it as TNFRSF10B. |
| The strongest signal falls in cells expected to stain weakly, such as hippocampal glia, while listed high-staining cells remain faint. | HPA reports low staining in hippocampal glia and high staining in selected cell populations (HPA tissue IHC). Check cell identification and staining controls before calling this a true reversal. Cross-reactivity or endogenous detection activity is possible in IHC; distribution alone cannot distinguish them. |
| A known HPA high-staining cell population has no visible signal. | An absent result in adipocytes, bone-marrow hematopoietic cells, or colon endothelial cells warrants a control review (HPA: High in these cells). Confirm that those cells are present, then review the catalog antibody's IHC-P instructions, antigen retrieval, and detection controls. HPA's Approved rating has medium RNA–staining consistency, so one negative section does not settle target absence (HPA tissue IHC). |
💡Expected TNFRSF10B appearanceCall a section positive when identifiable HPA high-staining cells show convincing cellular chromogen, often cytoplasmic and potentially membrane-associated; nuclear-only or damage-associated deposit is suspect (HPA tissue IHC; UniProt O14763 topology).
| Compartment and epitope orientation | The receptor spans residues 211–231, with extracellular and cytoplasmic regions on either side (UniProt O14763 topology). Interpret a cell-edge signal in that context. The supplied antibody record does not identify its epitope (HPA antibody record: HPA023625), so topology cannot predict which region its stain reports. |
| Tissue pattern versus receptor annotation | HPA calls the tissue IHC pattern ubiquitous and cytoplasmic, while UniProt and HPA subcellular annotate membrane localisation (HPA tissue IHC; HPA subcellular; UniProt O14763). Preserve that distinction when scoring. Cytoplasmic chromogen alone cannot show whether receptor is present at the cell surface. |
| Tissue and cell selection | Use the named cell population as the comparison unit: HPA lists high staining in adipocytes, marrow hematopoietic cells, and colon endothelial cells, but low staining in hippocampal and caudate glia (HPA tissue IHC). An entire tissue should not receive one intensity score when its cell populations differ. |
| Source and antibody certainty | HPA marks the tissue IHC profile Approved, with medium consistency against RNA data, and records HPA023625 as IHC Approved (HPA tissue IHC; HPA antibodies). These ratings support comparison with the displayed pattern but do not establish specificity in every specimen. The supplied record has no target-specific fixation-sensitivity result. |
| Isoforms and processing | UniProt lists 3 isoforms and a signal peptide at residues 1–55; the annotated mature chain spans 56–440 (UniProt O14763). Because the supplied antibody record gives no epitope, a particular isoform contribution or epitope loss cannot be inferred from an IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA high-staining cells are blank. | The expected cells may be absent from the section, or an IHC workflow step may have failed; a negative slide alone does not identify the cause (HPA: High in the named cells). | Locate the named cells on the counterstain, include a separately processed positive tissue, and verify retrieval, primary-antibody incubation, and chromogenic detection against the catalog antibody's IHC-P instructions. |
| Chromogen covers cells and stroma uniformly. | Diffuse deposit can reflect background from detection chemistry or insufficient blocking in routine IHC; it is hard to reconcile with a cell-resolved pattern (HPA tissue IHC: cellular staining profile). | Compare a primary-omission control, inspect reagent and wash steps, and adjust blocking or detection conditions using the catalog IHC-P procedure. Score cellular signal only after the diffuse background is resolved. |
| Nuclear staining dominates. | This conflicts with membrane localisation and HPA's reported cytoplasmic tissue pattern (UniProt O14763; HPA tissue IHC). A nuclear deposit should therefore be treated as an unverified IHC signal. | Inspect morphology and a primary-omission control; compare nuclear staining with the same run's positive tissue. Reassess primary-antibody and detection conditions before assigning a TNFRSF10B score. |
| Unexpected cells stain more strongly than the anticipated cells. | Cell misidentification, nonspecific binding, or endogenous chromogen-generating activity may explain the mismatch in routine IHC; HPA assigns different levels to specific cells (HPA tissue IHC). | Identify cells by morphology and counterstain, compare the HPA cell-level pattern, and run an appropriate negative detection control. For enzyme detection, check the applicable endogenous-activity blocking step. |
| A slide is called negative because membrane outlines are faint. | HPA describes ubiquitous cytoplasmic tissue staining despite membrane localisation annotations (HPA tissue IHC; HPA subcellular; UniProt O14763). Requiring crisp outlines could discard a reported pattern. | Score reproducible cellular cytoplasmic staining separately from membrane-associated staining. Use the same compartment criteria in controls and study sections; do not infer surface receptor abundance from cytoplasmic chromogen alone. |
| Cerebellar Purkinje cells stain despite an expectation of no brain signal. | The sources differ: HPA lists high Purkinje-cell IHC staining, whereas UniProt reports TNFRSF10B as undetectable in brain in its tissue-specificity summary (HPA tissue IHC; UniProt O14763 tissue specificity). | Document the cell-specific observation and the source disagreement. Review staining controls and antibody specificity before treating the Purkinje-cell signal as confirmed TNFRSF10B or rejecting it solely from the UniProt summary. |
Experiment evidence
IHC & IF images showing TNFRSF10B staining
A00410 · IHCIHC analysis of DR5 using anti-DR5 antibody (A00410). DR5 was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated an
A00410 · IHCIHC analysis of DR5 using anti-DR5 antibody (A00410). DR5 was detected in a paraffin-embedded section of mouse intestine tissue. Heat mediat
A00410-2 · IHCImmunohistochemistry Validation of DR5 in Mouse kidney tissue Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using an
A00410-2 · IHCImmunohistochemistry Validation of BIM in Human Colon Tumors (Devetzi et al., 2016) Protein analysis for DR5 by immunohistochemistry with an
A00410-2 · IFImmunofluorescence Validation of DR5 in Human HepG2 Cells Immunofluorescent analysis of 4% paraformaldehyde-fixed human HepG2 cells labeling
A00410-5 · IHCIHC analysis of DR5/TNFRSF10B using anti-DR5/TNFRSF10B antibody (A00410-5). DR5/TNFRSF10B was detected in a paraffin-embedded section of hum
A00410-5 · IHCIHC analysis of DR5/TNFRSF10B using anti-DR5/TNFRSF10B antibody (A00410-5). DR5/TNFRSF10B was detected in a paraffin-embedded section of hum
PA1842 · IHCAnti-DR5 antibody, PA1842, IHC(P) IHC(P): Human Lung Cancer Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000120889-TNFRSF10B/tissue/cerebral+cortex#imid_6011016
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000120889-TNFRSF10B/tissue/colon#imid_6011534
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000120889-TNFRSF10B/tissue/liver#imid_6011535
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000120889-TNFRSF10B/tissue/kidney#imid_6011528
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TNFRSF10B IHC & IF
🧪Run breast first and score glandular cells for staining (HPA: High in breast glandular cells). HPA detects TNFRSF10B in all 45 scored tissues, so use no-primary and isotype controls for negative comparison; cells without specific signal on the positive slide should show only counterstain, but no named cell population is established as an internal negative (HPA: no negative tissue rows).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: TNFRSF10B 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 carries no ICC-IF cell line for TNFRSF10B; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only slide, a concentration-matched rabbit IgG isotype control, and TNFRSF10B knockout tissue or cells as a biological negative (selected A00410 caption: rabbit primary antibody). For DAB detection, quench endogenous peroxidase and control endogenous biotin because the selected method uses a streptavidin–biotin complex (selected A00410 caption: SABC with DAB).
⚠️Feasibility: The selected paraffin-section caption does not report a fixative, and the supplied evidence reports no target-specific fixation window or fixation effect (selected A00410 caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 precedes staining at 1 μg/ml overnight at 4°C, establishing a starting IHC condition without establishing retrieval dependence (selected A00410 caption). There is no supplied comparison showing that frozen sections or IF are easier; the membrane location supports checking membrane-associated signal, while adipose tissue may require care in interpreting sparse adipocyte cytoplasm if chosen for follow-up (UniProt O14763: membrane; HPA: High in adipocytes).
HPA tissue IHC evidence for TNFRSF10B
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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: TNFRSF10B is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced TNFRSF10B IHC Tips
Troubleshoot TNFRSF10B staining by checking retrieval, antibody access, compartmental pattern and controls before comparing signal across sections.
What retrieval conditions should I start with for TNFRSF10B paraffin sections?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet A00410). The catalog antibody produced a rat kidney IHC image after this retrieval, followed by 1 μg/ml primary antibody overnight at 4°C (datasheet A00410). If staining is weak, compare retrieval duration on adjacent sections while holding antibody concentration, detection and development constant; excessive heating can damage morphology (general IHC practice). Include a no-primary control and inspect intact tissue away from section edges, since stronger DAB alone does not establish specific TNFRSF10B staining (general IHC practice).
How should I troubleshoot fixation when TNFRSF10B staining is weak?
The A00410 image shows a paraffin-embedded section, but its fixative is unreported; target-specific fixation sensitivity is therefore unknown (datasheet A00410). Record fixation and processing history before comparing sections, and process matched specimens together where possible (general IHC practice). Start troubleshooting with the documented EDTA pH 8.0 retrieval and 1 μg/ml primary antibody overnight at 4°C (datasheet A00410). If matched sections still differ, assess tissue preservation and vary one processing or retrieval condition at a time, using a no-primary control to check whether the difference reflects nonspecific detection (general IHC practice).
Should TNFRSF10B appear at the membrane or in the cytoplasm?
TNFRSF10B is annotated as a membrane protein with an extracellular region at residues 56–210, a transmembrane segment at 211–231 and a cytoplasmic region at 232–440 (UniProt O14763 topology). HPA describes its tissue IHC pattern as ubiquitous cytoplasmic expression, with medium consistency between staining and RNA data (HPA tissue IHC). In chromogenic sections, score membrane-associated and cytoplasmic staining separately rather than assigning every brown area to surface receptor (general IHC practice; UniProt O14763 topology). Compare the same cell types across matched sections and use a no-primary control to evaluate diffuse cytoplasmic DAB before calling it TNFRSF10B (general IHC practice).
Could isoform choice or epitope location explain uneven TNFRSF10B staining?
UniProt lists three TNFRSF10B isoforms—Long, 3 and Short—but the supplied IHC caption does not identify the antibody epitope (UniProt O14763 isoforms; datasheet A00410). The annotated extracellular region spans residues 56–210, while residues 232–440 are cytoplasmic (UniProt O14763 topology). Check the antibody's documented immunogen or epitope before inferring that a negative section lacks every isoform; the supplied evidence cannot establish isoform coverage (UniProt O14763 isoforms; datasheet A00410). When comparing staining patterns, keep retrieval and detection matched, and report the antibody identifier alongside the scored compartment (general IHC practice).
How can IF help resolve ambiguous TNFRSF10B IHC localisation?
Use IF as a separate localisation check, since the supplied A00410 example documents chromogenic paraffin IHC and supplies no IF validation (datasheet A00410). Multiplex TNFRSF10B with a marker for the cell type being examined, choosing a spectrally separated fluorophore in a channel with low tissue autofluorescence (general IF practice). Because TNFRSF10B has extracellular and cytoplasmic regions, determine the antibody epitope before deciding whether permeabilisation is needed to expose the cytoplasmic side (UniProt O14763 topology; general IF practice). Include single-label and no-primary controls, then compare IF localisation with membrane-associated and cytoplasmic IHC scores without treating either signal alone as proof of surface expression (general IF/IHC practice).
How do I reduce diffuse DAB background without losing TNFRSF10B signal?
First inspect a no-primary section for detection background and compare it with the full stain at identical DAB development time (general IHC practice). The A00410 example used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, and biotinylated secondary with SABC detection (datasheet A00410). For diffuse signal, check wash consistency and titrate primary antibody around the documented concentration while keeping retrieval fixed (general IHC practice; datasheet A00410). With biotin-based detection, consider endogenous biotin controls; use a peroxidase block and review endogenous pigment or enzyme activity before assigning residual brown staining to TNFRSF10B (general IHC practice).
How should I score TNFRSF10B staining across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because TNFRSF10B is membrane-annotated while HPA reports cytoplasmic tissue staining (UniProt O14763 subcellular annotation; HPA tissue IHC). For comparable sections, record % positive cells and staining intensity as an H-score, reporting membrane-associated and cytoplasmic results separately (general IHC practice). Normalise positive-cell counts to all evaluable cells of the same type, or report positive-cell density per mm² of viable tissue (general IHC practice). Keep section thickness, imaging and DAB development consistent, and exclude folds, edges and necrosis using the same prespecified rules for every specimen (general IHC practice).
What distinguishes credible TNFRSF10B staining from a misleading positive result?
A credible result follows a reproducible pattern in preserved cells and exceeds no-primary background under matched detection conditions (general IHC practice). Check whether signal is membrane-associated or cytoplasmic: UniProt annotates TNFRSF10B as membrane-localised, whereas HPA reports ubiquitous cytoplasmic tissue staining (UniProt O14763 subcellular annotation; HPA tissue IHC). Assess the stained cell type explicitly, since HPA lists high staining in several cell populations but reports only medium consistency with RNA data (HPA tissue IHC). Treat staining confined to section edges, necrotic regions or sites of endogenous enzyme activity as suspect, and confirm its pattern on adjacent sections before interpreting it biologically (general IHC practice).
Boster reagents
Best TNFRSF10B / Tumor necrosis factor receptor superfamily member 10B IHC Antibodies
Catalog images show TNFRSF10B staining in human, mouse and rat paraffin sections, plus IF in human, mouse and rat samples (catalog image captions).
Real IHC data
Anti-DR5/TNFRSF10B Antibody ®
Cat # A00410
Real IHC data
Anti-DR5 TNFRSF10B Antibody
Cat # A00410-2
Real IHC data
Anti-DR5/TNFRSF10B Antibody ®
Cat # A00410-5
Real IHC data
Anti-DR5/TNFRSF10B Antibody ®
Cat # PA1842
A00410 has IHC images from rat kidney, mouse intestine and human intestinal cancer; A00410-2 has mouse kidney IHC and human, mouse and rat IF images (catalog image captions). A00410-5 has IHC images from human endometrioid adenocarcinoma, lung cancer and tonsil; PA1842 shows human lung cancer IHC (catalog image captions).
Which to pick: For human tissue IHC, A00410-5 shows paraffin-section staining in three human tissues, with fixative unreported; A00410-2 shows paraffin-section mouse kidney paraffin sections (respective IHC image captions). For cross-species IHC, A00410 lists human, mouse and rat reactivity and shows paraffin sections from all three, with fixative unreported (catalog reactivity; A00410 IHC image captions). For IF/ICC, choose A00410-2: it lists IF for human, mouse and rat and shows 4% paraformaldehyde-fixed human HepG2 cells; clonality is unreported for every SKU (catalog applications and clone fields; A00410-2 IF image captions). The selected A00410 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A00410).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14763 (TR10B_HUMAN, Tumor necrosis factor receptor superfamily member 10B).
- Human Protein Atlas. TNFRSF10B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TNFRSF10B subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. TNFRSF10B antibody validation summary (1 antibodies).
- TNFRSF10B Implicated in Osteoarthritis Protection via the Alpha-Tocopherol-to-Sulfate Ratio: A Multiomics and Mendelian Randomization Study. International Journal of Molecular Sciences 2026 — PMC13607894.
- THE CRITICAL ROLE OF THE HISTONE MODIFICATION ENZYME SETDB2 IN THE PATHOGENESIS OF ACUTE RESPIRATORY DISTRESS SYNDROME. Shock (Augusta, Ga.) 2023 — PMC10417228.
- System-wide identification of novel de-ubiquitination targets for USP10 in gastric cancer metastasis through multi-omics screening. BMC cancer 2024 — PMC11209979.
- Targeting the miRNA-155/TNFSF10 network restrains inflammatory response in the retina in a mouse model of Alzheimer's disease. Cell death & disease 2021 — PMC8492692.
- PubMed PMID:9285725 — UniProt-cited evidence.
- PubMed PMID:9311998 — UniProt-cited evidence.
- PubMed PMID:9373179 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TNFRSF10B (Tumor necrosis factor receptor superfamily member 10B). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); membrane topology (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00410). Validated IHC antibody: A00410 at 0.5-1μg/ml (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-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A00410); 2. Fixation: Image fixative and duration unreported (datasheet A00410); 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 A00410); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00410); 8. Primary antibody: Rabbit anti-TNFRSF10B, 0.5-1μg/ml (datasheet A00410); 9. Primary incubation: Overnight at 4 °C (datasheet A00410). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining may differ from RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope side matters; coverage unknown (UniProt); Regulation: HCMV UL141 blocks surface expression (UniProt). Evidence: UniProt O14763, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A00410, PMC13607894, PMC10417228. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TNFRSF10B/immunohistochemistry-ihc-guide.