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TNS3 / Tensin-3 · IHC design guide
Design Immunohistochemistry for TNS3
Plan TNS3 IHC in paraffin sections using glandular cells with high reported staining as positive references (HPA tissue IHC). Compare membranous and cytoplasmic staining while accounting for the reported low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TNS3 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 | Membranous and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Membranous/cytoplasmic staining in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07345-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low consistency with RNA data (HPA tissue IHC) | |
| Regulation | EGF down-regulates TNS3 (UniProt) | |
| Isoform / epitope | 4 isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended TNS3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with 4 published TNS3 IHC protocols (PMC9392296; PMC8146480; PMC8221360; PMC13048582).
| Sample | Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A07345-1) |
| Fixation | Image fixative and duration unreported (datasheet A07345-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A07345-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07345-1) |
| Primary antibody | Rabbit anti-TNS3, 2-5 μg/ml (datasheet A07345-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07345-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07345-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TNS3-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Membranous and cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A07345-1). Use citrate pH 6.0 when reproducing the published citrate protocols (PMC8146480; PMC13048582).
Section 2
What Is the Expected TNS3 Staining Pattern?
TNS3 localizes to focal and fibrillar adhesions and podosomes (UniProt Q68CZ2); it has no transmembrane segment (UniProt Q68CZ2 topology). In paraffin sections, expect membranous and cytoplasmic staining in selected glandular and other positive cell populations (HPA tissue IHC). Treat tissue predictions cautiously: HPA rates its IHC profile Approved but reports low consistency between antibody staining and RNA expression, pending external verification (HPA tissue IHC).
| Membranous and cytoplasmic signal in colon, appendix, duodenum or gallbladder glandular cells. | This matches the reported tissue pattern; HPA scores these cell populations High (HPA tissue IHC). Assess the named cells within the section, rather than treating a uniformly stained field as confirmation. HPA's Approved profile still has low staining–RNA consistency, so this pattern supports interpretation without independently proving antibody specificity (HPA tissue IHC). |
| Predominantly nuclear staining, with little signal at cell edges or in cytoplasm. | A nuclear-only pattern conflicts with the reported membranous and cytoplasmic tissue profile (HPA tissue IHC) and adhesion-associated localization (UniProt Q68CZ2). Suspect nonspecific staining or a detection artefact; compare positive and negative control sections before assigning biological meaning. Adhesion-scale puncta may be difficult to resolve by chromogenic tissue IHC (general IHC practice). |
| Strong signal in adipocytes, cardiomyocytes or liver cholangiocytes. | Those specific populations are listed as Not detected (HPA tissue IHC). Check the cell identification and staining controls; cross-reactivity or endogenous detection activity could explain unexpected signal (general IHC practice). HPA's low staining–RNA consistency means an unexpected result warrants verification, rather than automatic rejection as impossible (HPA tissue IHC). |
| Diffuse stain spans cells and extracellular space without a discernible cellular pattern. | This does not resemble the reported membranous and cytoplasmic expression (HPA tissue IHC). In chromogenic IHC, incomplete blocking, excess primary antibody or residual detection background can produce such appearances (general IHC practice). Review a no-primary control and the section's morphology before scoring diffuse colour as TNS3. |
| No signal in glandular cells of a well-preserved colon or appendix section. | HPA reports High staining in those populations (HPA tissue IHC), so first investigate a technical false negative using the assay's controls (general IHC practice). Recheck that the relevant glandular cells are present, then review the antibody dilution, retrieval conditions and detection run. HPA's Approved rating is pending external verification (HPA tissue IHC). |
💡Expected TNS3 appearanceCall a result positive when identifiable glandular cells show High membranous and cytoplasmic staining in colon or appendix (HPA tissue IHC); isolated nuclear colour or uniform extracellular colour is inconsistent with that reported pattern and should prompt control review (HPA tissue IHC; general IHC practice).
| Subcellular location | TNS3 is associated with focal and fibrillar adhesions, predominantly the latter, and podosomes (UniProt Q68CZ2). HPA describes tissue staining as membranous and cytoplasmic (HPA tissue IHC). A chromogenic section need not resolve individual adhesion sites to agree with those observations (general IHC practice). |
| Cell population and tissue | HPA reports High staining in kidney glomerular cells, pancreatic exocrine glandular cells and fallopian tube ciliated-cell rootlets, alongside several glandular populations (HPA tissue IHC). Select and score the stated cell population; a tissue name alone does not define where signal should appear (general IHC practice). |
| Antibody evidence | Both listed antibodies, HPA055338 and HPA056015, have Approved IHC status (HPA antibodies). The tissue profile nevertheless has low consistency with RNA expression and awaits external verification (HPA tissue IHC). Interpret staining with morphology and controls; Approved status alone does not make every unexpected pattern specific. |
| Isoforms and epitope | UniProt lists 4 TNS3 isoforms and domains spanning different parts of the protein (UniProt Q68CZ2). Without an epitope location in these payloads, the isoforms recognized by a particular antibody cannot be predicted. Do not attribute a staining difference to isoform selection from these data alone. |
| IF/ICC interpretation | If an IF/ICC image is being compared with the IHC slide, HPA places TNS3 mainly at focal adhesion sites and lists Supported ICC status for both antibodies (HPA subcellular; HPA antibodies). The IF localization offers a compartment check, but its spatial detail should not be required of chromogenic IHC (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control glandular cells are blank. | The assay may have lost detectable signal; colon and appendix glandular cells are reported High (HPA tissue IHC). | Confirm the expected cells are present, then review the IHC-validated antibody's specified dilution, retrieval and detection controls (general IHC practice). These sources do not establish a TNS3-specific fixation sensitivity. |
| Signal appears chiefly in nuclei. | That compartment conflicts with HPA's membranous and cytoplasmic tissue profile (HPA tissue IHC). | Check the no-primary control, section morphology and detection background; repeat with an independently validated antibody when available (general IHC practice). Avoid calling nuclear-only colour a confirmed TNS3 pattern. |
| Negative-reference cells stain strongly. | Adipocytes and cardiomyocytes are reported Not detected, so background, endogenous activity or cross-reactivity is plausible (HPA tissue IHC; general IHC practice). | Verify cell identity and compare no-primary and appropriate detection controls before changing interpretation (general IHC practice). HPA's staining–RNA discordance calls for verification of unexpected biology (HPA tissue IHC). |
| Brown colour is diffuse across the section. | A field-wide haze lacks the reported cell-associated membranous and cytoplasmic pattern (HPA tissue IHC); excess reagent or inadequate background control can contribute (general IHC practice). | Inspect the no-primary control, blocking and washes; adjust assay conditions within the antibody's documented IHC procedure (general IHC practice). Score only interpretable cellular staining. |
| Signal varies between neighboring cell types. | HPA assigns levels to specific populations, such as High glomerular cells and Not detected liver cholangiocytes, rather than to every cell in an organ (HPA tissue IHC). | Annotate morphology and score each relevant population separately (general IHC practice). Check the HPA cell-level expectation before treating heterogeneity as assay failure (HPA tissue IHC). |
| Does IF/ICC have to show the same appearance as chromogenic IHC? | HPA reports focal adhesion localization by ICC-IF, while its tissue IHC summary is membranous and cytoplasmic (HPA subcellular; HPA tissue IHC). | Use the IF observation as a localization cross-check, then judge the paraffin IHC slide against its own cell-level tissue pattern and controls (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing TNS3 staining
A07345-1 · IHCIHC analysis of TNS3 using anti-TNS3 antibody (A07345-1). TNS3 was detected in a paraffin-embedded section of human colon adenocarcinoma tis
A07345-1 · IHCIHC analysis of TNS3 using anti-TNS3 antibody (A07345-1). TNS3 was detected in a paraffin-embedded section of human endometrioid adenocarcin
A07345-1 · IFIF analysis of TNS3 using anti-TNS3 antibody (A07345-1). TNS3 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen re
A30511 · IHCImmunohistochemistry analysis of paraffin-embedded human heart tissue, using TENS3 Antibody. The picture on the right is blocked with the sy
A30511 · IFImmunofluorescence analysis of COS7 cells, using TENS3 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136205-TNS3/tissue/cerebral+cortex#imid_17390539
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136205-TNS3/tissue/colon#imid_17406251
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136205-TNS3/tissue/liver#imid_17449446
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000136205-TNS3/tissue/pancreas#imid_17449397
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055338; green = TNS3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA055338; green = TNS3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TNS3 IHC & IF
🧪Run colon first and expect high staining in glandular cells (HPA: High in colon glandular cells). Run adipose tissue as a negative comparator and expect adipocytes to remain unstained (HPA: Not detected in adipocytes); on the colon slide, assess neighboring cells without glandular staining for counterstain-only background rather than assuming a particular cell type is negative (HPA: colon glandular-cell row).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TNS3 in A-549, U-251MG, U2OS, A-431, with annotated localisation: Focal adhesion sites (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched nonimmune rabbit IgG control of the same antibody format (selected-SKU caption: rabbit anti-TNS3; standard IHC practice). Use a TNS3 knockout specimen as a biological negative where available, and quench endogenous peroxidase in colon sections before HRP/DAB detection (standard IHC practice; selected-SKU caption: HRP/DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative unreported). The caption uses heat retrieval in EDTA, pH 8.0, but does not establish that retrieval is required or that frozen sections or IF are easier (selected-SKU caption: EDTA retrieval; HPA: ICC-IF images available). In colon, assess glandular staining against possible mucin-associated and endogenous peroxidase background (standard IHC practice).
HPA tissue IHC evidence for TNS3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TNS3 IHC Tips
Troubleshoot TNS3 staining in paraffin-section chromogenic IHC by checking retrieval, controls, compartment-specific signal and scoring before interpreting tissue differences.
Which retrieval conditions should I use when TNS3 staining is weak?
Start with heat-mediated retrieval in EDTA buffer at pH 8.0 for this paraffin-section IHC assay (datasheet A07345-1). The selected image used that retrieval before 2 μg/ml primary antibody overnight at 4°C, so reproduce those linked conditions before changing the buffer (datasheet A07345-1). If staining remains weak, compare shorter and longer heating steps on matched sections, then consider another buffer only as a documented fallback (standard IHC practice). Keep section thickness, cooling, antibody concentration and DAB development matched; assess signal in colon glandular cells alongside a no-primary control (HPA: High in colon glandular cells; standard IHC practice).
How should I assess whether fixation caused a weak TNS3 result?
The selected image shows a paraffin-embedded colon adenocarcinoma section, but its fixative is unreported; TNS3-specific fixation sensitivity is therefore unknown (datasheet A07345-1). Record the fixative and fixation duration for each specimen, and compare sections processed together before attributing a weak result to fixation (standard IHC practice). Hold retrieval at EDTA pH 8.0 and the primary antibody at 2 μg/ml overnight at 4°C during that comparison (datasheet A07345-1). Include an expected positive compartment and a no-primary control on the same run to separate lost signal from detection background (HPA: High in colon glandular cells; standard IHC practice).
What staining pattern is plausible for TNS3 in tissue sections?
Assess TNS3 primarily in cytoplasmic and membrane-associated compartments, the pattern reported across several tissues by tissue IHC (HPA: membranous and cytoplasmic expression). TNS3 localizes to cell junctions and podosomes and favors fibrillar over focal adhesions, so interface-associated signal is biologically plausible (UniProt Q68CZ2: subcellular location). Because it has no transmembrane segment, a sharp membrane outline alone should not establish specificity without supporting controls (UniProt Q68CZ2: topology; standard IHC practice). In colon, score glandular cells separately from surrounding tissue and compare the pattern with a no-primary section (HPA: High in colon glandular cells; standard IHC practice).
Could isoforms or epitope accessibility explain discordant TNS3 staining?
TNS3 has 4 annotated isoforms, so establish which sequence the antibody recognizes before interpreting a negative section as absence of all TNS3 (UniProt Q68CZ2: isoforms; standard IHC practice). Its tensin-type phosphatase and C2 domains lie near the N terminus, while SH2 and PTB domains lie near the C terminus (UniProt Q68CZ2: domains). Map the immunogen to those regions and check whether the relevant sequence is shared by the isoforms under study (UniProt Q68CZ2: isoforms and domains; standard IHC practice). If epitope mapping is unavailable, report staining as antibody-defined TNS3 immunoreactivity and compare independently validated reagents where feasible (standard IHC practice).
How can I investigate TNS3 localisation with IF alongside this IHC assay?
Treat IF/ICC as a separate assay: the selected antibody caption establishes paraffin-section chromogenic IHC conditions, not IF conditions (datasheet A07345-1). Multiplex TNS3 with a marker for the expected cell type, such as an epithelial marker when examining glandular cells, and include single-label controls (HPA: High in colon glandular cells; standard IF practice). Choose spectrally separated fluorophores and favor a far-red channel when specimen autofluorescence obscures shorter wavelengths (standard IF practice). TNS3 lacks a transmembrane segment and associates with adhesions, so use gentle permeabilisation for access to intracellular epitopes, then verify the pattern against no-primary controls (UniProt Q68CZ2: topology and subcellular location; standard IF practice).
How do I reduce diffuse brown staining without losing TNS3 signal?
Begin with matched no-primary and secondary-only sections to locate background introduced by detection reagents (standard IHC practice). The selected IHC image used 10% goat serum blocking and a peroxidase-conjugated secondary for 30 minutes at 37°C; reproduce those documented conditions while troubleshooting (datasheet A07345-1). Quench endogenous peroxidase before HRP/DAB detection, and adjust wash stringency or primary concentration one variable at a time (standard IHC practice). Compare any improvement in background with retained signal in expected colon glandular cells; an evenly darker section is insufficient evidence of stronger TNS3 staining (HPA: High in colon glandular cells; standard IHC practice).
How should I quantify TNS3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic or membrane-associated compartment before scoring, because tissue IHC reports both staining patterns for TNS3 (HPA: membranous and cytoplasmic expression; standard IHC practice). Record the percentage of viable target cells at each intensity and calculate an H-score from 0–300, or report percentage positive when intensity cannot be graded consistently (standard IHC practice). For spatial comparisons, measure positive-cell density per mm² of viable, annotated tissue and exclude necrotic or folded regions (standard IHC practice). Normalize comparisons to the same cell type, tissue area, detection settings and scoring threshold across sections (standard IHC practice).
Which findings distinguish credible TNS3 staining from artefact?
Credible signal should occur in an expected cell population and a plausible cytoplasmic or membrane-associated compartment; colon glandular cells provide one documented comparison (HPA: High in colon glandular cells; HPA: membranous and cytoplasmic expression). Adhesion-associated staining fits TNS3 biology, whereas isolated nuclear staining requires separate validation before interpretation (UniProt Q68CZ2: subcellular location; standard IHC practice). Discount staining concentrated at section edges, folds or necrotic regions, and check whether a no-primary section reveals residual endogenous-enzyme signal (standard IHC practice). Interpret intensity cautiously because the tissue IHC resource reports low consistency between antibody staining and RNA expression and lists its reliability as Approved pending external verification (HPA: reliability description).
Boster reagents
Best TNS3 / Tensin-3 IHC Antibodies
Both antibodies have human paraffin-section IHC images and cell IF images (catalog image captions). Listed reactivity is human for A07345-1 and human, mouse, and rat for A30511 (catalog reactivity).
Real IHC data
Anti-TNS3 Antibody ®
Cat # A07345-1
Real IHC data
Anti-TENS3 Antibody
Cat # A30511
A07345-1 shows IHC in paraffin-embedded human colon adenocarcinoma and IF in U2OS cells (A07345-1 image captions). A30511 shows IHC in paraffin-embedded human heart and IF in COS7 cells, each with a peptide-blocked comparison (A30511 image captions).
Which to pick: For tissue IHC, choose A07345-1 when its documented EDTA pH 8.0 retrieval and 2 μg/ml overnight incubation fit your workflow; A30511 has a peptide-blocked human heart IHC image but fewer processing details (respective IHC captions). For IF/ICC, both have cell images; choose A30511 if listed mouse or rat reactivity matters, while its illustrated IHC sample is human (catalog reactivity; respective image captions). The fixative is unreported for both paraffin-section IHC images (respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q68CZ2 (TENS3_HUMAN, Tensin-3).
- Human Protein Atlas. TNS3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TNS3 subcellular location (ICC-IF): Mainly localized to the focal adhesion sites..
- Human Protein Atlas. TNS3 antibody validation summary (2 antibodies).
- Abnormal TNS3 gene methylation in patients with congenital scoliosis. BMC musculoskeletal disorders 2022 — PMC9392296.
- Immunohistochemical Analysis of the Expression of Adhesion Proteins: TNS1, TNS2 and TNS3 in Correlation with Clinicopathological Parameters in Gastric Cancer. Biomolecules 2021 — PMC8146480.
- Pharmacological targeting of TNS3 with histone deacetylase inhibitor as a therapeutic strategy in esophageal squamous cell carcinoma. Aging 2021 — PMC8221360.
- TNS1 and TNS4 play a potential role in development of pancreatic ductal adenocarcinoma but not TNS2 and TNS3. Cell adhesion & migration 2026 — PMC13048582.
- PubMed PMID:11559528 — UniProt-cited evidence.
- PubMed PMID:15140944 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TNS3 (Tensin-3). Expected localisation Membranous and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07345-1). Validated IHC antibody: A07345-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon adenocarcinoma tissue; fixative not specified (datasheet A07345-1); 2. Fixation: Image fixative and duration unreported (datasheet A07345-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A07345-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07345-1); 8. Primary antibody: Rabbit anti-TNS3, 2-5 μg/ml (datasheet A07345-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07345-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA data (HPA tissue IHC); Isoform / epitope: 4 isoforms; check epitope coverage across variants (UniProt); Regulation: EGF down-regulates TNS3 (UniProt). Evidence: UniProt Q68CZ2, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the focal adhesion sites., Datasheet A07345-1, PMC9392296, PMC8146480. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TNS3/immunohistochemistry-ihc-guide.