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TUSC3 / Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit TUSC3 · IHC design guide
Design Immunohistochemistry for TUSC3
For paraffin-section IHC, start with the catalog antibody at 1:100–1:300 (datasheet A05834). Assess cytoplasmic and membranous staining against the high staining reported in cervical or esophageal squamous cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TUSC3 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | High staining in squamous epithelia; cytoplasmic and membranous (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05834) | |
| Positive control | Cervix+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 conditions consistent across sections (standard IHC practice; not target-specific); target-specific effects are unknown. | |
| Caveat | The antibody may detect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Broad non-lymphoid expression (UniProt) | |
| Isoform / epitope | 2 isoforms; map epitopes to ER-lumenal or cytoplasmic regions (UniProt) |
Section 1
Recommended TUSC3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published TUSC3 staining protocols for paraffin tissue sections (PMC4752499; PMC5825997; PMC5581085; PMC6507427).
| Sample | Paraffin-embedded human Squamous cell carcinoma of lung tissue; fixative not specified (datasheet A05834) |
| Fixation | Image fixative and duration unreported (datasheet A05834); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A05834); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-TUSC3, 1:100-1:300 (datasheet A05834) |
| 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 | TUSC3-positive staining in squamous epithelial cells of cervix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 heat retrieval for the catalog antibody (datasheet A05834); consider citrate pH 6.0 when adapting the published liver protocol (PMC5825997).
Section 2
What Is the Expected TUSC3 Staining Pattern?
TUSC3 is an endoplasmic reticulum membrane protein with 4 transmembrane segments (UniProt Q13454 topology). In paraffin sections, expect cytoplasmic and membranous staining, particularly in squamous epithelial cells and selected glandular cells (HPA: tissue IHC). HPA rates the tissue staining “Approved,” with medium consistency against RNA data, and cautions that its antibodies target protein from more than one gene (HPA: reliability description).
| Cytoplasmic and membranous staining is strong in esophageal squamous epithelial cells or small intestinal glandular cells (HPA: High in both cell populations). | This fits the reported tissue pattern and an endoplasmic reticulum membrane location (HPA: tissue IHC; UniProt Q13454 subcellular location). Record the stained cell population as well as intensity; a positive area alone does not establish which cells express TUSC3 (standard IHC practice). |
| Staining is confined to nuclei, with no convincing cytoplasmic or membranous signal. | A nucleus-only result conflicts with the reported compartment pattern and membrane topology (HPA: tissue IHC; UniProt Q13454 topology). Treat it as suspect; compare the same run with a known-positive tissue and review counterstain, detection and background before assigning TUSC3 localisation (standard IHC practice). |
| Strong staining appears in adipocytes or bone marrow hematopoietic cells (HPA: Not detected in these cell populations). | That distribution conflicts with the reported tissue observations (HPA: tissue IHC). Cross-reactivity or endogenous detection activity is possible; HPA also cautions that its antibodies target protein from more than one gene (HPA: reliability description). Check control slides before calling these cells positive (standard IHC practice). |
| Diffuse colour covers tissue and empty spaces, obscuring cell boundaries. | The distribution cannot be scored reliably as cytoplasmic or membranous staining (HPA: tissue IHC pattern; standard IHC practice). Uneven blocking, residual detection activity or excessive chromogen development are general IHC possibilities; this appearance does not identify a TUSC3-specific cause (standard IHC practice). |
| No staining is seen in esophageal squamous epithelium processed alongside the study sections (HPA: High in squamous epithelial cells). | The expected positive control has failed, so negative study sections are inconclusive (HPA: tissue IHC; standard IHC practice). Review tissue preservation, retrieval, primary antibody and detection steps as general workflow checks; no target-specific fixation sensitivity is established by the supplied record. |
💡Expected TUSC3 appearanceCall a section positive when its named cell population shows a clear cytoplasmic and membranous pattern, potentially High in esophageal squamous epithelium or small intestinal glands; nucleus-only staining or strong signal in reported negative cells is suspect (HPA: tissue IHC; UniProt Q13454 subcellular location).
| Cell population and tissue | HPA reports High staining in cervical and esophageal squamous epithelial cells, skin keratinocytes and small intestinal glandular cells, but Not detected in adipocytes (HPA: tissue IHC). Compare like cell populations when choosing controls; whole-organ labels can hide these differences (standard IHC practice). |
| Membrane topology and epitope position | TUSC3 has 4 transmembrane segments and lumenal and cytoplasmic regions (UniProt Q13454 topology). The supplied record does not map either antibody's epitope, so topology alone cannot determine an antibody-specific retrieval or permeabilisation setting. |
| Processing and isoforms | UniProt lists a signal sequence at residues 1–41, a chain at 42–348 and 2 isoforms (UniProt Q13454 processing and isoforms). Without an epitope map or isoform-specific validation, staining cannot identify which isoform was detected or whether an antibody recognises the processed chain. |
| Strength of IHC evidence | Both listed antibodies, HPA049851 and HPA049974, are IHC “Approved”; neither is reported here as IHC “Enhanced” (HPA: antibody validation). The tissue profile has medium consistency with RNA data and a multiple-gene target caution, so its levels are useful reference observations rather than proof of antibody specificity (HPA: reliability description). |
| IF/ICC: should the IHC pattern be carried over? | HPA's ICC-IF summary says “Membrane,” but gives no main location or cell-line images; ICC status is unlisted for both supplied antibodies (HPA: subcellular record and antibody validation). Treat that as limited IF evidence. This section provides no IF/ICC protocol or prediction of a validated cell-line pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive esophageal squamous epithelium is blank (HPA: High in squamous epithelial cells). | A run or reagent problem is possible; the supplied sources do not establish TUSC3-specific fixation sensitivity (standard IHC practice). | Repeat with a positive control and verify retrieval, primary antibody, detection reagent and chromogen steps against the chosen IHC protocol (standard IHC practice). |
| The slide shows only nuclear colour. | This conflicts with the cytoplasmic and membranous pattern and endoplasmic reticulum membrane location (HPA: tissue IHC; UniProt Q13454). | Check a positive-control section and a no-primary control; reassess counterstain and background before scoring the nuclei (standard IHC practice). |
| Adipocytes stain strongly (HPA: Not detected in adipocytes). | Cross-reactivity or endogenous detection activity is possible; HPA warns of protein targeting from more than one gene (HPA: reliability description; standard IHC practice). | Compare a no-primary control and the expected positive tissue in the same run; record the unexpected cell-specific pattern (standard IHC practice). |
| Colour is widespread and cell borders cannot be resolved. | General IHC background or excessive chromogen development can obscure the reported compartment pattern (standard IHC practice; HPA: tissue IHC). | Review blocking, washes and chromogen development; score only after cell boundaries and compartments are distinguishable (standard IHC practice). |
| A low-staining glandular population is called negative. | HPA reports Low staining in thyroid glandular cells and pancreatic exocrine glandular cells; faint signal may be missed (HPA: tissue IHC). | Compare the named cells with a high-staining control under the same scoring conditions, and report weak and absent signal separately (HPA: tissue IHC; standard IHC practice). |
| An ICC-IF result is used to settle an ambiguous paraffin-section result. | HPA supplies a “Membrane” ICC-IF summary without cell-line images or ICC validation for the listed antibodies (HPA: subcellular record and antibody validation). | Interpret the paraffin section against its IHC controls and named tissue pattern; describe any IF observation separately (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing TUSC3 staining
A05834 · IHCImmunohistochemical analysis of paraffin-embedded human Squamous cell carcinoma of lung. 1, Antibody was diluted at 1:200 (4° overnight). 2,
A05834 · IFImmunofluorescence analysis of HeLa cells using TUSC3 Antibody. The picture on the right is blocked with the synthesized peptide.
A05834-1 · IHCImmunohistochemistry (IHC) analyzes of TUSC3 (H153) pAb in paraffin-embedded human tonsil carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104723-TUSC3/tissue/cerebral+cortex#imid_14848376
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104723-TUSC3/tissue/colon#imid_14848302
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104723-TUSC3/tissue/liver#imid_14848303
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000104723-TUSC3/tissue/kidney#imid_14848304
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TUSC3 IHC & IF
🧪Run cervix first; its squamous epithelial cells should stain (HPA: High in cervix squamous epithelial cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes), and treat unstained areas beside the positive squamous cells as internal background controls without assuming those cells lack TUSC3 (HPA: no cell-level negative specified for cervix).
Positive control tissue: Cervix (Squamous epithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for TUSC3; derive a cell-line control from the positive tissue's cell type (Squamous epithelial cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and immunoglobulin class; and a TUSC3 knockout biological control where available (standard IHC practice). Block endogenous peroxidase in the cervix section and check for staining at tissue edges (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A05834 paraffin-section caption does not state a fixative (selected SKU tissue-IHC caption). The reported IHC condition uses Tris-EDTA retrieval at pH 9.0, so retrieval should be checked before scoring (selected SKU tissue-IHC caption); the supplied evidence does not establish whether frozen sections or IF are easier (HPA subcellular: no ICC-IF cell-line images). In cervix squamous epithelium, assess tissue-edge staining separately from staining within intact cells (standard chromogenic IHC practice).
HPA tissue IHC evidence for TUSC3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TUSC3 IHC Tips
Use the catalog antibody’s paraffin-section image as a starting point, then assess staining by cell type, compartment, and controls (caption A05834; HPA tissue IHC; UniProt Q13454 localisation).
Which retrieval condition should I start with for TUSC3 staining in paraffin sections?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 for paraffin sections (datasheet A05834). The selected tissue image used primary antibody at 1:200 at 4°C overnight after this retrieval (caption A05834). Include a stained control and a no-primary control when comparing retrieval conditions (standard IHC practice). If staining is weak, test a milder retrieval condition on matched sections while keeping antibody and detection settings constant (standard IHC practice). Judge improvement by cellular staining rather than a stronger edge or background signal; TUSC3 is an ER membrane protein (UniProt Q13454 localisation; standard IHC practice).
Could fixation explain weak or uneven TUSC3 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A05834). Record the fixative, fixation duration, processing history, and section age before attributing weak staining to antigen loss (standard IHC practice). Compare suspect sections with a similarly processed positive control using Tris-EDTA at pH 9.0 and the same detection run (datasheet A05834; standard IHC practice). If tissue edges stain more strongly than the center, check processing consistency and section adhesion before increasing antibody concentration (standard IHC practice). HPA staining patterns and TUSC3 topology do not establish how this target responds to fixation (HPA tissue IHC; UniProt Q13454 topology).
Where should convincing TUSC3 staining appear within a positive cell?
Assess a cytoplasmic or membranous pattern first, as reported across tissues by HPA (HPA tissue IHC). TUSC3 resides in the ER membrane and has 4 transmembrane segments, so a predominantly nuclear chromogenic signal warrants scrutiny (UniProt Q13454 localisation and topology). Compare staining with intact cell outlines and adjacent unstained areas on the same section before assigning it to a compartment (standard IHC practice). The catalog image documents staining in paraffin-embedded human lung squamous cell carcinoma, without establishing a universal cell-type pattern (caption A05834). Use a no-primary control and an independent positive tissue control when compartment assignment is ambiguous (standard IHC practice).
How can epitope placement affect interpretation of TUSC3 staining?
The record lists 2 TUSC3 isoforms, but the supplied antibody evidence does not identify which isoform or epitope is detected (UniProt Q13454 isoforms; caption A05834). TUSC3 has a lumenal region at residues 42–196 and several membrane-spanning segments, making epitope accessibility a practical concern (UniProt Q13454 topology). Avoid calling a negative section isoform-specific without an antibody epitope map and appropriate validation (standard IHC practice). Apply the catalog retrieval condition, Tris-EDTA at pH 9.0, consistently across comparisons (datasheet A05834). If patterns disagree between antibodies, compare their stated epitopes and controls before interpreting biological differences (standard IHC practice).
How should I adapt the TUSC3 question for multiplex IF?
For a separate IF experiment, pair TUSC3 with a marker for the expected cell population, such as a squamous epithelial marker when assessing the HPA-positive squamous cells (HPA tissue IHC; standard IF practice). Select fluorophores and imaging channels after checking tissue autofluorescence with an unstained control (standard IF practice). Choose permeabilisation after locating the antibody epitope: TUSC3 has lumenal regions and short cytoplasmic regions separated by 4 transmembrane segments (UniProt Q13454 topology; standard IF practice). The supplied caption validates a paraffin-section chromogenic image, with retrieval at pH 9.0; it supplies no IF fixation or permeabilisation result (caption A05834). Assess colocalisation against single-stain and no-primary controls (standard IF practice).
What should I change when TUSC3 chromogen spreads beyond cells?
First compare the stained section with a no-primary control to identify detection-related background (standard IHC practice). Check peroxidase blocking before chromogen development, then inspect whether diffuse signal tracks tissue edges, folds, or damaged regions (standard IHC practice). The selected image used primary antibody at 1:200 at 4°C overnight and secondary antibody at 1:200 for 45 minutes at room temperature (caption A05834). If controls implicate nonspecific binding, optimize blocking and antibody concentration one variable at a time (standard IHC practice). Treat uniform nuclear deposits cautiously because TUSC3 is an ER membrane protein (UniProt Q13454 localisation; standard IHC practice).
How should I score TUSC3 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; HPA reports cytoplasmic and membranous staining across most tissues (HPA tissue IHC). For comparable regions, record percent positive cells and intensity, then calculate an H-score from intensity categories 0–3 if that scale fits the study (standard IHC practice). Normalize counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue (standard IHC practice). Keep retrieval, detection, counterstain, exposure during image capture, and threshold rules consistent across sections (standard IHC practice). Exclude folds, necrosis, and section edges using prespecified criteria, and retain controls with each staining run (standard IHC practice).
When is a positive TUSC3 stain likely to be an artefact?
A convincing result should have interpretable cellular boundaries and a cytoplasmic or membranous pattern, consistent with HPA tissue staining and ER membrane localisation (HPA tissue IHC; UniProt Q13454 localisation). HPA reports high staining in cervical squamous epithelial cells but no detected staining in lung alveolar cells, so identify the stained cell type before comparing tissues (HPA tissue IHC). Discount signal confined to edges, folds, or necrotic areas when corresponding intact tissue lacks staining (standard IHC practice). Investigate deposits that persist without primary antibody or after inadequate peroxidase blocking as detection artefacts (standard IHC practice). HPA rates its tissue staining Approved but cautions that its antibody targets protein from more than one gene, limiting target-specific conclusions (HPA tissue IHC).
Boster reagents
Best TUSC3 / Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit TUSC3 IHC Antibodies
Two anti-TUSC3 antibodies have human paraffin-section IHC images; A05834 also has a HeLa-cell IF image, and both list Human, Mouse and Rat reactivity (IHC/IF image captions; catalog reactivity).
Real IHC data
Anti-N33 TUSC3 Antibody
Cat # A05834
Real IHC data
Anti-TUSC3 (H153) Antibody
Cat # A05834-1
A05834 will render with an IHC image of paraffin-embedded human lung squamous cell carcinoma and has an IF image in HeLa cells (A05834 image captions). A05834-1 will render with an IHC image of paraffin-embedded human tonsil carcinoma; its listed applications do not include IF (A05834-1 IHC image caption; catalog applications).
Which to pick: For tissue IHC, A05834 has a documented paraffin-section example using Tris-EDTA retrieval at pH 9.0; A05834-1 has a paraffin-section example, and neither caption reports the fixative (IHC image captions). For IF/ICC, choose A05834 because IF is listed and illustrated in HeLa cells (catalog applications; A05834 IF image caption). For cross-species work, both are rabbit polyclonals listing Human, Mouse and Rat reactivity, though their shown IHC examples are human tissue (catalog host, dilution_raw and reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13454 (TUSC3_HUMAN, Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit TUSC3).
- Human Protein Atlas. TUSC3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TUSC3 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. TUSC3 antibody validation summary (2 antibodies).
- Decreased TUSC3 Promotes Pancreatic Cancer Proliferation, Invasion and Metastasis. PloS one 2016 — PMC4752499.
- Low levels of tumor suppressor candidate 3 predict poor prognosis of patients with hepatocellular carcinoma. OncoTargets and therapy 2018 — PMC5825997.
- TUSC3 induces autophagy in human non-small cell lung cancer cells through Wnt/β-catenin signaling. Oncotarget 2017 — PMC5581085.
- TUSC3 as a potential biomarker for prognosis in clear cell renal cell carcinoma. Oncology letters 2019 — PMC6507427.
- PubMed PMID:8661104 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TUSC3 (Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit TUSC3). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05834). Validated IHC antibody: A05834 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Squamous epithelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Squamous cell carcinoma of lung tissue; fixative not specified (datasheet A05834); 2. Fixation: Image fixative and duration unreported (datasheet A05834); 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: Tris-EDTA pH 9.0 (datasheet A05834); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-TUSC3, 1:100-1:300 (datasheet A05834); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); target-specific effects are unknown.; Caveat: The antibody may detect protein from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; map epitopes to ER-lumenal or cytoplasmic regions (UniProt); Regulation: Broad non-lymphoid expression (UniProt). Evidence: UniProt Q13454, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A05834, PMC4752499, PMC5825997. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TUSC3/immunohistochemistry-ihc-guide.