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SEC63 / Translocation protein SEC63 homolog · IHC design guide
Design Immunohistochemistry for SEC63
Plan SEC63 paraffin IHC using the high staining in pancreatic exocrine glandular cells as a positive reference (HPA tissue IHC). This guide covers fixation consistency, chromogenic detection and scoring of the observed cytoplasmic pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SEC63 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 | General cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Exocrine glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04852-1) | |
| Positive control | Pancreas+4 more · see all | |
| Negative control | Oral mucosa+2 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). Selected-image fixative and duration unreported (datasheet A04852-1); verify before use. | |
| Caveat | Hepatocytes stain low despite reported high liver expression (HPA tissue IHC; UniProt) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 0 isoforms; epitopes may face the ER lumen or cytoplasm (UniProt) |
Section 1
Recommended SEC63 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A04852-1). Two published SEC63 IHC protocols provide additional tissue-specific examples (PMC3508994; PMC2655295).
| Sample | Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A04852-1) |
| Fixation | Image fixative and duration unreported (datasheet A04852-1); 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 A04852-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04852-1) |
| Primary antibody | Rabbit anti-SEC63, 2-5 μg/ml (datasheet A04852-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04852-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04852-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SEC63-positive staining in exocrine glandular cells of pancreas (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A04852-1); the published liver protocol used citrate pH 6 (PMC3508994).
Section 2
What Is the Expected SEC63 Staining Pattern?
SEC63 is an endoplasmic reticulum membrane protein with three transmembrane segments (UniProt Q9UGP8 topology). In paraffin sections, expect cytoplasmic staining in pancreatic exocrine glandular cells, salivary glandular cells and testicular Leydig cells, each scored High by HPA (HPA tissue IHC). HPA describes the overall tissue profile as general cytoplasmic expression and rates its agreement with RNA expression as Supported (HPA tissue IHC).
| Cytoplasmic staining in pancreatic exocrine or salivary glandular cells. | This fits the reported High staining in those cells (HPA tissue IHC) and the expected ER location (UniProt Q9UGP8; HPA subcellular). Assess intensity in the named cells rather than treating every cell in the section as an equivalent positive control (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic signal. | A nuclear dominant pattern conflicts with the reported ER location and general cytoplasmic IHC profile (UniProt Q9UGP8; HPA tissue IHC). Treat it as suspect; inspect controls and detection background before assigning it to SEC63 (general IHC practice). |
| Strong staining in oral mucosal squamous cells or parathyroid glandular cells. | HPA reports SEC63 as Not detected in these cell types (HPA tissue IHC). Check cell identity and controls; an unexpected signal may reflect cross-reactivity or endogenous detection activity (general IHC practice). HPA's result does not establish absolute absence in every specimen. |
| Diffuse color across cells, stroma and empty areas. | Color without a cell associated cytoplasmic pattern is poor evidence of SEC63 localization (UniProt Q9UGP8; HPA tissue IHC). Review blocking, washes, chromogen exposure and a control omitting primary antibody (general IHC practice). |
| No staining in pancreatic exocrine glandular cells. | That conflicts with HPA's High tissue IHC result for those cells (HPA tissue IHC). First check that the cells are present and the run's positive control worked; a negative slide alone cannot establish SEC63 absence (general IHC practice). |
💡Expected SEC63 appearanceCall a section positive when pancreatic exocrine or salivary glandular cells show clear cytoplasmic chromogen staining consistent with an ER protein (HPA tissue IHC; UniProt Q9UGP8); HPA scores these cells High, whereas nuclear dominant or cell independent color is suspect (HPA tissue IHC; general IHC practice).
| Choice of positive and comparison tissue | Pancreatic exocrine glandular cells and salivary glandular cells are reported High; oral mucosal squamous cells are Not detected (HPA tissue IHC). Compare the specified cell populations, because a tissue level label does not score every constituent cell (HPA tissue IHC). |
| Antibody evidence | HPA053295 and HPA060535 each have Supported IHC status (HPA antibodies). This supports using their reported patterns as references, but it does not supply a dilution or show that a different catalog antibody has the same specificity (HPA antibodies). |
| Epitope location and accessibility | SEC63 has three membrane spans, a lumenal segment at residues 91–188 and a cytoplasmic region at 210–760 (UniProt Q9UGP8 topology). Interpret a known antibody epitope against that map; the supplied evidence gives no epitope for the catalog antibody or SEC63-specific retrieval requirement. |
| Processing and variants | UniProt lists one chain spanning residues 1–760, no signal peptide, no propeptide and no isoforms in this record (UniProt Q9UGP8). These entries offer no basis to predict a shed SEC63 signal or an isoform-specific tissue pattern. |
| Retrieval and chromogenic detection | For paraffin IHC, use the IHC-validated antibody's stated retrieval and dilution conditions if available, then assess controls alongside the section (general IHC practice). No SEC63-specific retrieval, fixation sensitivity or dilution is established by the supplied UniProt and HPA records. |
| IF/ICC Q: What localization should an IF experiment check? | A: Check for ER localization; HPA reports a supported ER main location in ICC-IF, consistent with UniProt's ER membrane annotation (HPA subcellular; UniProt Q9UGP8). IF/ICC has its own guide; this IHC section supplies no IF protocol option. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known positive area is blank. | Pancreatic exocrine glandular cells are reported High, so a blank result needs investigation (HPA tissue IHC). | Confirm those cells are present; review the run's positive control, antibody preparation and detection steps before interpreting the section as negative (general IHC practice). |
| Signal is mostly nuclear. | That compartment conflicts with SEC63's ER membrane annotation and HPA's cytoplasmic IHC profile (UniProt Q9UGP8; HPA tissue IHC). | Compare with the expected cytoplasmic control and a no-primary control; review chromogen and counterstain before scoring (general IHC practice). |
| Oral mucosal squamous cells stain strongly. | HPA reports SEC63 as Not detected in those cells; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Verify the cell type and inspect no-primary and detection controls. Do not infer specificity from intensity alone (general IHC practice). |
| Color covers stroma and empty areas. | This lacks the cellular cytoplasmic distribution expected from the HPA profile (HPA tissue IHC). Excess background is a possible technical cause (general IHC practice). | Inspect the no-primary control and review blocking, washing and chromogen exposure using the validated IHC workflow (general IHC practice). |
| Liver hepatocytes stain weakly despite UniProt's high liver expression statement. | The sources describe different measures: UniProt reports high levels in liver, while HPA scores hepatocyte IHC staining Low (UniProt Q9UGP8 tissue specificity; HPA tissue IHC). | Use the named pancreatic or salivary glandular cells as the stronger IHC reference; record hepatocyte staining against HPA's cell-specific Low result (HPA tissue IHC). |
| Two antibodies give different cell patterns. | HPA lists Supported IHC status for HPA053295 and HPA060535, but that status does not establish agreement for every specimen or a separate catalog antibody (HPA antibodies). | Compare matched sections and controls, and prioritize staining that tracks the reported cytoplasmic pattern in the named positive cells (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing SEC63 staining
A04852-1 · IHCIHC analysis of SEC63 using anti-SEC63 antibody (A04852-1). SEC63 was detected in a paraffin-embedded section of human thyroid cancer tissue
A04852-1 · IHCIHC analysis of SEC63 using anti-SEC63 antibody (A04852-1). SEC63 was detected in a paraffin-embedded section of mouse kidney tissue. Heat m
A04852-1 · IFIF analysis of SEC63 using anti-SEC63 antibody (A04852-1). SEC63 was detected in an immunocytochemical section of SIHA cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000025796-SEC63/tissue/cerebral+cortex#imid_15700411
HPA · Salivary glandHuman Protein Atlas IHC, Salivary gland. Source: https://www.proteinatlas.org/ENSG00000025796-SEC63/tissue/salivary+gland#imid_15700343
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000025796-SEC63/tissue/colon#imid_15700321
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000025796-SEC63/tissue/liver#imid_15700330
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA053295; green = SEC63, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA053295; green = SEC63, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SEC63 IHC & IF
🧪Run pancreas first and expect staining in exocrine glandular cells (HPA: High); use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected). On the pancreas slide, morphologically distinct cells without DAB signal can provide an internal background reference if present, but the supplied HPA rows do not identify a validated negative pancreatic cell type.
Positive control tissue: Pancreas (Exocrine glandular cells, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SEC63 in HEK293, Hep-G2, U2OS, ASC52telo, NIH 3T3, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary); and SEC63 knockout material as a biological negative. Block endogenous peroxidase in pancreas sections before HRP/DAB detection (selected-SKU caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the A04852-1 paraffin-section caption does not state a fixative (selected-SKU caption). Heat retrieval in EDTA at pH 8.0 was used in that IHC example; its necessity has not been established (selected-SKU caption). Frozen sections or IF are not shown to be easier by the supplied evidence; HPA ICC-IF supports ER localization, while pancreatic tissue autofluorescence should be checked if IF is pursued (HPA: Endoplasmic reticulum supported; standard IF practice).
HPA tissue IHC evidence for SEC63
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High 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 |
|---|---|---|---|---|
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SEC63 IHC Tips
Troubleshoot SEC63 staining in paraffin sections by checking retrieval, cellular pattern, controls and scoring against its expected ER localisation (UniProt Q9UGP8; HPA subcellular).
Which retrieval conditions should I try first for weak SEC63 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04852-1). The documented thyroid cancer staining used this retrieval before overnight incubation with 2 μg/ml primary antibody at 4°C (datasheet A04852-1). If staining is weak, compare a carefully controlled change in heating duration with the documented condition while keeping section thickness, antibody concentration and detection constant (standard IHC practice). Include a positive reference section and assess whether signal strengthens in cell bodies without a parallel increase at tissue edges or in cell-free areas (HPA tissue IHC; standard IHC practice).
Could fixation explain inconsistent SEC63 staining between paraffin blocks?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state its fixative, and the supplied evidence does not establish how SEC63 responds to fixation (datasheet A04852-1). Record each block’s fixative and fixation duration, then compare sections processed with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A04852-1; standard IHC practice). Inconsistent fixation can alter antigen accessibility and tissue morphology in IHC generally, but that observation alone cannot establish a SEC63-specific effect (standard IHC practice). Judge any improvement alongside preserved morphology and the expected cytoplasmic pattern, using matched reference tissue across runs (HPA tissue IHC; standard IHC practice).
What staining pattern should count as plausible SEC63 localisation?
Look for cytoplasmic staining consistent with an ER membrane protein, rather than assigning SEC63 to nuclei solely because a nucleus looks dark (UniProt Q9UGP8 subcellular location; HPA tissue IHC). HPA reports general cytoplasmic expression in tissue and a supported ER location in cellular imaging (HPA tissue IHC; HPA subcellular). SEC63 has 3 transmembrane segments near its N terminus and a long cytoplasmic region spanning residues 210–760, so interpretation should focus on cellular distribution rather than an imagined soluble nuclear pool (UniProt Q9UGP8 topology). Compare the pattern with morphology, a reference section and a no-primary control before calling atypical deposits positive (standard IHC practice).
How should an unknown antibody epitope affect SEC63 troubleshooting?
Do not assume the catalog antibody binds a particular membrane-facing region unless its epitope is documented; the supplied caption gives no epitope coordinates (datasheet A04852-1). SEC63 has lumenal residues 91–188 and cytoplasmic residues 210–760, making epitope accessibility a plausible variable to test experimentally (UniProt Q9UGP8 topology; standard IHC practice). The record lists 0 isoforms, so this evidence does not support explaining discordant sections by a documented isoform switch (UniProt Q9UGP8 isoforms). Keep retrieval and detection fixed when comparing antibody conditions, and confirm that any stronger signal retains the expected cytoplasmic distribution (HPA tissue IHC; standard IHC practice).
How can I check SEC63 localisation by multiplex IF?
For an IF comparison, pair SEC63 with a separately validated marker for the expected cell type, such as exocrine glandular cells in pancreas, and inspect overlap at the cell level (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and favour a longer-wavelength channel for the weaker signal after measuring tissue autofluorescence in an unstained control (standard IF practice). Permeabilisation should be optimised against the antibody’s mapped epitope: SEC63 includes lumenal residues 91–188 and cytoplasmic residues 210–760, while this antibody’s epitope is unspecified (UniProt Q9UGP8 topology; datasheet A04852-1). Compare single-stain controls and expect an ER-associated intracellular pattern before interpreting apparent multiplex overlap (HPA subcellular; standard IF practice).
What should I change when DAB appears throughout the section?
First compare a no-primary section with the stained section; shared DAB deposits point toward detection or tissue background rather than antibody-dependent SEC63 signal (standard IHC practice). Include a peroxidase-blocking step for chromogenic HRP detection and verify that blocking, washes and DAB development are consistent across slides (standard IHC practice). The documented workflow used 10% goat serum, an anti-rabbit secondary for 30 minutes at 37°C, and DAB, but those details do not establish the cause of background in another specimen (datasheet A04852-1). If background persists, adjust primary concentration or development time in matched sections and preserve a reference control (standard IHC practice).
How should I quantify SEC63 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, and exclude folds, torn edges and necrotic areas by a prespecified rule (standard IHC practice). For cellular expression, record the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories and their cell percentages (standard IHC practice). Normalise counts to the number of evaluable cells; if measuring positive-cell density, report cells per mm² of viable tissue and retain the same area definition across sections (standard IHC practice). Use matched retrieval and DAB development, and compare like cell types because HPA reports differing SEC63 staining levels among tissues and cells (datasheet A04852-1; HPA tissue IHC).
How do I distinguish convincing SEC63 staining from artefact?
A convincing result follows cell morphology and shows a cytoplasmic pattern compatible with SEC63’s ER membrane location (HPA tissue IHC; UniProt Q9UGP8 subcellular location). For context, HPA reports high staining in pancreatic exocrine glandular cells and no detected staining in oral mucosal squamous epithelial cells; these are reference patterns, not absolute controls for every preparation (HPA tissue IHC). Treat isolated nuclear signal, edge-concentrated deposits, staining over necrosis, or DAB present in a no-primary control as reasons to investigate artefact (HPA subcellular; standard IHC practice). Review viable matched areas and verify peroxidase blocking before interpreting unexpected chromogenic signal as a change in SEC63 expression (standard IHC practice).
Boster reagents
Best SEC63 / Translocation protein SEC63 homolog IHC Antibodies
The catalog antibody has IHC images from paraffin sections of human thyroid cancer and mouse kidney, plus IF images from SIHA cells and human thyroid cancer sections (catalog image captions).
Real IHC data
Anti-SEC63 Antibody ®
Cat # A04852-1
A04852-1 has IHC images from paraffin sections of human thyroid cancer and mouse kidney (catalog IHC captions). A04852-1 also has IF images from SIHA cells and paraffin sections of human thyroid cancer (catalog IF captions).
Which to pick: Choose A04852-1 for paraffin section IHC: its own captions document both tissue examples, although the fixative is unreported (catalog IHC captions). Choose the same SKU for IF/ICC because those applications are listed and its IF captions document SIHA cells and human thyroid cancer sections (catalog applications; catalog IF captions). For work across species, the catalog lists human, mouse and rat reactivity, but its IHC dilution listing covers human and mouse only; rat IHC is unverified here (catalog reactivity; catalog dilution listing).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UGP8 (SEC63_HUMAN, Translocation protein SEC63 homolog).
- Human Protein Atlas. SEC63 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SEC63 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. SEC63 antibody validation summary (2 antibodies).
- Loss of heterozygosity is present in SEC63 germline carriers with polycystic liver disease. PloS one 2012 — PMC3508994.
- The uterine expression of SEC63 gene is up-regulated at implantation sites in association with the decidualization during the early pregnancy in mice. Reproductive biology and endocrinology : RB&E 2009 — PMC2655295.
- Activation of ACLY by SEC63 deploys metabolic reprogramming to facilitate hepatocellular carcinoma metastasis upon endoplasmic reticulum stress. Journal of experimental & clinical cancer research : CR 2023 — PMC10150531.
- Mutation of sec63 in zebrafish causes defects in myelinated axons and liver pathology. Disease models & mechanisms 2013 — PMC3529346.
- PubMed PMID:10543453 — UniProt-cited evidence.
- PubMed PMID:10799540 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SEC63 (Translocation protein SEC63 homolog). Expected localisation General cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04852-1). Validated IHC antibody: A04852-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Pancreas (Exocrine glandular cells); negative Oral mucosa. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A04852-1); 2. Fixation: Image fixative and duration unreported (datasheet A04852-1); 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 A04852-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04852-1); 8. Primary antibody: Rabbit anti-SEC63, 2-5 μg/ml (datasheet A04852-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04852-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A04852-1); verify before use.; Caveat: Hepatocytes stain low despite reported high liver expression (HPA tissue IHC; UniProt); Isoform / epitope: 0 isoforms; epitopes may face the ER lumen or cytoplasm (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q9UGP8, HPA tissue IHC · Supported, HPA ICC-IF · endoplasmic reticulum., Datasheet A04852-1, PMC3508994, PMC2655295. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SEC63/immunohistochemistry-ihc-guide.