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SEZ6L2 / Seizure 6-like protein 2 · IHC design guide
Design Immunohistochemistry for SEZ6L2
Plan SEZ6L2 paraffin IHC around cytoplasmic staining in CNS neurons (HPA tissue IHC). This guide covers fixation handling, chromogenic detection, and scoring by cell type and compartment (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SEZ6L2 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 | Predominantly cytoplasmic in tissue (HPA tissue IHC); membrane and ER localisation (UniProt) | |
| Staining pattern | Cytoplasmic staining, mainly in CNS neurons (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11420-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11420-1) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is unreported (UniProt) | |
| Isoform / epitope | 6 isoforms; extracellular versus cytoplasmic epitope location matters (UniProt) |
Section 1
Recommended SEZ6L2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A11420-1) is paired with a published SEZ6L2 protocol for colorectal cancer tissue and xenograft sections (PMC7171412).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A11420-1) |
| Fixation | Image fixative and duration unreported (datasheet A11420-1); 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 A11420-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11420-1) |
| Primary antibody | Rabbit anti-SEZ6L2, 2-5 μg/ml (datasheet A11420-1) |
| Primary incubation | Overnight at 4 °C (datasheet A11420-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11420-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SEZ6L2-positive staining in ciliated cells (cell body) of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, mainly in CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A11420-1); the published CRC excerpt does not specify retrieval (PMC7171412).
Section 2
What Is the Expected SEZ6L2 Staining Pattern?
SEZ6L2 is annotated at the cell and endoplasmic reticulum membranes, with an extracellular region spanning residues 28–844 and a cytoplasmic tail at 866–910 (UniProt Q6UXD5 topology). In tissue IHC, expect mainly cytoplasmic staining in CNS neurons, with strong staining also reported in selected ciliated epithelia (HPA: tissue IHC profile; High in caudate neurons, bronchial ciliated cells and fallopian tube ciliated cells). HPA rates the tissue pattern Enhanced, with medium consistency against RNA expression (HPA: reliability).
| Strong staining in caudate or cerebral cortex neuronal cells, or in cerebellar molecular layer cells. | This fits the reported CNS pattern; cerebellar molecular layer staining is described as cytoplasmic/membranous (HPA: High in caudate and cerebral cortex neurons; High in cerebellar molecular layer cells). Judge the cell type and distribution together, rather than treating any isolated dark deposit as a positive cell (general IHC practice). |
| Staining concentrates in bronchial ciliated cell bodies or at fallopian tube ciliary tips. | Both are reported strong patterns, but their recorded positions differ (HPA: High in bronchial ciliated cell bodies; High at fallopian tube ciliary tips). A visible signal confined to those positions supports interpretation more strongly than widespread epithelial staining (general IHC practice). |
| A predominantly nuclear pattern appears where a membrane or cytoplasmic pattern is expected. | Nuclear localisation is absent from the supplied localisation records (UniProt Q6UXD5 subcellular location; HPA: cytoplasmic tissue profile). Treat it as discordant; check the counterstain, detection controls and antibody conditions before assigning SEZ6L2 positivity (general IHC practice). |
| Strong signal appears in an HPA-negative cell type, or colour covers tissue without clear cell boundaries. | For example, HPA reports no detection in adipocytes or lung alveolar cells (HPA: Not detected in adipocytes and alveolar cells). Cross-reactivity or endogenous detection activity may explain cell-specific unexpected signal; diffuse colour may reflect background (general IHC practice). Neither appearance alone identifies its cause. |
| A known-positive region has little or no visible stain. | Check that the section contains the specified cells: HPA reports High staining in caudate neurons and bronchial ciliated cells (HPA: tissue IHC). If they are present, review antibody, retrieval and detection controls before interpreting the absence as biology (general IHC practice). HPA's Enhanced rating does not guarantee every run will produce signal (HPA: reliability). |
💡Expected SEZ6L2 appearanceCall a section positive when appropriately placed cytoplasmic or membrane-associated signal is clear in expected cells, especially High-staining CNS neurons or selected ciliated cells (HPA: tissue IHC; UniProt Q6UXD5 location); diffuse colour, nuclear-only staining or strong signal in HPA-negative cells warrants control review (general IHC practice; HPA: negative tissue entries).
| Membrane topology and compartment | SEZ6L2 has one transmembrane segment at 845–865 and an extracellular region at 28–844 (UniProt Q6UXD5 topology). Tissue IHC may nevertheless appear mainly cytoplasmic (HPA: tissue IHC profile); do not require a crisp surface outline in every positive cell. |
| Tissue and cell selection | HPA records High staining in CNS neurons and selected ciliated cells, Medium in several glandular populations, and Low in kidney tubule cells (HPA: tissue IHC). Match comparisons to the annotated cell population; a whole-organ label alone can hide the relevant staining pattern. |
| Evidence strength | The tissue pattern is rated Enhanced, with medium consistency between antibody staining and RNA expression (HPA: reliability). HPA lists one rabbit polyclonal antibody, HPA064471, as IHC Enhanced; that rating supports the reported pattern but does not validate every unexpected compartment (HPA: antibodies). |
| Isoforms, processing and epitope | UniProt lists 6 isoforms, a signal peptide at 1–27, and a chain at 28–910 (UniProt Q6UXD5). No antibody epitope is supplied here, so these annotations cannot establish which isoforms the IHC antibody detects or predict its staining after processing. |
| Retrieval conditions | Retrieval is a condition to record and optimise with controls in paraffin-section IHC (general IHC practice). Target-specific fixation sensitivity and an optimal SEZ6L2 retrieval condition are unreported in the supplied UniProt and HPA records. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in the expected positive control. | The sampled section may lack the annotated cells, or an IHC step may have failed (HPA: High in caudate neurons and bronchial ciliated cells; general IHC practice). | Confirm the relevant cells on the counterstain, then inspect retrieval, primary antibody and chromogenic detection controls on a known-positive section (general IHC practice). Do not infer loss of SEZ6L2 from a failed control. |
| Strong colour appears in adipocytes or lung alveolar cells. | Those cells are reported as Not detected by HPA; unexpected signal may reflect cross-reactivity or endogenous detection activity (HPA: tissue IHC; general IHC practice). | Compare the expected positive cells, use a primary-omission control and check the detection-system block appropriate to the chromogen (general IHC practice). Investigate persistent cell-specific staining before scoring it as SEZ6L2. |
| Colour is diffuse across tissue or obscures cell boundaries. | Nonspecific background, excessive primary antibody or detection signal are possible technical causes (general IHC practice). The supplied HPA profile describes cellular staining rather than uniform colour (HPA: tissue IHC profile). | Review the primary-omission control, blocking and washes; titrate the antibody and detection time against a known-positive section (general IHC practice). Retain conditions that preserve interpretable cell localisation. |
| The strongest signal looks nuclear. | That compartment conflicts with the supplied membrane and cytoplasmic annotations (UniProt Q6UXD5 subcellular location; HPA: tissue IHC profile). Counterstain overlap or nonspecific signal could mislead interpretation (general IHC practice). | Inspect chromogen and counterstain separately where possible, compare controls, and score cytoplasmic or membrane-associated staining in the expected cells (general IHC practice; HPA: tissue IHC). |
| Only weak signal is seen in a candidate control tissue. | Some listed cells are inherently Low or Medium in the HPA observations; kidney tubule cells are Low, whereas caudate neurons are High (HPA: tissue IHC). | Choose a section containing an HPA High cell population to assess the staining run, and compare the same cell type across sections (HPA: tissue IHC; general IHC practice). Avoid calling a Low population a failed control solely because its signal is faint. |
| Can the tissue IHC pattern define the expected ICC-IF result? | HPA's subcellular summary says Membrane but gives no main location or cell lines with ICC-IF images; HPA064471 has no ICC status in the supplied list (HPA: subcellular record; HPA: antibodies). | Use membrane localisation as a hypothesis and establish ICC-IF localisation with suitable cellular controls on its own guide page (UniProt Q6UXD5 location; general IF practice). Do not treat tissue IHC intensity as an ICC-IF validation result. |
Experiment evidence
IHC & IF images showing SEZ6L2 staining
A11420-1 · IHCIHC analysis of SEZ6L2 using anti-SEZ6L2 antibody (A11420-1). SEZ6L2 was detected in a paraffin-embedded section of mouse brain tissue. Heat
A11420-1 · IHCIHC analysis of SEZ6L2 using anti-SEZ6L2 antibody (A11420-1). SEZ6L2 was detected in a paraffin-embedded section of rat brain tissue. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000174938-SEZ6L2/tissue/cerebral+cortex#imid_18265239
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000174938-SEZ6L2/tissue/colon#imid_18265165
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000174938-SEZ6L2/tissue/liver#imid_18265166
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000174938-SEZ6L2/tissue/pancreas#imid_18265117
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SEZ6L2 IHC & IF
🧪Run bronchus first and require staining in ciliated cell bodies (HPA: High in bronchial ciliated cells). Run adipose tissue as a negative comparator, assessing adipocytes (HPA: Not detected in adipocytes); on the bronchus slide, luminal debris and areas without cells should remain at background, while other cell types should not be presumed negative.
Positive control tissue: Bronchus (Ciliated cells (cell body), 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 SEZ6L2; derive a cell-line control from the positive tissue's cell type (Ciliated cells (cell body)) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality where applicable (selected-SKU caption: rabbit primary antibody). Use SEZ6L2 knockout tissue if available, or peptide competition with the antibody’s immunizing peptide; quench endogenous peroxidase and check background before interpreting HRP–DAB staining (selected-SKU caption: HRP–DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not state the fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0 on a paraffin-embedded mouse brain section, but retrieval dependence is unreported; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU caption: paraffin section and EDTA retrieval). In bronchus, assess the ciliated cell body separately from luminal debris that could obscure the epithelial signal (HPA: High in bronchial ciliated cell bodies).
HPA tissue IHC evidence for SEZ6L2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bronchus | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Molecular layer cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (tip of cilia) | 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 → |
| Duodenum | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SEZ6L2 IHC Tips
Troubleshoot SEZ6L2 chromogenic IHC by checking retrieval, tissue morphology, cell identity and staining compartment against the available evidence.
Which retrieval conditions should I use when SEZ6L2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A11420-1). The selected mouse-brain image used that retrieval, followed by 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption A11420-1). If staining remains weak, compare heating durations within your instrument’s validated range while holding section thickness, antibody concentration and detection constant (standard IHC practice). Check whether neuronal staining improves without loss of tissue detail or a rise in diffuse DAB signal (HPA: high in caudate neuronal cells; standard IHC practice). Treat a different buffer or pH as a fallback requiring a matched comparison (datasheet A11420-1; standard IHC practice).
How can I troubleshoot fixation when SEZ6L2 staining varies between paraffin blocks?
Target-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (caption A11420-1). Record the fixative, time in fixative and processing history for each block, then compare sections using the same EDTA pH 8.0 retrieval and detection run (datasheet A11420-1; standard IHC practice). Assess nuclear detail, tissue integrity and staining together before attributing a weak result to fixation (standard IHC practice). Include a consistently staining control block in each run, such as a previously verified neuronal specimen, to distinguish block variation from run variation (HPA: high in caudate neuronal cells; standard IHC practice).
Should SEZ6L2 appear at the membrane or within the cytoplasm in IHC?
SEZ6L2 is annotated at the cell membrane and endoplasmic-reticulum membrane, with an extracellular region spanning residues 28–844 and a cytoplasmic tail at 866–910 (UniProt Q6UXD5 topology and subcellular annotation). HPA reports predominantly cytoplasmic tissue staining, mainly in the CNS, and high staining in cerebellar molecular-layer cells at the cytoplasm or membrane (HPA tissue IHC). Evaluate cell-associated membrane and cytoplasmic DAB staining in the expected cells, while recognizing that chromogenic resolution may not separate adjacent compartments (UniProt Q6UXD5; HPA tissue IHC; standard IHC practice). A ciliary pattern can also be relevant: HPA reports high signal at fallopian-tube cilia tips (HPA tissue IHC).
Could SEZ6L2 isoforms or epitope position explain inconsistent staining?
SEZ6L2 has 6 annotated isoforms, but the supplied antibody evidence does not map this antibody’s epitope or establish which isoforms it recognizes (UniProt Q6UXD5 isoforms; caption A11420-1). Its extracellular region spans residues 28–844, whereas residues 866–910 form the cytoplasmic tail, so an epitope map is needed before assigning staining to either side (UniProt Q6UXD5 topology). The protein also has 9 annotated glycosylation sites and repeated CUB and Sushi domains; these features identify plausible epitope contexts, without establishing their effect on this assay (UniProt Q6UXD5). Compare independent antibodies with documented, nonoverlapping epitopes on matched sections if isoform coverage or epitope accessibility remains in doubt (standard IHC practice).
How should I use IF to examine a questionable SEZ6L2 IHC pattern?
Use IF as a separate spatial check alongside chromogenic IHC; the supplied catalog example documents paraffin-section IHC, and HPA lists no ICC/IF images here (caption A11420-1; HPA subcellular). Multiplex SEZ6L2 with a marker for the expected neuronal or ciliated cell population, and choose a red or far-red channel after checking tissue autofluorescence and single-stain controls (HPA tissue IHC; standard IF practice). On intact cells, keep the membrane unpermeabilized when testing access to an extracellular epitope, but permeabilize when testing an intracellular tail or endoplasmic-reticulum epitope (UniProt Q6UXD5 topology; standard IF practice). Because this antibody’s epitope is unspecified, compare those conditions before interpreting a missing IF signal as biological absence (caption A11420-1; standard IF practice).
What controls help distinguish SEZ6L2 signal from diffuse brown background?
Begin with the documented 10% goat-serum block and 2 μg/ml primary concentration as assay reference points (caption A11420-1). Include a no-primary control, block endogenous peroxidase before HRP detection, and shorten DAB development if the control accumulates brown signal (standard IHC practice). The selected image used a peroxidase-conjugated secondary for 30 minutes at 37°C and DAB as chromogen; those settings describe that example, rather than proving specificity in another block (caption A11420-1). Compare background in expected positive neuronal cells with areas where HPA reports no detected signal, such as adipocytes, while accounting for differences in tissue composition (HPA tissue IHC; standard IHC practice).
How should I score SEZ6L2 IHC across samples with different cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region of interest and relevant cell population before scoring, then report the percentage of positive cells and an intensity-based H-score on the conventional 0–300 scale (standard IHC practice). For spatial questions, report positive-cell density per mm² of viable tissue and normalize counts to the area or number of eligible cells examined (standard IHC practice). Score neuronal and ciliated populations separately because HPA reports high staining in caudate neuronal cells, bronchial ciliated-cell bodies and fallopian-tube cilia tips (HPA tissue IHC). Keep retrieval, DAB development, imaging and positivity thresholds consistent across samples, and record excluded damaged or necrotic areas (datasheet A11420-1; standard IHC practice).
Which patterns support genuine SEZ6L2 staining rather than an IHC artefact?
Give greatest weight to reproducible cell-associated staining in plausible compartments: SEZ6L2 is annotated at cell and endoplasmic-reticulum membranes, and HPA reports predominantly cytoplasmic tissue staining (UniProt Q6UXD5; HPA tissue IHC). High staining in caudate or cerebral-cortex neuronal cells, bronchial ciliated cells or fallopian-tube cilia tips has tissue-IHC precedent (HPA tissue IHC). A diffuse nuclear pattern needs particular scrutiny because the supplied localisation evidence identifies membrane-associated locations (UniProt Q6UXD5; HPA subcellular). Check whether staining follows section edges, damaged or necrotic areas, or persists in a no-primary peroxidase control before calling it SEZ6L2-positive (standard IHC practice).
Boster reagents
Best SEZ6L2 / Seizure 6-like protein 2 IHC Antibodies
The IHC-validated anti-SEZ6L2 antibody has paraffin-section images from mouse and rat brain and human testis (catalog: IHC image captions). No IF/ICC images are supplied (catalog: IF images).
Real IHC data
Anti-SEZ6L2 Antibody ®
Cat # A11420-1
A11420-1 is listed for IHC and reacts with human, mouse and rat samples (catalog: applications and reactivity). Its IHC captions show paraffin sections of mouse and rat brain and human testis using 2 μg/ml primary antibody (catalog: IHC image captions).
Which to pick: Choose A11420-1 for paraffin-section tissue IHC; its mouse brain image used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (A11420-1 IHC caption). For work across species, it lists human, mouse and rat reactivity, with IHC images from each species; the fixative is unreported (catalog: reactivity; A11420-1 IHC captions). No IF/ICC application or image is supplied for A11420-1, so there is no IF/ICC-validated SKU to recommend from this catalog (catalog: applications and IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6UXD5 (SE6L2_HUMAN, Seizure 6-like protein 2).
- Human Protein Atlas. SEZ6L2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SEZ6L2 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SEZ6L2 antibody validation summary (1 antibodies).
- Overexpression of SEZ6L2 predicts poor prognosis in patients with cholangiocarcinoma. Translational cancer research 2020 — PMC8798643.
- SEZ6L2 knockdown impairs tumour growth by promoting caspase-dependent apoptosis in colorectal cancer. Journal of cellular and molecular medicine 2020 — PMC7171412.
- Sez6L2 autoimmunity induces cerebellar ataxia in mice. Journal of neuroinflammation 2025 — PMC12750838.
- Seizure-related 6 homolog like 2 autoimmunity: Neurologic syndrome and antibody effects. Neurology(R) neuroimmunology & neuroinflammation 2021 — PMC7641326.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SEZ6L2 (Seizure 6-like protein 2). Expected localisation Predominantly cytoplasmic in tissue (HPA tissue IHC); membrane and ER localisation (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11420-1). Validated IHC antibody: A11420-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (cell body)); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A11420-1); 2. Fixation: Image fixative and duration unreported (datasheet A11420-1); 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 A11420-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11420-1); 8. Primary antibody: Rabbit anti-SEZ6L2, 2-5 μg/ml (datasheet A11420-1); 9. Primary incubation: Overnight at 4 °C (datasheet A11420-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11420-1); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 6 isoforms; extracellular versus cytoplasmic epitope location matters (UniProt); Regulation: Expression regulation is unreported (UniProt). Evidence: UniProt Q6UXD5, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A11420-1, PMC7171412. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SEZ6L2/immunohistochemistry-ihc-guide.