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SLITRK4 / SLIT and NTRK-like protein 4 · IHC design guide
Design Immunohistochemistry for SLITRK4
Plan paraffin IHC for SLITRK4 using its reported cytoplasmic tissue staining and annotated membrane topology (HPA tissue IHC; UniProt). Compare cerebral cortical neurons with epididymal glandular cells, where staining is low and medium, respectively (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLITRK4 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 staining (HPA tissue IHC); cell membrane protein (UniProt) | |
| Staining pattern | Low cytoplasmic staining in cerebral cortical neurons (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Epididymis | |
| 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A14468) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Higher expression in some astrocytic brain tumors (UniProt) | |
| Isoform / epitope | No listed isoforms; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended SLITRK4 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published uterine leiomyosarcoma IHC workflow (PMC3965648).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A14468); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| 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-SLITRK4, 2.5 μg/mL (datasheet A14468) |
| 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 | SLITRK4-positive staining in glandular cells of epididymis (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues, including neuronal cells in cerebral cortex and cells in adrenal gland. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page antigen-retrieval specification).
Section 2
What Is the Expected SLITRK4 Staining Pattern?
SLITRK4 is a cell-membrane protein with an extracellular region at residues 19–618 and a cytoplasmic region at 640–837 (UniProt Q8IW52 topology). In tissue IHC, expect cytoplasmic staining in selected cells, including cortical neurons and adrenal gland cells, rather than uniform staining across a section (HPA tissue IHC). The HPA tissue profile is rated Enhanced, with medium consistency between staining and RNA data and external verification pending (HPA tissue IHC).
| Selective cytoplasmic staining in epididymal glandular cells, or weaker staining in cortical neurons. | This fits reported IHC: epididymal glandular cells are Medium; cortical neuronal cells are Low (HPA tissue IHC). Evaluate staining in identifiable cells against background on the same section (general IHC practice). |
| Predominantly nuclear staining, with no convincing cytoplasmic or membrane-associated component. | A nuclear-only pattern conflicts with the reported cytoplasmic tissue pattern and membrane assignment (HPA tissue IHC; UniProt Q8IW52 subcellular location). Treat it as suspect; review controls and detection before assigning it to SLITRK4 (general IHC practice). |
| Strong staining in adipocytes or appendix glandular cells, while expected cells are unstained. | Both cell populations are reported Not detected by HPA (HPA tissue IHC). Cross-reactivity or endogenous chromogen-producing activity is possible; the slide alone cannot distinguish them (general IHC practice). |
| Broad, hazy color over many cell types and tissue spaces. | That distribution obscures the selective pattern reported by HPA (HPA tissue IHC). Check reagent background, blocking and washes, and inspect a no-primary control before scoring cells (general IHC practice). |
| No signal in an epididymis section containing intact glandular cells. | That is discordant with HPA's Medium glandular-cell result, but one negative section does not prove absent protein (HPA tissue IHC; general IHC practice). Check section quality and controls, then optimize the IHC-P workflow. |
💡Expected SLITRK4 appearanceCall positive when identifiable epididymal glandular cells show Medium cytoplasmic staining, or cortical neurons show Low cytoplasmic staining (HPA tissue IHC); isolated nuclear signal or widespread color in reported negative cells is suspect (HPA tissue IHC; general IHC practice).
| Topology and compartment | SLITRK4 spans the membrane at residues 619–639, with extracellular and cytoplasmic regions (UniProt Q8IW52 topology). HPA reports cytoplasmic tissue IHC staining, so assess the observed cell pattern without requiring a crisp membrane rim in every positive cell (HPA tissue IHC). |
| Choice of reference tissue | Epididymal glandular cells provide a reported Medium IHC reference; cortical neurons, adrenal glandular cells and cerebellar molecular-layer neuropil are Low (HPA tissue IHC). A weak result in a Low reference needs careful background comparison (general IHC practice). |
| Antibody evidence | HPA lists HPA000431 as IHC Enhanced, while its overall tissue profile has medium staining–RNA consistency and awaits external verification (HPA antibody record; HPA tissue IHC). Interpret unexpected staining cautiously and retain the antibody and tissue controls in the record (general IHC practice). |
| IF/ICC: should a membrane pattern be expected? | UniProt assigns SLITRK4 to the cell membrane, and HPA summarizes its subcellular location as Membrane (UniProt Q8IW52; HPA subcellular). HPA provides no ICC/IF image cell lines or main-location detail here, and the listed antibody has no ICC validation entry (HPA subcellular; HPA antibody record). |
| Endogenous detection activity | Chromogenic IHC can show signal from endogenous tissue activity when the detection system is inadequately controlled (general IHC practice). Use an appropriate no-primary control to evaluate unexpected color; HPA's SLITRK4 staining levels do not identify its cause (HPA tissue IHC; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Epididymal glandular cells are blank. | The result disagrees with HPA's Medium reference; section quality, detection or assay settings may contribute (HPA tissue IHC; general IHC practice). | Confirm glandular morphology and control performance. Review the catalog antibody's IHC-P instructions, then optimize retrieval and antibody dilution within that workflow (general IHC practice). |
| Cortical neurons or adrenal glandular cells look negative. | Both are reported Low, so their expected staining may be difficult to distinguish from background (HPA tissue IHC; general IHC practice). | Compare with a no-primary control and an epididymal reference before calling absence; score only identifiable cells (HPA tissue IHC; general IHC practice). |
| Only nuclei are colored. | Nuclear-only staining is discordant with HPA's cytoplasmic tissue pattern and UniProt's membrane location (HPA tissue IHC; UniProt Q8IW52). | Check the no-primary control and detection reagents; repeat with the IHC-validated antibody before interpreting nuclear color as target staining (general IHC practice). |
| Reported negative cell types stain strongly. | Adipocytes and appendix glandular cells are Not detected in the supplied HPA profile; nonspecific antibody binding or endogenous detection activity could explain color (HPA tissue IHC; general IHC practice). | Confirm cell identity, inspect a no-primary control, and review blocking, washes and antibody dilution. Avoid scoring those cells as positive from color alone (general IHC practice). |
| Diffuse background masks cell boundaries. | Background prevents reliable comparison with HPA's selective cytoplasmic observations (HPA tissue IHC; general IHC practice). | Review washes, blocking and detection controls; adjust conditions under the catalog antibody's IHC-P workflow, then compare the repeated section with its no-primary control (general IHC practice). |
| A stained section differs from an HPA example. | HPA reports medium staining–RNA consistency and pending external verification, so a single reference image cannot settle an unexpected pattern (HPA tissue IHC). | Record the tissue, cell type, compartment and intensity; compare matched controls and repeat the IHC assessment before making a biological claim (general IHC practice). |
Experiment evidence
IHC & IF images showing SLITRK4 staining
A14468 · IHCImmunohistochemistry of Slitrk4 in human brain tissue with Slitrk4 antibody at 2.5 μg/mL.
A14468 · IFImmunofluorescence of slitrk4 in human brain tissue with slitrk4 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000179542-SLITRK4/tissue/cerebral+cortex#imid_227586
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000179542-SLITRK4/tissue/cerebellum#imid_227597
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000179542-SLITRK4/tissue/colon#imid_227512
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000179542-SLITRK4/tissue/liver#imid_227529
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLITRK4 IHC & IF
🧪Run epididymis first and look for staining in glandular cells (HPA: Medium); use appendix glandular cells as a negative tissue comparison (HPA: Not detected). The supplied HPA rows do not identify validated negative cells within epididymis, so treat neighboring non-glandular cells as a background reference only, without assuming they lack SLITRK4 (HPA: epididymis glandular cells Medium).
Positive control tissue: Epididymis (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for SLITRK4; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary or secondary-only and host-species- and clonality-matched isotype controls; use SLITRK4 knockout tissue, if available, or an immunizing-peptide block as a biological specificity control. For chromogenic IHC, quench endogenous peroxidase and check the epididymis control slide for residual background; block endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A14468 brain-tissue IHC caption also leaves the fixative unreported (A14468 caption). The evidence does not establish whether frozen sections or IF are easier than paraffin-section IHC for SLITRK4 (supplied application evidence). Check epididymal glandular staining against the no-primary control for tissue background (HPA: epididymis glandular cells Medium; standard IHC practice).
HPA tissue IHC evidence for SLITRK4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLITRK4 IHC Tips
Troubleshoot SLITRK4 staining in paraffin sections by checking retrieval, tissue controls, cellular distribution and scoring before interpreting chromogenic signal.
How should I optimize retrieval when SLITRK4 staining is weak?
Start with citrate pH 6.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool in the retrieval solution, then compare a matched human brain section with a no-primary control under identical detection conditions (selected A14468 caption; standard IHC practice). If staining remains weak, test a longer retrieval time or an alkaline buffer as a fallback, changing only one variable per run (standard IHC practice). Excess retrieval can damage morphology or increase nonspecific staining, so judge signal together with intact tissue architecture (standard IHC practice). Record the buffer, temperature and time used for every scoring batch (standard IHC practice).
Could fixation explain weak or uneven SLITRK4 staining?
Target-specific fixation sensitivity is unknown: the selected human brain IHC caption reports 2.5 µg/mL antibody but does not state a fixative (selected A14468 caption). For paraffin sections, document the actual fixative, fixation duration and processing history, and compare sections processed together before changing antibody concentration (standard IHC practice). If tissue is available, use consistently processed control material to distinguish a processing problem from a staining-run problem (standard IHC practice). Avoid attributing weak signal to SLITRK4 topology or glycosylation, because those annotations do not establish fixation sensitivity (UniProt Q8IW52 topology and glycosylation). Treat differences between tissue blocks cautiously when their fixation histories are unavailable (standard IHC practice).
What staining pattern is plausible for SLITRK4 in brain sections?
SLITRK4 is annotated at the cell membrane, with an extracellular region at residues 19–618 and a cytoplasmic region at 640–837 (UniProt Q8IW52 topology). HPA reports cytoplasmic tissue staining, including neuronal cells in cerebral cortex, so assess both cell-associated membrane accentuation and cytoplasmic signal against controls (HPA tissue IHC). A diffuse field-wide precipitate without cellular boundaries is less persuasive than staining tied to identifiable cells and preserved morphology (standard IHC practice). Check neuronal areas against a matched no-primary section, and document whether the signal lies in cell bodies, processes or surrounding tissue (HPA tissue IHC; standard IHC practice). Do not assign a subcellular epitope from the staining pattern alone (standard IHC practice).
How can epitope location change the IHC result?
The supplied record lists 0 isoforms, so an isoform-specific explanation for discordant staining is unsupported here (UniProt Q8IW52 isoforms). SLITRK4 has an extracellular region at residues 19–618 and a cytoplasmic region at 640–837, separated by a transmembrane segment (UniProt Q8IW52 topology). Its annotated glycosylation sites at residues 81 and 325 lie in the extracellular region, but the catalog antibody’s epitope is not supplied (UniProt Q8IW52 topology and glycosylation; selected A14468 caption). Consult the antibody’s documented immunogen before interpreting an accessibility problem, and compare retrieval conditions on matched sections (standard IHC practice). Without epitope mapping, staining cannot identify which side of the membrane the antibody recognized (standard IHC practice).
How should I assess SLITRK4 in a multiplex IF experiment?
For the separate IF/ICC workflow, pair SLITRK4 with a validated neuronal marker when assessing cerebral cortex, and include single-stain controls to check channel overlap (HPA tissue IHC; standard IF practice). Favor a far-red fluorophore for a weak SLITRK4 signal if the tissue shows strong shorter-wavelength autofluorescence, and inspect an unstained section first (standard IF practice). If the antibody recognizes the cytoplasmic region at residues 640–837, permeabilization is needed for access in fixed cells; an extracellular epitope may be accessible without it (UniProt Q8IW52 topology; standard IF practice). Because the catalog epitope is unspecified, compare permeabilized and nonpermeabilized preparations before assigning compartment-specific signal (selected A14468 caption; standard IF practice). IHC staining at 2.5 µg/mL does not establish an IF dilution (selected A14468 caption).
How can I reduce diffuse brown staining without losing SLITRK4 signal?
First compare the stained section with a no-primary control and inspect whether brown signal follows cells or spreads across tissue and section edges (standard IHC practice). Block endogenous peroxidase before chromogenic detection, apply an appropriate protein block, and optimize washes and antibody concentration one variable at a time (standard IHC practice). The selected human brain IHC caption reports 2.5 µg/mL for A14468, providing a reference point for that image rather than a guaranteed optimum for every run (selected A14468 caption). Shorten DAB development if the no-primary control is clean but dense precipitate obscures cellular detail (standard IHC practice). Recheck retrieval-induced tissue damage when background rises after hotter or longer heating (standard IHC practice).
What should I score when comparing SLITRK4 across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region and cell population before scoring, then report an H-score, percentage of positive cells, or positive-cell density per mm² (standard IHC practice). Normalize cell counts to the number of evaluable cells and density to the measured tissue area; exclude folds, tears and necrotic regions consistently (standard IHC practice). Record staining intensity and subcellular distribution separately, because HPA describes cytoplasmic tissue staining while UniProt annotates a membrane protein (HPA tissue IHC; UniProt Q8IW52 subcellular location). Use identical retrieval, detection, counterstain and image-analysis thresholds within a comparison set (standard IHC practice). Include positive and no-primary controls in each run before comparing scores (standard IHC practice).
When is a positive SLITRK4 result credible rather than artefactual?
A credible result has cell-associated staining, preserved morphology and an appropriate control pattern; the selected antibody has a human brain IHC image at 2.5 µg/mL (selected A14468 caption; standard IHC practice). Neuronal staining in cerebral cortex is plausible, although HPA rates that signal low and describes cytoplasmic expression (HPA tissue IHC). Strong staining restricted to section edges, necrotic tissue or a no-primary control warrants investigation of processing, background or endogenous enzyme activity (standard IHC practice). Staining in an unexpected compartment should be checked against SLITRK4’s membrane annotation and the antibody’s known epitope, if available (UniProt Q8IW52 subcellular location; standard IHC practice). HPA labels its tissue evidence Enhanced while noting only medium staining–RNA consistency and pending external verification (HPA tissue IHC).
Boster reagents
Best SLITRK4 / SLIT and NTRK-like protein 4 IHC Antibodies
A14468 has IHC data from human brain tissue at 2.5 μg/mL and IF data from human brain tissue at 20 μg/mL (catalog image captions); the catalog lists Human, Mouse and Rat reactivity (catalog: A14468).
Real IHC data
Anti-SLIT and NTRK-like protein 4 Slitrk4 Antibody
Cat # A14468
A14468 is listed for IHC-P and IF (catalog: A14468 applications). Its IHC and IF images show human brain tissue at 2.5 μg/mL and 20 μg/mL, respectively (catalog: A14468 image captions).
Which to pick: Choose A14468 for paraffin-section IHC: IHC-P is listed, and its own IHC caption shows human brain tissue at 2.5 μg/mL (catalog: A14468 applications and IHC image caption). For tissue IF, its own IF caption shows human brain tissue at 20 μg/mL; ICC is not listed as a validated application (catalog: A14468 IF image caption and applications). Human, Mouse and Rat reactivity is listed for A14468, but the supplied IHC and IF images show human tissue only; the IHC fixative is unreported (catalog: A14468 reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IW52 (SLIK4_HUMAN, SLIT and NTRK-like protein 4).
- Human Protein Atlas. SLITRK4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SLITRK4 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. SLITRK4 antibody validation summary (1 antibodies).
- Gene expression signatures of primary and metastatic uterine leiomyosarcoma. Human pathology 2014 — PMC3965648.
- PubMed PMID:15772651 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLITRK4 (SLIT and NTRK-like protein 4). Expected localisation Cytoplasmic staining (HPA tissue IHC); cell membrane protein (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A14468 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A14468); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 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-SLITRK4, 2.5 μg/mL (datasheet A14468); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A14468); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No listed isoforms; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Higher expression in some astrocytic brain tumors (UniProt). Evidence: UniProt Q8IW52, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A14468, PMC3965648. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLITRK4/immunohistochemistry-ihc-guide.