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NLGN2 / Neuroligin-2 · IHC design guide
Design Immunohistochemistry for NLGN2
Plan NLGN2 paraffin IHC around cortical neuropil and cerebellar cytoplasm/membrane staining (HPA tissue IHC). This guide covers fixation, antigen retrieval, controls and chromogenic scoring, with 5 μg/mL as the catalog antibody’s IHC starting concentration (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NLGN2 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 | Cortical neuropil; cerebellar cytoplasm/membrane (HPA tissue IHC) | |
| Staining pattern | Cortical neuropil; enterocyte microvilli; Sertoli cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+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 A06595) | |
| Caveat | Staining–RNA consistency is medium (HPA tissue IHC) | |
| Regulation | Brain exceeds islet beta cells (UniProt) | |
| Isoform / epitope | No isoforms annotated; extracellular vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended NLGN2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is supplemented by published NLGN2 staining methods for breast tumor sections and rat hippocampus (PMC8595097; PMC3579928).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A06595); 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-NLGN2, 5 μg/mL (datasheet A06595) |
| 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 | NLGN2-positive staining in molecular layer cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: Medium). HPA tissue profile: Selective expression in CNS. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min for paraffin sections (page antigen-retrieval guidance); the rat free-floating sections used a separate workflow (PMC3579928).
Section 2
What Is the Expected NLGN2 Staining Pattern?
NLGN2 is a transmembrane protein with an extracellular region and cytoplasmic tail; brain localization includes postsynaptic membranes, while retinal localization includes presynaptic membranes (UniProt Q8NFZ4 topology and subcellular location). In paraffin IHC, expect staining in cortical neuropil and cerebellar molecular layer cells, with additional reported staining in enterocyte microvilli and Sertoli cells (HPA tissue IHC). HPA rates its tissue IHC reliability Enhanced, with medium consistency between staining and RNA data (HPA tissue IHC).
| Cortical neuropil stains at medium intensity; cerebellar molecular layer cells show cytoplasmic or membrane staining (HPA tissue IHC). | This matches the reported brain IHC pattern (HPA tissue IHC). Evaluate the stained structures in their tissue context: NLGN2 is associated with neuronal membranes and synapses, but the HPA cerebellar description also includes cytoplasm (UniProt Q8NFZ4 subcellular location; HPA tissue IHC). |
| Signal is predominantly nuclear, or an organelle-like pattern replaces the expected neuropil and membrane-associated staining (HPA tissue IHC; UniProt Q8NFZ4 subcellular location). | Treat this as discordant with the supplied IHC expectation and investigate specificity or staining artefact (HPA tissue IHC; general IHC practice). HPA separately reports mitochondrial ICC-IF staining; that observation alone does not establish the expected chromogenic IHC pattern (HPA subcellular ICC-IF; HPA tissue IHC). |
| Adipocytes or adrenal glandular cells stain strongly (HPA tissue IHC: both cell types not detected). | These are discordant findings in the named cell types, so assess antibody cross-reactivity and endogenous chromogenic detection activity (HPA tissue IHC; general IHC practice). Avoid treating an entire tissue as negative solely because HPA lists one cell type as not detected (HPA tissue IHC). |
| Colour spreads across tissue compartments and the surrounding slide without a clear cellular pattern (general IHC practice). | Diffuse background cannot establish NLGN2 localization. Review blocking, antibody concentration, washing and detection controls before scoring specific staining (general IHC practice); compare any retained pattern with the cell-level HPA observations (HPA tissue IHC). |
| A well-preserved cerebral cortex or cerebellum section has no detectable target signal (HPA tissue IHC: medium staining in the specified structures). | A missing positive-control pattern leaves this IHC run uninterpretable for negative samples (general IHC practice). Check tissue sampling and assay performance first; absence in one section does not overturn the reported tissue pattern (HPA tissue IHC; general IHC practice). |
💡Expected NLGN2 appearanceA convincing positive shows medium cortical neuropil or cerebellar molecular layer cell staining in the reported compartments, while strong nuclear staining or colour in HPA-not-detected cell types warrants investigation (HPA tissue IHC; UniProt Q8NFZ4 subcellular location).
| Membrane topology | NLGN2 spans residues 678–698, with extracellular residues 15–677 and cytoplasmic residues 699–835 (UniProt Q8NFZ4 topology). Interpret a membrane-associated pattern in light of this architecture; the supplied record does not identify the catalog antibody's epitope. |
| Choice of comparison tissue and cell | HPA reports medium staining in cortical neuropil, cerebellar molecular layer cells, enterocyte microvilli and Sertoli cells, but low staining in hippocampal and caudate neuronal cells (HPA tissue IHC). Compare the specified cells and structures, since a faint hippocampal result alone is a weak failure criterion. |
| Antibody-specific evidence | HPA lists HPA069278 as IHC Enhanced and HPA055321 as ICC Approved, without assigning both statuses to one antibody (HPA antibodies). HPA describes tissue IHC reliability as Enhanced with medium staining–RNA consistency (HPA tissue IHC); check which evidence applies to the antibody actually used. |
| IF/ICC Q: Does HPA mitochondrial staining define the expected IHC result? | A: No. HPA reports approved mitochondrial localization in ICC-IF, while its tissue IHC observations describe neuropil, cytoplasmic or membrane, and microvillar patterns (HPA subcellular ICC-IF; HPA tissue IHC). The listed ICC and IHC validation statuses belong to different antibodies (HPA antibodies). |
| Epitope and fixation limits | The supplied topology identifies extracellular and cytoplasmic regions, but no catalog-antibody epitope or target-specific fixation sensitivity is supplied (UniProt Q8NFZ4 topology; supplied evidence scope). Select antigen retrieval from the IHC-validated antibody's protocol and assess it with controls (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain appears in cortical neuropil or cerebellar molecular layer cells (HPA tissue IHC). | The expected positive-control pattern is absent; tissue sampling or assay performance may explain the result (HPA tissue IHC; general IHC practice). | Confirm the relevant structures are present, then review the catalog antibody's IHC-P protocol, retrieval, dilution and detection controls before interpreting test sections (general IHC practice). |
| The hippocampus is faint while cortex shows a clear neuropil pattern (HPA tissue IHC). | HPA reports low staining in hippocampal neuronal cells and medium staining in cortical neuropil (HPA tissue IHC). | Score each region against its own reported cell-level pattern; retain cortex as the stronger tissue comparison and avoid calling faint hippocampal staining an assay failure by itself (HPA tissue IHC). |
| Adipocytes or adrenal glandular cells show conspicuous chromogenic colour (HPA tissue IHC: not detected). | Cross-reactivity, nonspecific binding or endogenous detection activity are possible explanations (general IHC practice); the observed cell types conflict with the HPA entries (HPA tissue IHC). | Inspect reagent-omission and detection controls, review blocking, and compare staining in the named cells with a reported positive structure (general IHC practice; HPA tissue IHC). |
| Diffuse colour obscures neuropil and cell borders (general IHC practice). | Excess antibody or detection background can reduce contrast between specific staining and surrounding tissue (general IHC practice). | Review the antibody's IHC-P dilution guidance, blocking, washing and detection exposure; accept a result only when the reported cell-level distribution can be judged (general IHC practice; HPA tissue IHC). |
| A mitochondrial-looking pattern dominates the chromogenic section (HPA subcellular ICC-IF; HPA tissue IHC). | HPA's mitochondrial assignment comes from ICC-IF, whereas its tissue IHC evidence describes other structures and uses separately listed antibody validation (HPA subcellular ICC-IF; HPA tissue IHC; HPA antibodies). | Check antibody identity and IHC validation, then compare with cortical neuropil or cerebellar molecular layer staining before assigning NLGN2 localization (HPA antibodies; HPA tissue IHC; general IHC practice). |
| Colon or duodenum shows staining concentrated at enterocyte microvilli (HPA tissue IHC). | This is a reported medium IHC pattern in those cells (HPA tissue IHC); UniProt also reports NLGN2 protein detection in colon (UniProt Q8NFZ4 tissue specificity). | Record the microvillar compartment and intensity, and use a reported brain structure as an additional comparison when assessing assay performance (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing NLGN2 staining
A06595 · IHCImmunohistochemistry of NLGN2 in mouse brain tissue with NLGN2 antibody at 5 μg/ml.
A06595 · IFImmunofluorescence of NLGN2 in mouse brain tissue with NLGN2 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169992-NLGN2/tissue/cerebral+cortex#imid_19497423
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000169992-NLGN2/tissue/cerebellum#imid_19497418
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169992-NLGN2/tissue/colon#imid_19497333
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169992-NLGN2/tissue/liver#imid_19497342
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA055321; green = NLGN2, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA055321; green = NLGN2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NLGN2 IHC & IF
🧪Run cerebellum first and look for staining in molecular layer cells at the membrane and in the cytoplasm (HPA: Medium in cerebellar molecular layer cells). Use adipose tissue as the negative and score adipocytes, which HPA lists as not detected; nuclei within the positive cerebellar cells should remain unstained (HPA: Not detected in adipocytes; UniProt Q8NFZ4 subcellular localization).
Positive control tissue: Cerebellum (Molecular layer cells - cytoplasm/membrane, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NLGN2 in MCF-7, U-251MG, U2OS, NIH 3T3, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include no-primary or secondary-only and host-species-, isotype-, and clonality-matched controls, plus NLGN2 knockout tissue or peptide competition when an appropriate peptide is available (standard IHC practice). Check for endogenous peroxidase signal in cerebellum during chromogenic detection, and for tissue autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the A06595 mouse-brain IHC caption gives 5 µg/ml but leaves the fixative unreported (A06595 tissue-IHC caption). Antigen-retrieval dependence is also unreported, so retrieval conditions need empirical evaluation for paraffin sections (supplied evidence; standard IHC practice). The evidence does not establish whether frozen sections or IF are easier for NLGN2; check brain sections for background from endogenous peroxidase in IHC or autofluorescence in IF (supplied evidence; standard IHC/IF practice).
HPA tissue IHC evidence for NLGN2
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 |
|---|---|---|---|---|
| Cerebellum | Molecular layer cells - cytoplasm/membrane | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Colon | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Duodenum | Enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Testis | Sertoli cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NLGN2 IHC Tips
Troubleshoot chromogenic NLGN2 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting the signal.
How should I retrieve NLGN2 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then compare staining with an otherwise matched section processed without heat retrieval to judge whether retrieval improves signal (standard IHC practice). If staining remains weak, test a second retrieval condition on adjacent sections as a fallback, keeping antibody concentration and chromogen development constant (standard IHC practice). Score the expected membrane-associated neuropil pattern separately from diffuse cytoplasmic colour, because retrieval that increases total colour can also increase background (UniProt Q8NFZ4 localisation; standard IHC practice).
Could fixation explain variable NLGN2 staining across my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected mouse brain IHC caption reports 5 µg/ml antibody but does not state the fixative (A06595 tissue-IHC caption). Record the actual fixative, fixation interval, processing schedule and section age for every block before comparing staining intensity (standard IHC practice). Compare adjacent sections using the same citrate pH 6.0 retrieval and identical detection settings, changing only one processing variable at a time (page retrieval setting; standard IHC practice). Include a consistently processed reference block in each run so a weak result can be separated from day-to-day assay variation (standard IHC practice).
Which staining compartments should count as plausible NLGN2 signal?
Prioritise cell membrane and synaptic-region staining in brain, including neuropil, because NLGN2 is annotated at synaptic membranes and HPA reports medium cortical neuropil staining (UniProt Q8NFZ4 localisation; HPA tissue IHC). HPA also reports medium cytoplasm/membrane staining in cerebellar molecular-layer cells, so retain compartment labels when recording those cells (HPA tissue IHC). A separate HPA IF dataset assigns mitochondria as its approved subcellular location, which conflicts with the UniProt membrane annotation; treat mitochondrial-looking chromogenic granules as unresolved until independently validated (HPA subcellular; UniProt Q8NFZ4 localisation). Compare the pattern with a matched negative control and tissue architecture before calling puncta specific (standard IHC practice).
How do epitope position and processing affect NLGN2 IHC interpretation?
Check whether the antibody epitope lies in the extracellular region, residues 15–677, or cytoplasmic region, residues 699–835, across the transmembrane segment at 678–698 (UniProt Q8NFZ4 topology). Extracellular NLGN2 has annotated glycosylation sites at residues 98, 136 and 522, while the cytoplasmic region includes phosphoserines 713 and 718 (UniProt Q8NFZ4 processing). If the immunogen location is unavailable, do not attribute a staining change to glycosylation, phosphorylation or epitope masking without direct validation (standard IHC practice). UniProt lists 0 annotated isoforms here; that record alone does not establish what this antibody detects in processed tissue (UniProt Q8NFZ4 isoforms; standard IHC practice).
How can IF help assess whether my NLGN2 IHC pattern is specific?
On a separate IF preparation, multiplex NLGN2 with GPHN to assess association with inhibitory synaptic sites; UniProt reports their membrane colocalisation by similarity (UniProt Q8NFZ4 localisation). Choose fluorophores after checking the tissue’s autofluorescence in an unstained control, and place the weaker channel away from its strongest background (standard IF practice). Set permeabilisation according to epitope location: an extracellular epitope can be assessed without permeabilisation, whereas a cytoplasmic epitope requires access through the membrane (UniProt Q8NFZ4 topology; standard IF practice). Keep IF observations distinct from the paraffin IHC result, since HPA’s approved mitochondrial IF location conflicts with its tissue staining pattern and UniProt membrane annotation (HPA subcellular; HPA tissue IHC; UniProt Q8NFZ4 localisation).
How do I reduce diffuse brown signal without losing NLGN2 staining?
Run a no-primary control and inspect endogenous pigment and edge staining before adjusting the NLGN2 antibody (standard IHC practice). For peroxidase-based chromogenic detection, include an endogenous peroxidase block and keep DAB development consistent across matched sections; these are general workflow steps, not NLGN2-specific validation (standard IHC practice). Titrate the catalog antibody around the reported 5 µg/ml mouse-brain image condition only as an empirical comparison, since the caption does not establish an optimum for other samples (A06595 tissue-IHC caption; standard IHC practice). Judge improvement by preservation of membrane-associated signal and loss of diffuse colour in the same anatomical region (UniProt Q8NFZ4 localisation; standard IHC practice).
What should I score when NLGN2 stains both cells and neuropil? ⚠ ANSWER MARKED FOR VERIFICATION
Define anatomical regions and compartment rules before scoring, because cortical neuropil and cerebellar molecular-layer cells are both reported as positive patterns (HPA tissue IHC). For identifiable cells, record percentage positive and intensity on a 0–3 scale, then calculate an H-score from 0–300 (standard IHC practice). For neuropil, measure positive area or puncta density per mm² within the same region and subtract the matched negative-control signal (standard IHC practice). Normalise to analysed tissue area or the relevant cell count, and keep neuropil measurements separate from cell-based H-scores to avoid mixing compartments (standard IHC practice).
When is a brown NLGN2 signal credible rather than artefactual?
A credible brain pattern follows neuronal membranes or synaptic-rich neuropil and recurs in comparable sections, consistent with UniProt localisation and HPA cortical neuropil staining (UniProt Q8NFZ4 localisation; HPA tissue IHC; standard IHC practice). Examine the cellular context: HPA reports medium staining in cortical neuropil and cerebellar molecular-layer cells, but its reliability description notes only medium consistency with RNA data (HPA tissue IHC). Treat isolated nuclear colour, section-edge enhancement, necrotic deposits or signal retained in a no-primary control as reasons to investigate artefact (standard IHC practice). Check endogenous peroxidase control performance before assigning a DAB deposit to NLGN2, especially when its compartment disagrees with the expected membrane pattern (standard IHC practice; UniProt Q8NFZ4 localisation).
Boster reagents
Best NLGN2 / Neuroligin-2 IHC Antibodies
A06595 has mouse brain IHC and IF images (catalog image captions); IHC-P and IF are listed applications, with Human, Mouse and Rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-Neuroligin-2 NLGN2 Antibody
Cat # A06595
A06595 has an IHC image from mouse brain tissue at 5 μg/mL (catalog IHC image caption). Its IF image is also from mouse brain tissue, at 20 μg/mL (catalog IF image caption); Human, Mouse and Rat are listed as reactive species (catalog reactivity).
Which to pick: Choose A06595 for tissue IHC: IHC-P is listed, and its own IHC image shows mouse brain tissue at 5 μg/mL (catalog applications; A06595 IHC image caption). For fluorescence work, A06595 has an IF image from mouse brain at 20 μg/mL; ICC validation is unreported (A06595 IF image caption; catalog applications). A06595 lists Human, Mouse and Rat reactivity, though the supplied IHC image documents Mouse only; its fixative is unreported (catalog reactivity; A06595 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NFZ4 (NLGN2_HUMAN, Neuroligin-2).
- Human Protein Atlas. NLGN2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NLGN2 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. NLGN2 antibody validation summary (2 antibodies).
- A neuroligin-2-YAP axis regulates progression of pancreatic intraepithelial neoplasia. EMBO reports 2024 — PMC11014856.
- Clinical Relevance and Prognostic Value of the Neuronal Protein Neuroligin 2 in Breast Cancer. Frontiers in oncology 2021 — PMC8595097.
- Altered theta and beta oscillatory synchrony in a genetic mouse model of pathological anxiety. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021 — PMC12315943.
- Hippocampal neuroligin-2 overexpression leads to reduced aggression and inhibited novelty reactivity in rats. PloS one 2013 — PMC3579928.
- PubMed PMID:12669065 — UniProt-cited evidence.
- PubMed PMID:10718198 — UniProt-cited evidence.
- PubMed PMID:9278515 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NLGN2 (Neuroligin-2). Expected localisation Cortical neuropil; cerebellar cytoplasm/membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06595 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Molecular layer cells - cytoplasm/membrane); 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 A06595); 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-NLGN2, 5 μg/mL (datasheet A06595); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06595); Caveat: Staining–RNA consistency is medium (HPA tissue IHC); Isoform / epitope: No isoforms annotated; extracellular vs cytoplasmic epitope matters (UniProt); Regulation: Brain exceeds islet beta cells (UniProt). Evidence: UniProt Q8NFZ4, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A06595, PMC8595097, PMC3579928. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NLGN2/immunohistochemistry-ihc-guide.