LYZ / Lysozyme C · IHC design guide

Design Immunohistochemistry for LYZ

Plan chromogenic LYZ IHC in paraffin sections using Paneth cells and lung macrophages as high-staining references (HPA tissue IHC). Score cytoplasmic staining and interpret its distribution with LYZ secretion in mind (HPA tissue IHC; UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for LYZ (IHC for LYZ): expected localisation Cytoplasmic tissue staining; LYZ is secreted (HPA tissue IHC; UniProt), antibody PB9663, validated IHC image, and IHC protocol steps
Printable LYZ IHC protocol sheet — expected localisation Cytoplasmic tissue staining; LYZ is secreted (HPA tissue IHC; UniProt), antibody PB9663, controls and protocol steps. Open the full LYZ IHC guide →

LYZ 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 tissue staining; LYZ is secreted (HPA tissue IHC; UniProt)
Staining pattern High cytoplasmic staining in marrow cells, Paneth cells and lung macrophages (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet PB9663)
Positive control ⓘ Bone marrow+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 PB9663)
Caveat Secreted LYZ may appear away from expressing cells (HPA tissue IHC; UniProt)
Regulation No expression regulator specified (UniProt)
Isoform / epitope No isoforms annotated; mature chain spans residues 19–148 (UniProt)
Section 1

Recommended LYZ IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published LYZ staining methods for porcine intestine, mouse kidney, and human intestinal biopsies (PMC13317171; PMC11101254; PMC2776509).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human intestinal cancer tissue; fixative not specified (datasheet PB9663)
FixationImage fixative and duration unreported (datasheet PB9663); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet PB9663); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet PB9663)
Primary antibodyRabbit anti-LYZ, 0.5-1μg/ml (datasheet PB9663)
Primary incubationOvernight at 4 °C (datasheet PB9663)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9663)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultLYZ-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: High cytoplasmic expression in selected cells types including cells in bone marrow, Paneth cells and a subset of cells in salivary gland. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: PB9663); the mouse kidney study used microwave retrieval in water (PMC11101254).
Section 2

What Is the Expected LYZ Staining Pattern?

In paraffin section IHC, LYZ should stain the cytoplasm of selected hematopoietic cells, Paneth cells, lung macrophages, and some glandular cells (HPA: tissue IHC). LYZ is secreted and has no transmembrane segment (UniProt P61626: subcellular location and topology). HPA rates the tissue staining evidence Enhanced, while reporting medium consistency between staining and RNA data because secreted protein can appear away from its site of synthesis (HPA: tissue IHC reliability).

What am I looking at on my slide?
Strong cytoplasmic staining in duodenal or small intestinal Paneth cells, lung macrophages, or bone marrow hematopoietic cells (HPA: High in these cells).This matches the reported positive pattern. Judge the named cell population and its cytoplasm, rather than treating uniform staining across every cell in the section as the expected result (HPA: tissue IHC profile).
Predominantly nuclear staining, with little cytoplasmic signal, in a tissue chosen for positive IHC (HPA: cytoplasmic tissue IHC profile).This does not match the reported tissue IHC pattern; review morphology, controls, and detection background before calling the cells LYZ positive (HPA: tissue IHC profile). The separate ICC-IF record lists nucleoplasm only as an uncertain additional location (HPA: subcellular ICC-IF).
Strong staining in adipocytes or another cell type reported as not detected (HPA: adipose tissue and breast adipocytes; tissue IHC).Treat an unexpected cell pattern as provisional: cross-reactivity or endogenous detection activity is possible (general IHC practice). Confirm cell identity and compare a no-primary control; secretion means protein location need not mirror cellular RNA expression (HPA: reliability description; UniProt P61626: Secreted).
Diffuse color across tissue, including cells expected to have little or no staining (HPA: low and not-detected tissue IHC entries).The contrast needed to recognize selected positive cells has been lost (HPA: tissue IHC profile). General IHC practice: check no-primary background, blocking, washes, detection conditions, and counterstain before interpreting faint cell-specific signal.
No signal in correctly identified Paneth cells, lung macrophages, or bone marrow hematopoietic cells (HPA: High in these populations).An expected positive population is missing, so a negative result elsewhere is difficult to interpret (HPA: tissue IHC). General IHC practice: verify tissue identity and controls, then review antibody dilution, retrieval, detection, and reagent performance.
💡Expected LYZ appearanceCall an IHC result consistent with LYZ when selected Paneth cells, lung macrophages, or bone marrow hematopoietic cells show strong cytoplasmic staining (HPA: High; cytoplasmic profile); widespread nuclear color or uniform staining of unrelated cells warrants background and specificity checks (HPA: tissue IHC profile; general IHC practice).
How each factor affects the staining
Cell population and tissue (HPA: tissue IHC)High staining is reported in bone marrow hematopoietic cells, duodenal and small intestinal Paneth cells, and lung macrophages; appendix, kidney, salivary gland, and stomach have specified medium-staining populations (HPA: tissue IHC). Choose and score the relevant cells, not the whole section.
Secretion and interpretation (UniProt P61626: Secreted; HPA: reliability description)LYZ lacks a transmembrane segment and is secreted (UniProt P61626: topology and location). HPA cautions that protein and RNA tissue locations can differ for this reason, and describes medium staining–RNA consistency despite Enhanced tissue IHC reliability (HPA: tissue IHC).
Processing and epitope information (UniProt P61626: sequence features)The precursor is 148 amino acids, with a signal sequence at residues 1–18 and a mature chain at 19–148 (UniProt P61626: processing). The supplied record does not locate this antibody’s epitope; these sequence features alone cannot predict its retrieval conditions.
Antibody evidence (HPA: antibody validation and tissue IHC reliability)HPA lists Enhanced IHC status for HPA048284 and CAB000055 (HPA: antibody validation). That supports the reported tissue pattern, while the medium RNA–staining consistency remains a limitation when interpreting individual unexpected cells (HPA: reliability description).
IF/ICC Q&A: where can LYZ appear? (HPA: subcellular ICC-IF)HPA lists Golgi apparatus as supported, actin filaments as uncertain, and nucleoplasm as an uncertain additional ICC-IF location (HPA: subcellular ICC-IF). These cell imaging observations should be interpreted separately from the cytoplasmic tissue IHC pattern (HPA: tissue IHC profile).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
A known positive population is blank (HPA: High in Paneth cells, lung macrophages, and bone marrow hematopoietic cells).General IHC practice: an inactive detection step, unsuitable working conditions, or an incorrectly identified cell population could explain the result; the supplied sources do not establish LYZ-specific fixation sensitivity.General IHC practice: confirm morphology and positive control performance; then review reagent function, antibody dilution, and retrieval settings against the antibody’s IHC instructions.
Only weak signal appears in a medium or low HPA population (HPA: tissue IHC levels).The selected population has less reported staining than the High reference populations (HPA: tissue IHC). A weak section alone cannot distinguish low target abundance from a workflow problem.Include a High reference population from the HPA tissue IHC record and compare matched sections under the same general IHC detection conditions (HPA: High populations; general IHC practice).
Unexpected widespread nuclear staining appears in tissue IHC (HPA: cytoplasmic tissue IHC profile).The pattern conflicts with HPA tissue IHC; background or nonspecific binding is possible (HPA: tissue IHC profile; general IHC practice). HPA’s uncertain nucleoplasmic ICC-IF entry does not validate widespread nuclear tissue staining (HPA: subcellular ICC-IF).General IHC practice: compare no-primary and positive controls, inspect counterstain and cell boundaries, and reassess before assigning nuclear LYZ expression.
Color develops in a no-primary control (general IHC practice).Signal without the primary antibody points to detection background, which can include endogenous enzyme activity in chromogenic IHC (general IHC practice).General IHC practice: review the chromogen system, endogenous activity blocking where applicable, secondary detection, and wash conditions before interpreting the test section.
Adipocytes stain strongly despite their not-detected HPA entries (HPA: adipose tissue and breast adipocytes).The observed cell pattern differs from those HPA tissue IHC entries; cross-reactivity, detection background, or mistaken cell identification is possible (HPA: tissue IHC; general IHC practice).General IHC practice: verify morphology, compare a no-primary control, and seek agreement with an independent antibody or another appropriate specificity control.
Staining and RNA results seem to disagree across tissues (HPA: medium staining–RNA consistency).LYZ is secreted (UniProt P61626: subcellular location); HPA says the tissue locations of its RNA and protein may differ (HPA: reliability description).Interpret the stained cell population against HPA’s tissue IHC observations and the section’s controls; avoid using RNA abundance alone to declare IHC staining correct or erroneous (HPA: tissue IHC reliability; general IHC practice).

Sample controls for LYZ IHC & IF

🧪Run duodenum first: Paneth cells should stain strongly (HPA: High in duodenal Paneth cells). Use adipose tissue as the negative tissue, scoring adipocytes rather than any resident immune cells (HPA: Not detected in adipocytes); on the duodenum slide, adjacent non-Paneth epithelium should lack the strong Paneth-cell pattern (HPA: High in Paneth cells).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LYZ in CACO-2, Hep-G2, with annotated localisation: Golgi apparatus (supported), Actin filaments (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a rabbit IgG control matched to the primary antibody’s format and clonality; and LYZ-knockout material or peptide preadsorption if a suitable peptide is available (selected PB9663 caption: rabbit primary; standard IHC practice). For chromogenic duodenal sections, quench endogenous peroxidase and check for endogenous biotin if using the caption’s biotin-based detection system (selected PB9663 caption: biotinylated secondary and SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9663 paraffin-section caption does not state its fixative (selected PB9663 caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for every LYZ antibody or specimen (selected PB9663 caption). There is insufficient matched evidence to say frozen sections or IF are easier; in duodenum, secreted LYZ may produce extracellular signal that complicates cell-localization scoring (UniProt P61626: secreted; HPA: High in duodenal Paneth cells).

HPA tissue IHC evidence for LYZ

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Secreted protein, tissue location of RNA and protein is expected to differ.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Bone marrow Hematopoietic cells High Protein (IHC) HPA →
Duodenum Paneth cells High Protein (IHC) HPA →
Lung Macrophages High Protein (IHC) HPA →
Small intestine Paneth cells High Protein (IHC) HPA →
Appendix Non-germinal center cells Medium Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Adrenal gland Glandular cells Not detected Protein (IHC) HPA →
Breast Adipocytes Not detected Protein (IHC) HPA →
Caudate Glial cells Not detected Protein (IHC) HPA →
Cerebellum Cells in granular layer Not detected Protein (IHC) HPA →
Section 3

Advanced LYZ IHC Tips

Troubleshoot LYZ staining in paraffin sections by checking retrieval, tissue handling, cell identity, and the distribution of this secreted protein (UniProt P61626; HPA tissue IHC).

What retrieval should I use when LYZ staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin-section IHC (datasheet PB9663). The selected image used that retrieval, followed by 1 μg/ml primary antibody overnight at 4°C (caption PB9663). If staining is weak, compare a longer heating interval against the initial run while keeping the antibody concentration and detection steps fixed (standard IHC practice). Include a known positive section containing Paneth cells or macrophages and inspect morphology before accepting stronger staining as improved retrieval (HPA tissue IHC; standard IHC practice). Record the heating time and cooling conditions so the comparison can be repeated (standard IHC practice).
How should I troubleshoot variable LYZ staining after fixation?
The selected paraffin-section caption does not report its fixative, so target-specific fixation sensitivity is unknown (caption PB9663). Compare sections with documented fixation histories using the same EDTA pH 8.0 retrieval and 1 μg/ml primary antibody conditions before changing several variables together (datasheet PB9663; caption PB9663; standard IHC practice). Keep section thickness, retrieval heating, and chromogen development consistent, and check whether weak areas also show poor tissue preservation (standard IHC practice). Because LYZ is secreted, interpret staining in its tissue context, but do not use that property to infer a particular fixation effect (UniProt P61626; standard IHC practice).
Where should convincing LYZ staining appear in a paraffin section?
Expect staining in selected cells rather than uniform positivity: HPA reports high cytoplasmic expression in Paneth cells and high staining in lung macrophages and bone-marrow hematopoietic cells (HPA tissue IHC). LYZ is secreted and has no transmembrane segment, so cellular staining can coexist with extracellular signal (UniProt P61626 topology; standard IHC interpretation). Map the stained cells against morphology and a counterstain before assigning the signal to epithelium, inflammatory cells, or extracellular material (standard IHC practice). If signal is confined to nuclei or spreads evenly across unrelated cell populations, repeat the assessment with a positive tissue and a no-primary control (HPA tissue IHC; standard IHC practice).
Could an isoform or epitope difference explain a negative LYZ stain?
The supplied record lists 0 isoforms, a signal peptide at residues 1–18, and a mature chain at residues 19–148 (UniProt P61626 processing; UniProt P61626 isoforms). It also lists 0 glycosylation sites and 0 modified residues, but the antibody epitope is not supplied (UniProt P61626 modifications; caption PB9663). Therefore, do not attribute a negative section to an unreported isoform or a specific modified epitope (UniProt P61626 isoforms; standard IHC interpretation). First check retrieval, positive-control staining, antibody incubation, and detection; if those work, request epitope information before drawing an epitope-specific conclusion (datasheet PB9663; standard IHC practice).
How can I assess LYZ by IF alongside the chromogenic IHC result?
Treat IF as a separate assay and pair LYZ with a marker identifying the expected cell type, such as a macrophage marker when examining macrophage-rich fields (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel if the specimen has strong short-wavelength autofluorescence (standard IF practice). For an intracellular epitope in the secretory pathway, test gentle permeabilisation and compare it with an unpermeabilised condition, since the antibody epitope is unspecified and mature LYZ is secreted (UniProt P61626 processing; standard IF practice). Use single-label and secondary-only controls to assess bleed-through and nonspecific fluorescence; do not transfer the paraffin-section result to IF without validation (caption PB9663; standard IF practice).
What should I check when DAB background obscures LYZ-positive cells?
The selected IHC example used a 10% goat-serum block, a biotinylated secondary antibody, a streptavidin–biotin detection complex, and DAB (caption PB9663). If diffuse brown signal obscures cells, inspect a no-primary control, verify the peroxidase block, and shorten DAB development in a controlled comparison (standard chromogenic IHC practice). Check for endogenous biotin background when using the selected detection chemistry, especially if the no-primary control stains (caption PB9663; standard IHC practice). Review section edges, folds, and damaged areas separately from intact tissue, and judge improvements against expected Paneth-cell or macrophage staining (HPA tissue IHC; standard IHC practice).
How should I quantify LYZ staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring, because HPA reports high staining in Paneth cells and lung macrophages, while LYZ is secreted (HPA tissue IHC; UniProt P61626). For cell-associated DAB signal, report the percentage of positive target cells or an H-score based on intensity and percentage; use positive-cell density per mm² when tissue area is the appropriate denominator (standard IHC quantification practice). Normalise each comparison to the same annotated tissue area or the same eligible cell population, and exclude folds and necrotic regions by a preset rule (standard IHC quantification practice). Keep retrieval, counterstain, imaging, and scoring thresholds consistent across sections (standard IHC practice).
How can I distinguish genuine LYZ positivity from staining artefacts?
A convincing result fits cell morphology and expected distribution, including Paneth cells or macrophages, rather than appearing indiscriminately across a section (HPA tissue IHC; standard IHC interpretation). LYZ is secreted, so extracellular material may contain signal; assess it alongside nearby cells instead of treating every deposit as a positive cell (UniProt P61626; standard IHC interpretation). Be cautious with isolated nuclear staining because the tissue IHC profile emphasizes cytoplasmic expression; HPA subcellular data describe nucleoplasm staining as uncertain (HPA tissue IHC; HPA subcellular). Compare intact tissue with edges and necrotic areas, and check no-primary staining for endogenous peroxidase or detection-related artefact before scoring (standard chromogenic IHC practice).
Boster reagents

Best LYZ / Lysozyme C IHC Antibodies

Anti-LYZ antibodies have IHC images from paraffin sections of human intestinal cancer and mouse kidney (catalog IHC captions), plus IF images from human and mouse samples (catalog IF captions).

Real IHC data IHC analysis of Lysozyme using anti-Lysozyme antibody (PB9663). Lysozyme was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Lysozyme Antibody (PB9663) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-Lysozyme/LYZ Antibody ®
Cat # PB9663
Real IHC data Immunohistochemical analysis of paraffin-embedded mouse kidney, using Lysozyme Antibody.
Anti-Lysozyme LYZ Rabbit Monoclonal Antibody
Cat # M01811

PB9663 has an IHC image from paraffin-embedded human intestinal cancer tissue and IF images from human ileum, human colon organoids, mouse ileum, and mouse ileum organoids (PB9663 image captions). M01811 has an IHC image from paraffin-embedded mouse kidney and an IF image from mouse colon; its application list also includes ICC (M01811 image captions; catalog applications).

Which to pick: For human tissue IHC, PB9663 has a paraffin-section image using 1 μg/ml antibody; the caption does not report the fixative (PB9663 IHC caption). For IF/ICC, M01811 is the rabbit monoclonal option with both applications listed, while PB9663 has IF images from human and mouse samples (M01811 catalog applications and clone; PB9663 IF captions). For work across human, mouse, and rat, both list those species as reactive, but their own IHC figure captions document only human intestinal cancer for PB9663 and mouse kidney for M01811; neither caption reports the fixative (catalog reactivity; PB9663 and M01811 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P61626 (LYSC_HUMAN, Lysozyme C).
  2. Human Protein Atlas. LYZ tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. LYZ subcellular location (ICC-IF): Mainly localized to the Golgi apparatus and actin filaments. In addition localized to the nucleoplasm..
  4. Human Protein Atlas. LYZ antibody validation summary (3 antibodies).
  5. Dynamic distribution of Paneth cells in pig intestine under normal and infection conditions. BMC veterinary research 2026 — PMC13317171.
  6. Lysozyme promotes renal fibrosis through the JAK/STAT3 signal pathway in diabetic nephropathy. Archives of medical science : AMS 2024 — PMC10895955.
  7. NET-Related Gene as Potential Diagnostic Biomarkers for Diabetic Tubulointerstitial Injury. Journal of diabetes research 2024 — PMC11101254.
  8. Mucosal gene expression of antimicrobial peptides in inflammatory bowel disease before and after first infliximab treatment. PloS one 2009 — PMC2776509.
  9. PubMed PMID:2971592 — UniProt-cited evidence.
  10. PubMed PMID:3413092 — UniProt-cited evidence.
  11. PubMed PMID:2829884 — UniProt-cited evidence.