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LYN / Tyrosine-protein kinase Lyn · IHC design guide
Design Immunohistochemistry for LYN
Plan chromogenic LYN IHC in paraffin sections using human tonsil as a positive reference (HPA tissue IHC; datasheet A01424-2). Compare staining by cell type and compartment, using the IHC-validated antibody at 0.5–1 μg/ml as a starting range (HPA tissue IHC; datasheet A01424-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LYN 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 | Membranous and cytoplasmic staining in selected tissues (HPA tissue IHC) | |
| Staining pattern | Germinal center cells show membranous and cytoplasmic signal (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01424-2) | |
| Positive control | Lymph node+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 A01424-2) | |
| Caveat | Staining varies by cell type within a tissue (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended LYN IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A01424-2) appears alongside published LYN protocols for colorectal cancer, melanoma, and cervical cancer tissue (PMC12174511; PMC6400685; PMC5342753).
| Sample | Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet A01424-2) |
| Fixation | Image fixative and duration unreported (datasheet A01424-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01424-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01424-2) |
| Primary antibody | Rabbit anti-LYN, 0.5-1μg/ml (datasheet A01424-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01424-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01424-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LYN-positive staining in germinal center cells of lymph node (HPA tissue IHC: High). HPA tissue profile: Distinct membranous and cytoplasmic expression in selected tissues at variable levels, most abundant in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A01424-2); the colorectal cancer protocol also specifies citrate at pH 6.0 (PMC12174511).
Section 2
What Is the Expected LYN Staining Pattern?
LYN is associated with the cell membrane and cytoplasm; UniProt also lists the Golgi apparatus and nucleus, and reports no transmembrane segment (UniProt P07948). In paraffin-section IHC, expect membranous and cytoplasmic staining chiefly in selected cells of lymphoid tissues (HPA: Enhanced reliability; most abundant in lymphoid tissues). Judge the pattern in identified cells rather than treating every stained cell in a section as equivalent (HPA: cell-specific tissue IHC).
| Membranous and cytoplasmic staining in lymph-node or tonsil germinal center cells. | This matches the reported compartment pattern and High staining in those cells (HPA: tissue IHC). Compare with local background before scoring intensity; the HPA level describes those cells, not every cell in the section. |
| Predominantly nuclear staining, with little convincing membrane or cytoplasmic signal. | Investigate a compartment mismatch or artefact against the HPA tissue-IHC pattern (HPA: membranous and cytoplasmic). Nuclear LYN remains biologically possible: UniProt reports nuclear localisation and increased accumulation when CRM1-mediated export is inhibited (UniProt P07948). Nuclear staining alone is therefore not proof of specificity. |
| Strong staining in a cell population listed as undetected, such as adipocytes in adipose tissue. | Consider cross-reactivity, endogenous detection activity, or misidentified cells before assigning LYN positivity (HPA: adipocytes, Not detected). Score the named cell population specifically; staining in a different cell within the same tissue does not overturn that HPA observation. |
| Diffuse chromogen across tissue, extracellular spaces, or the whole section without cell boundaries. | A broad haze cannot establish LYN localisation against the distinct membranous and cytoplasmic reference pattern (HPA: tissue IHC). General IHC practice: compare a no-primary control and examine blocking, washing, and detection chemistry before calling weak cells positive. |
| No signal in germinal center cells of a lymph node or tonsil control. | Treat the run as unresolved before scoring test sections: both cell populations are reported High (HPA: lymph node and tonsil). General IHC practice: check tissue preservation, retrieval, antibody incubation, and the detection system; absence alone does not establish biological loss of LYN. |
💡Expected LYN appearanceCall a convincing positive result when identified lymph-node or tonsil germinal center cells show distinct membranous and cytoplasmic chromogen at a clearly visible level (HPA: High); uniform haze or strong signal confined to HPA-undetected adipocytes is suspect (HPA: tissue IHC).
| Cell and tissue choice (HPA: tissue IHC). | Lymph-node and tonsil germinal center cells, placental Hofbauer cells, and splenic red-pulp cells are High; marrow hematopoietic cells are Medium (HPA: tissue IHC). These are cell-specific reference observations, not whole-section intensity standards. |
| Compartment and topology (UniProt P07948; HPA: tissue IHC). | LYN has no transmembrane segment (UniProt P07948), while tissue IHC shows membranous and cytoplasmic expression (HPA: tissue IHC). A membrane outline is compatible with LYN; do not require a transmembrane-spanning appearance. |
| Antibody validation (HPA: antibody records). | IHC staining is marked Enhanced for HPA001231 and CAB004492 (HPA: antibody records). The tissue profile also has Enhanced reliability based on staining and RNA consistency (HPA: tissue IHC); neither designation validates every apparent positive cell. |
| Q: What should IF/ICC show? (HPA: subcellular ICC-IF). | A: Mainly plasma-membrane signal, with additional Golgi and vesicle signal (HPA: subcellular ICC-IF). This is a localisation reference for IF/ICC, not an IHC-P staining intensity standard or a protocol option. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive control is blank (HPA: lymph-node or tonsil germinal center cells, High). | Possible failed assay step or unsuitable control section; the slide alone cannot distinguish them (general IHC practice). | Verify control morphology and the documented IHC-P workflow, then check retrieval, primary incubation, and detection with appropriate run controls (general IHC practice). Do not score patient or experimental sections until the control works. |
| Chromogen covers the slide as a diffuse haze. | Incomplete blocking or washing, or detection background, can obscure cell boundaries (general IHC practice). | Compare a no-primary control; review blocking, washes, and detection development, then reassess whether staining follows the HPA membranous and cytoplasmic pattern (HPA: tissue IHC; general IHC practice). |
| Adipocytes stain strongly in adipose tissue (HPA: Not detected in adipocytes). | Possible cross-reactivity, endogenous detection activity, or cell misidentification (general IHC practice). | Confirm cell identity and compare no-primary and reagent controls; score adipocytes separately from other cells in the section (general IHC practice; HPA: adipocyte-specific observation). |
| Only nuclei stain in otherwise expected positive cells. | This differs from the usual HPA tissue pattern, though regulated nuclear LYN is reported (HPA: tissue IHC; UniProt P07948). | Review controls and cell morphology, then compare membrane and cytoplasmic signal with the HPA reference; report nuclear-only staining as uncertain unless independently supported. |
| Weak or absent signal is judged in an HPA low-staining population. | Low baseline staining can limit the usefulness of that population as a control (HPA: low in cerebral-cortex endothelial cells and several other listed populations). | Include a named High cell population, such as tonsil germinal center cells, for run interpretation (HPA: tissue IHC). Avoid calling assay failure from a low-staining population alone. |
| Signal appears in a tissue that also contains HPA-undetected cells. | Mixed cell populations can make a whole-section positive call misleading (HPA: cell-specific tissue IHC). | Record the stained cell type and compartment. Compare each population with its HPA entry; for example, kidney glomerular cells are listed as Not detected (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing LYN staining
A01424-2 · IHCIHC analysis of Lyn using anti-Lyn antibody (A01424-2). Lyn was detected in paraffin-embedded section of mouse intestine tissue . Heat media
A01424-2 · IHCIHC analysis of Lyn using anti-Lyn antibody (A01424-2). Lyn was detected in paraffin-embedded section of rat spleen tissue . Heat mediated a
A01424Y508 · IHCImmunohistochemistry analysis of paraffin-embedded human brain, using Lyn (Phospho-Tyr507) Antibody. The picture on the right is blocked wit
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000254087-LYN/tissue/cerebral+cortex#imid_517082
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000254087-LYN/tissue/colon#imid_517016
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000254087-LYN/tissue/liver#imid_1458035
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000254087-LYN/tissue/kidney#imid_517018
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001231; green = LYN, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001231; green = LYN, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LYN IHC & IF
🧪Run lymph node first: germinal center cells should stain (HPA: High in lymph node germinal center cells). Use adipose tissue as the negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the lymph node slide, cells without specific staining should show only counterstain and background, but do not presume every cell outside germinal centers is LYN-negative.
Positive control tissue: Lymph node (Germinal center cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LYN in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, host-matched nonimmune rabbit IgG matched to the primary antibody’s clonality, and LYN-knockout material as a biological negative (selected-SKU caption: rabbit anti-Lyn primary). Block endogenous peroxidase and check for endogenous biotin signal when using the caption’s biotin–streptavidin/DAB detection in lymph node (selected-SKU caption: biotinylated secondary, streptavidin–biotin complex and DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The caption reports citrate retrieval at pH 6 for 20 minutes in mouse intestine; it does not establish whether retrieval is required in lymph node (selected-SKU caption: mouse intestine IHC). No supplied matched evidence establishes that frozen sections or IF are easier; for lymph node IHC, assess endogenous peroxidase and biotin background with the stated detection system (selected-SKU caption: biotin–streptavidin/DAB).
HPA tissue IHC evidence for LYN
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | Hofbauer cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LYN IHC Tips
Troubleshoot LYN staining in paraffin sections by checking retrieval, cell identity and subcellular pattern before comparing chromogenic signal across samples.
How should I adjust retrieval when LYN staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 (datasheet A01424-2). The selected paraffin-section example used 20 minutes of retrieval, followed by 1 µg/mL primary antibody overnight at 4°C (caption A01424-2). Compare the weak section with lymph-node germinal-center cells processed in the same run, since those cells show high LYN staining (HPA: high in lymph-node germinal-center cells). If both are weak, adjust heating duration in small steps while holding antibody concentration constant (standard IHC practice). Accept the setting that preserves morphology and distinct membrane or cytoplasmic signal without increasing negative-control staining (standard IHC practice; HPA: membranous and cytoplasmic profile).
Could fixation explain inconsistent LYN staining between paraffin blocks?
The selected paraffin-section caption does not state its fixative, so LYN-specific fixation sensitivity is unknown (caption A01424-2). Record each block’s fixative and fixation duration, and compare sections cut, retrieved and stained in the same run (standard IHC practice). Use the documented citrate retrieval at pH 6 for 20 minutes as the initial common condition (caption A01424-2). Include a lymph-node germinal-center control to check whether the run detects a cell population with high LYN staining (HPA: high in lymph-node germinal-center cells). If blocks differ, assess morphology and control staining before attributing the difference to LYN expression (standard IHC practice).
Should LYN appear at the membrane, in cytoplasm, or in nuclei?
Score membrane-associated and cytoplasmic staining separately: both occur in tissue IHC, with particularly abundant signal in lymphoid tissues (HPA: tissue IHC profile). LYN has no transmembrane segment, so a membrane pattern does not require staining that spans the plasma membrane (UniProt P07948 topology). Golgi and vesicular localisation also have support from subcellular imaging, making a limited perinuclear pattern plausible (HPA: subcellular profile). Nuclear localisation is reported, including increased accumulation when CRM1-mediated export is inhibited (UniProt P07948 subcellular localisation). Verify any dominant nuclear pattern against controls and cell identity before assigning it to LYN (standard IHC practice).
Can this antibody distinguish LYN isoforms or modified epitopes in IHC?
LYN has 2 listed isoforms, while the supplied product caption does not identify the antibody’s epitope (UniProt P07948 isoforms; caption A01424-2). Therefore, the staining alone cannot establish isoform specificity; obtain the immunogen sequence and map it against both isoforms before making that claim (standard IHC practice). LYN contains SH3 residues 63–123, SH2 residues 129–226 and kinase residues 247–501 (UniProt P07948 domains). Its listed phosphorylation sites include Ser-11, Tyr-193 and Tyr-306 (UniProt P07948 modified residues). Treat altered staining after pathway perturbation as an epitope-accessibility question until orthogonal evidence separates protein abundance from modification state (standard IHC practice).
How can I check a LYN pattern by multiplex immunofluorescence?
For a separate IF assay, pair LYN with a marker of the expected cell population, such as CD68 for Hofbauer macrophages in placenta (HPA: high in placental Hofbauer cells; standard IF practice). Choose a far-red fluorophore after imaging an unstained section to identify tissue autofluorescence in each channel (standard IF practice). LYN has no transmembrane segment and is reported at the plasma membrane and in cytoplasmic compartments, so compare staining with and without mild permeabilisation to reveal epitopes facing the cytoplasm (UniProt P07948 topology; HPA: subcellular profile; standard IF practice). Acquire single-label controls before interpreting overlap between channels (standard IF practice). Use IF localisation as a cross-check of the chromogenic IHC pattern, with its own controls and optimisation (standard IF practice).
What should I check when LYN DAB staining looks diffuse?
Check a no-primary section for endogenous enzyme signal and nonspecific chromogen deposition before changing the LYN antibody (standard IHC practice). Apply a peroxidase block appropriate to the tissue, since the selected method uses DAB chromogen (standard IHC practice; caption A01424-2). The selected method also uses 10% goat serum and a streptavidin–biotin detection complex, so check blocking and endogenous biotin if background follows tissue structures (caption A01424-2; standard IHC practice). Titrate primary antibody around the documented 1 µg/mL condition while keeping retrieval fixed (caption A01424-2; standard IHC practice). Judge improvement by distinct cellular staining and a clean no-primary control (standard IHC practice).
How should I quantify LYN staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC shows variable membranous and cytoplasmic LYN staining (HPA: tissue IHC profile). For comparable cells, record the percentage positive and intensity, or calculate an H-score on the 0–300 scale (standard IHC practice). For infiltrating populations, report positive-cell density per mm² of viable tissue and identify the denominator used (standard IHC practice). Keep threshold, chromogen development and image acquisition settings consistent across the comparison (standard IHC practice). Normalise scores to the relevant cell population or viable tissue area, since high staining in lymphoid cells can change a whole-section average (HPA: high in lymphoid tissues; standard IHC practice).
How do I separate genuine LYN staining from artefact?
Expect interpretable staining mainly along membranes and in cytoplasm, and check which cells carry the signal (HPA: tissue IHC profile). Germinal-center cells in lymph node provide a high-staining comparator, while adipocytes in adipose tissue are reported as not detected (HPA: lymph-node germinal-center cells high; HPA: adipose adipocytes not detected). A nuclear signal warrants closer review because nuclear LYN is reported, but its accumulation depends on cellular context (UniProt P07948 subcellular localisation). Exclude section edges and necrotic areas from interpretation, and inspect a no-primary control for endogenous enzyme or DAB artefact (standard IHC practice). Report the compartment and cell population rather than treating any brown deposit as LYN-positive (standard IHC practice).
Boster reagents
Best LYN / Tyrosine-protein kinase Lyn IHC Antibodies
Catalog antibodies cover IHC and IF/ICC in human, mouse and rat (catalog: applications and reactivity). IHC images show paraffin sections of intestine, spleen, tonsil and brain (catalog: IHC image captions).
Real IHC data
Anti-Lyn Antibody ®
Cat # A01424-2
Real IHC data
Anti-Phospho-Lyn (Y508) Antibody
Cat # A01424Y508
A01424-2 has IHC images from paraffin sections of mouse intestine and spleen, rat spleen and human tonsil (catalog: A01424-2 image captions). A01424Y508 has a phospho-Lyn IHC image from paraffin-embedded human brain with phosphopeptide blocking (catalog: A01424Y508 image caption).
Which to pick: For tissue IHC across human, mouse and rat, choose A01424-2: its own paraffin-section captions report citrate retrieval, and the fixative is unreported (catalog: A01424-2 applications, reactivity and image captions). For IF/ICC, P01424-1 is a rabbit monoclonal listed for both applications in human and mouse (catalog: P01424-1 clone, applications and reactivity). For phospho-Lyn IF across human, mouse and rat, A01424Y508 lists IF; its own IHC caption supports paraffin-embedded human brain, with fixative unreported (catalog: A01424Y508 applications, reactivity and image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07948 (LYN_HUMAN, Tyrosine-protein kinase Lyn).
- Human Protein Atlas. LYN tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LYN subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the Golgi apparatus and vesicles..
- Human Protein Atlas. LYN antibody validation summary (2 antibodies).
- Role of Lyn Immunohistochemical Staining in Progression of Colorectal Carcinoma. Asian Pacific journal of cancer prevention : APJCP 2025 — PMC12174511.
- Lyn Kinase Promotes the Proliferation of Malignant Melanoma Cells through Inhibition of Apoptosis and Autophagy via the PI3K/Akt Signaling Pathway. Journal of Cancer 2019 — PMC6400685.
- Tyrosine kinase LYN is an oncotarget in human cervical cancer: A quantitative proteomic based study. Oncotarget 2016 — PMC5342753.
- Minocycline binds and inhibits LYN activity to prevent STAT3-meditated metastasis of colorectal cancer. International journal of biological sciences 2022 — PMC8990469.
- PubMed PMID:3561390 — UniProt-cited evidence.
- PubMed PMID:8125304 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LYN (Tyrosine-protein kinase Lyn). Expected localisation Membranous and cytoplasmic staining in selected tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01424-2). Validated IHC antibody: A01424-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lymph node (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet A01424-2); 2. Fixation: Image fixative and duration unreported (datasheet A01424-2); 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: Citrate pH 6 , 20 min (datasheet A01424-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01424-2); 8. Primary antibody: Rabbit anti-LYN, 0.5-1μg/ml (datasheet A01424-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01424-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01424-2); Caveat: Staining varies by cell type within a tissue (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression regulation is not specified (UniProt). Evidence: UniProt P07948, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A01424-2, PMC12174511, PMC6400685. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LYN/immunohistochemistry-ihc-guide.