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LCP1 / Plastin-2 · IHC design guide
Design Immunohistochemistry for LCP1
Plan paraffin-section LCP1 IHC around cytoplasmic immune-cell staining, with tonsil germinal-center cells as a positive reference (HPA tissue IHC). This guide covers fixation consistency, antibody titration, chromogenic detection, and cell-specific scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LCP1 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 | Cytoplasm in immune cells (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic staining in immune cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A03361) | |
| Positive control | Appendix+4 more · see all | |
| 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); validate conditions locally. | |
| Caveat | Strong lung macrophage staining can complicate cell scoring (HPA tissue IHC) | |
| Regulation | TCR costimulation shifts LCP1 to synapses (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified; cytoplasmic target (UniProt) |
Section 1
Recommended LCP1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A03361). These published methods describe LCP1 staining in breast, sarcoma, and urological cancer samples (PMC11229261; PMC11974013; PMC13266032).
| Sample | Paraffin-embedded human B lymphocytic tumor tissue; fixative not specified (datasheet A03361) |
| Fixation | Image fixative and duration unreported (datasheet A03361); 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 A03361) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03361) |
| Primary antibody | Rabbit anti-LCP1, 0.5-1μg/ml (datasheet A03361) |
| Primary incubation | Overnight at 4 °C (datasheet A03361) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03361) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LCP1-positive staining in germinal center cells of appendix (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A03361); the three published excerpts do not state retrieval conditions (PMC11229261; PMC11974013; PMC13266032).
Section 2
What Is the Expected LCP1 Staining Pattern?
LCP1 is a cytoskeletal protein without a transmembrane segment (UniProt P13796 topology). In paraffin section IHC, expect selective cytoplasmic staining in immune cells; HPA rates the tissue pattern Enhanced for agreement between staining and RNA data (HPA: tissue IHC). UniProt also places LCP1 at cell projections and junctions, while HPA ICC-IF supports a plasma membrane location; assess those findings in their respective applications (UniProt P13796 subcellular; HPA: ICC-IF).
| Strong cytoplasmic staining in lung macrophages, bone marrow hematopoietic cells, or lymphoid germinal center cells (HPA: High in each). | This matches the selective immune cell pattern reported for tissue IHC (HPA: tissue IHC). Judge the stained cells and their compartment together: a positive area alone does not establish that the expected cells are positive (general IHC interpretation). |
| Predominantly nuclear staining, with little staining in the expected immune cell cytoplasm (HPA: tissue IHC pattern). | A nuclear dominant result conflicts with HPA tissue IHC and UniProt cytoskeletal localization (HPA: tissue IHC; UniProt P13796 subcellular). Treat it as suspect and check controls, antibody dilution, and detection conditions before assigning LCP1 localization (general IHC practice). |
| Staining concentrated in adipocytes or bronchial respiratory epithelial cells (HPA: Not detected in those cells). | These cells are reported as negative in the sampled HPA tissues (HPA: adipose tissue; HPA: bronchus). Unexpected staining may reflect nonspecific binding or endogenous detection activity; compare a reagent control and the expected positive cells on the same run (general IHC practice). |
| Weak, diffuse color across cells and surrounding tissue, without a clear cell specific pattern (general IHC observation). | Diffuse color obscures the selective cytoplasmic pattern needed for interpretation (HPA: tissue IHC). Assess background in a control lacking primary antibody, then review blocking, washes, and detection exposure (general IHC practice). |
| No staining in bone marrow hematopoietic cells or lung macrophages (HPA: High in both). | The run has missed an expected positive population (HPA: bone marrow; HPA: lung). Check that the IHC-validated antibody and detection reagents worked, then review the chosen retrieval and dilution conditions (general IHC practice); this result alone does not identify the failed step. |
💡Expected LCP1 appearanceCall the IHC result positive when expected cells show distinct cytoplasmic staining, including High staining in lung macrophages or bone marrow hematopoietic cells (HPA: tissue IHC); predominant nuclear color or staining of HPA negative cells is suspect (HPA: tissue IHC).
| Cell population and tissue (HPA: tissue IHC) | High staining is reported in lung macrophages, bone marrow hematopoietic cells, lymphoid germinal center cells, spleen red pulp cells, and epididymal glandular cells (HPA: tissue IHC). The epididymal result is a documented exception to a simple immune cell only rule (HPA: epididymis). |
| Compartment and topology (UniProt P13796; HPA: tissue IHC) | UniProt places LCP1 in the cytoskeleton, junctions, and projections and reports no transmembrane segment (UniProt P13796). HPA's tissue IHC profile is selective cytoplasmic expression in immune cells (HPA: tissue IHC); a membrane outline alone should not replace cell type assessment. |
| Antibody evidence (HPA: antibody validation) | HPA019493 and CAB020673 carry Enhanced IHC status, whereas HPA000895 has no stated IHC status in the supplied record (HPA: antibodies). Match interpretation to the antibody actually used; validation for one antibody does not establish identical performance for another (general IHC practice). |
| Isoforms and modifications (UniProt P13796) | UniProt lists two isoforms and multiple modified residues, including phosphorylated sites (UniProt P13796). The supplied record does not map an IHC antibody epitope or show how either feature changes staining, so do not assign a staining difference to them. |
| IF/ICC question: should its membrane signal guide IHC scoring? (HPA: ICC-IF) | HPA ICC-IF supports a main plasma membrane location, with supported actin filament and uncertain cytosol annotations (HPA: ICC-IF). For paraffin section IHC, use the observed selective cytoplasmic tissue pattern as the scoring reference (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known positive cells have no chromogenic signal (HPA: High in bone marrow and lung). | The run may have failed at primary binding, retrieval, or detection; the absent signal does not distinguish among them (general IHC practice). | Verify the positive control, antibody use conditions, retrieval step, and detection reagents before interpreting the test section (general IHC practice). |
| Color is strong in HPA negative cell populations (HPA: adipocytes; bronchial respiratory epithelial cells). | Nonspecific antibody binding or endogenous detection activity can mimic a positive result (general IHC practice). | Compare a control lacking primary antibody and review blocking and detection controls; score the named cell populations separately (general IHC practice). |
| Most staining appears nuclear (HPA: tissue IHC pattern). | This conflicts with reported cytoplasmic and cytoskeletal localization (HPA: tissue IHC; UniProt P13796 subcellular). | Recheck cell identity, compare the positive control, and review antibody concentration and detection background (general IHC practice). |
| A uniform haze hides cell boundaries (general IHC observation). | Excess background prevents a reliable comparison with selective cytoplasmic staining (HPA: tissue IHC). | Inspect the control lacking primary antibody; review wash steps, blocking, and chromogen development (general IHC practice). |
| Only a thin membrane rim is visible in paraffin section IHC (HPA: ICC-IF membrane location). | ICC-IF and tissue IHC report different leading appearances (HPA: ICC-IF; HPA: tissue IHC). | Assess whether expected immune cells also show cytoplasmic staining; use the tissue IHC pattern for the IHC call (HPA: tissue IHC). |
| Epididymal glandular cells stain strongly (HPA: High in epididymis). | This is a reported HPA positive population, despite the broader selective immune cell profile (HPA: epididymis; HPA: tissue IHC). | Record tissue and cell type explicitly, then compare negative cell populations and controls before calling it nonspecific (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing LCP1 staining
A03361 · IHCIHC analysis of LCP1 using anti-LCP1 antibody (A03361). LCP1 was detected in paraffin-embedded section of human B lymphocytic tumor tissue.
A03361 · IHCIHC analysis of LCP1 using anti-LCP1 antibody (A03361). LCP1 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated
A03361 · IFIF analysis of LCP1 using anti-LCP1 antibody (A03361). LCP1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retriev
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136167-LCP1/tissue/cerebral+cortex#imid_5433860
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136167-LCP1/tissue/colon#imid_5433786
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136167-LCP1/tissue/liver#imid_5433779
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136167-LCP1/tissue/kidney#imid_5433788
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019493; green = LCP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019493; green = LCP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LCP1 IHC & IF
🧪Run lymph node first: germinal center cells should stain strongly (HPA: High in lymph node germinal center cells); use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipose tissue adipocytes). On the positive slide, assess morphologically identified cells outside germinal centers as internal negatives only where they lack staining; do not assume the entire surrounding tissue is negative (HPA: High in lymph node germinal center cells).
Positive control tissue: Appendix (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 LCP1 in A-431, U-251MG, U2OS, JURKAT, SiHa, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype-matched rabbit IgG control alongside a target-depleted sample or a peptide-block control when the immunizing peptide is available (hero caption: rabbit anti-LCP1 A03361). For chromogenic detection, block endogenous peroxidase and check endogenous biotin background before using the caption’s biotin-based detection system (hero caption: SABC with DAB); for IF, assess tissue autofluorescence with a no-primary control (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03361 tissue-IHC caption does not state the fixative (hero caption: fixative not stated). Its paraffin-section example uses heat retrieval in citrate buffer at pH 6 for 20 minutes, supporting that retrieval condition for an initial IHC trial without establishing that retrieval is required for every preparation (hero caption: citrate pH 6, 20 minutes). The supplied evidence does not establish whether frozen sections or IF are easier; for lymph node IHC, distinguish germinal center staining from signal due to endogenous peroxidase or biotin in the detection workflow (HPA: High in lymph node germinal center cells; hero caption: SABC with DAB).
HPA tissue IHC evidence for LCP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Appendix | Germinal center cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LCP1 IHC Tips
Troubleshoot LCP1 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, UniProt annotation, and HPA staining patterns.
What retrieval should I try when LCP1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A03361; A03361 tissue-IHC caption). The catalog image used this treatment before overnight incubation at 4°C with 1 μg/ml primary antibody, so compare any retrieval change against those documented conditions (A03361 tissue-IHC caption). If staining remains weak, test a longer heat exposure or an alternative retrieval buffer on adjacent sections, checking whether morphology and background worsen (standard IHC practice). Include a known positive immune-cell compartment and a no-primary control in the same run to distinguish retrieval failure from detection-system background (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or uneven LCP1 staining?
The selected paraffin-section image does not state its fixative, so target-specific fixation sensitivity for LCP1 is unknown (A03361 tissue-IHC caption). Record the specimen’s fixative and fixation duration, then compare similarly processed sections before attributing a weak result to antibody performance (standard IHC practice). On adjacent sections, keep citrate retrieval at pH 6 for 20 minutes and the documented 1 μg/ml primary concentration constant while evaluating staining consistency (A03361 tissue-IHC caption; standard IHC practice). Check section preservation and include a positive immune-cell area, because processing damage and absent target-bearing cells can each make a field appear negative (HPA tissue IHC; standard IHC practice).
Should LCP1 look cytoplasmic or membranous in tissue IHC?
Score the stained cell and its compartment together: HPA reports selective cytoplasmic expression in immune cells in tissue IHC (HPA tissue IHC). LCP1 is an actin-binding protein annotated at the cytoskeleton, cell junctions, projections, and ruffle membranes, with no transmembrane segment (UniProt P13796). Its supported plasma-membrane signal in cell imaging does not establish a uniform membrane-outline pattern for chromogenic tissue sections (HPA subcellular; standard IHC interpretation). If staining appears confined to nuclei or extracellular material, compare nearby positive immune cells, tissue morphology, and a no-primary control before assigning it to LCP1 (HPA tissue IHC; standard IHC practice).
Can this stain distinguish LCP1 isoforms or phosphorylation states?
UniProt lists 2 LCP1 isoforms and multiple modified residues, including phosphoserines at positions 5 and 7 (UniProt P13796). The supplied caption identifies the primary concentration but gives no antibody epitope or isoform-reactivity evidence, so chromogenic intensity cannot be assigned to one isoform or modification state (A03361 tissue-IHC caption; UniProt P13796). Check the antibody’s documented immunogen or epitope before using staining to support a splice-form claim, and use an independently validated reagent if that distinction matters (standard IHC practice). Interpret any shift in staining distribution against cell identity and morphology first, because LCP1 can redistribute to actin-rich projections after phosphorylation (UniProt P13796; standard IHC practice).
How should I assess LCP1 by IF alongside its chromogenic IHC pattern?
Treat IF as a separate assay: the selected antibody evidence here describes chromogenic staining of paraffin sections, with no IF conditions supplied for that antibody (A03361 tissue-IHC caption). Multiplex LCP1 with a validated marker for the expected immune-cell population, and verify each channel with single-stain controls before interpreting overlap (HPA tissue IHC; standard IF practice). Choose fluorophores in channels with low specimen autofluorescence, and inspect an unstained section using the same exposures (standard IF practice). Because LCP1 has no transmembrane segment and occupies cytoskeletal and cytosolic compartments, assess permeabilisation for intracellular epitope access while checking preservation of projections; its precise antibody epitope is unreported here (UniProt P13796; A03361 tissue-IHC caption; standard IF practice).
How do I separate LCP1 staining from diffuse chromogenic background?
First compare the suspect section with a no-primary control, since retained chromogen in that control points toward detection-system or tissue background (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated secondary, a streptavidin-biotin complex, and DAB, so assess blocking and biotin-related background under that detection workflow (A03361 tissue-IHC caption; standard IHC practice). For peroxidase-based DAB detection, verify that the routine endogenous-peroxidase block worked before changing the primary antibody concentration (standard IHC practice). Preserve the documented 1 μg/ml primary condition during the initial comparison, and judge diffuse deposit against HPA’s selective immune-cell cytoplasmic pattern (A03361 tissue-IHC caption; HPA tissue IHC).
How should I quantify LCP1-positive cells across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cell population before scoring, because HPA describes selective immune-cell cytoplasmic expression rather than uniform staining of all cells (HPA tissue IHC). For intensity, report a prespecified H-score from the percentage of eligible cells at each staining grade; for abundance, report positive-cell percentage or positive-cell density per mm² (standard IHC practice). Normalize counts to the same viable tissue area or eligible-cell denominator across sections, excluding folds and necrotic regions by a fixed rule (standard IHC practice). Keep retrieval, development time, counterstain, imaging, and positivity threshold consistent, and report the compartment scored alongside every comparison (standard IHC practice).
What makes an apparent LCP1-positive signal credible rather than artefactual?
A credible pattern includes staining in anatomically identifiable immune cells, consistent with HPA’s selective cytoplasmic tissue profile and high signal in lymph-node germinal-center cells or lung macrophages (HPA tissue IHC). Interpret peripheral or projection-associated staining in cellular context, since LCP1 associates with actin-rich structures and lacks a transmembrane segment (UniProt P13796). Be cautious with isolated nuclear deposits, stained tissue edges, necrotic areas, or broad signal in unrelated cell populations; inspect morphology and the no-primary control before calling those cells positive (HPA tissue IHC; standard IHC practice). DAB precipitate or residual endogenous peroxidase can imitate target staining, so repeat the detection controls when the spatial pattern is inconsistent (standard IHC practice).
Boster reagents
Best LCP1 / Plastin-2 IHC Antibodies
A03361 has pictured LCP1 staining in human paraffin sections and A431 cells; its listed reactivity covers human, mouse, and rat (A03361 IHC/IF captions; catalog reactivity).
Real IHC data
Anti-Plastin L/LCP1 Antibody ®
Cat # A03361
A03361 has IHC images from human B lymphocytic tumor and tonsil paraffin sections, plus an IF/ICC image from A431 cells (A03361 IHC/IF captions). M03361-1 lists IHC and IF/ICC for human, mouse, and rat; M03361-2 lists those applications for human, but neither has an IHC or IF image caption (catalog applications, reactivity, and image_alts).
Which to pick: Choose A03361 for tissue IHC: its own caption documents human paraffin sections, citrate retrieval, and staining at 1 μg/ml; the fixative is unreported (A03361 IHC caption). Choose A03361 for IF/ICC when pictured evidence matters: its A431 image used 2 μg/ml; M03361-1 and M03361-2 are listed rabbit monoclonal alternatives without pictured IHC or IF results (A03361 IF caption; catalog clone, applications, and image_alts). For cross-species work, A03361 and M03361-1 list human, mouse, and rat reactivity, while M03361-2 lists human only; A03361’s pictured samples are human tissue and A431 cells (catalog reactivity; A03361 IHC/IF captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P13796 (PLSL_HUMAN, Plastin-2).
- Human Protein Atlas. LCP1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LCP1 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the cytosol and actin filaments..
- Human Protein Atlas. LCP1 antibody validation summary (3 antibodies).
- IL7R depletion mitigates neuroinflammation and ischemic stroke via AKT dephosphorylation dependent LCP1 suppression in mice. Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics 2026 — PMC13382582.
- LCP1 correlates with immune infiltration: a prognostic marker for triple-negative breast cancer. BMC immunology 2024 — PMC11229261.
- Evaluation of tumorous LCP1 and ADPGK as predictive biomarker for immune-related adverse events in bone and soft tissue sarcomas treated with anti-PD-1 and anti-PD-L1 antibodies. BMC cancer 2025 — PMC11974013.
- Prediction of immune-related adverse events in urological cancer during checkpoint inhibitor immunotherapy through immunohistochemical analysis of tumorous LCP1/ADPGK. Translational oncology 2026 — PMC13266032.
- PubMed PMID:2252891 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057823 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LCP1 (Plastin-2). Expected localisation Cytoplasm in immune cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A03361). Validated IHC antibody: A03361 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human B lymphocytic tumor tissue; fixative not specified (datasheet A03361); 2. Fixation: Image fixative and duration unreported (datasheet A03361); 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 A03361); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03361); 8. Primary antibody: Rabbit anti-LCP1, 0.5-1μg/ml (datasheet A03361); 9. Primary incubation: Overnight at 4 °C (datasheet A03361). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); validate conditions locally.; Caveat: Strong lung macrophage staining can complicate cell scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified; cytoplasmic target (UniProt); Regulation: TCR costimulation shifts LCP1 to synapses (UniProt). Evidence: UniProt P13796, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A03361, PMC11229261, PMC11974013. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LCP1/immunohistochemistry-ihc-guide.