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LCP2 / Lymphocyte cytosolic protein 2 · IHC design guide
Design Immunohistochemistry for LCP2
Plan chromogenic IHC for LCP2 in paraffin sections using cytoplasmic staining in lymphoid tissue as the expected pattern (HPA tissue IHC). Use high-staining bone marrow hematopoietic cells or lymph node non-germinal center cells as positive tissue references (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LCP2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in lymphoid tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasm of lymphoid-tissue cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| 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 | Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Cell composition can change staining across lymphoid regions (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | No isoforms; one cytoplasmic chain, no extracellular segment (UniProt) |
Section 1
Recommended LCP2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published LCP2 protocols for lung adenocarcinoma, salivary gland, and placental tissue (PMC8649447; PMC10545362; PMC11979283).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A04880) |
| Fixation | Image fixative and duration unreported (datasheet A04880); 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-LCP2, 1:50-1:200 (datasheet A04880) |
| 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 | LCP2-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval guidance); consider EDTA pH 8.0 for salivary gland sections (PMC10545362).
Section 2
What Is the Expected LCP2 Staining Pattern?
LCP2 should appear predominantly in the cytoplasm of hematopoietic and lymphoid cells, with especially strong staining in bone marrow hematopoietic cells and selected lymphoid tissue compartments (UniProt Q13094: Cytoplasm; HPA tissue IHC: High in these cells). It has no transmembrane segment, so a crisp surface rim is unexpected (UniProt Q13094: topology). HPA rates its tissue IHC pattern Enhanced, reflecting high consistency between antibody staining and RNA expression (HPA tissue IHC: Enhanced).
| Strong cytoplasmic stain in bone marrow hematopoietic cells (HPA tissue IHC: High). | This fits an expected positive control: the signal occupies the compartment assigned by UniProt, in a cell population scored High by HPA (UniProt Q13094: Cytoplasm; HPA tissue IHC: bone marrow). Compare staining with nearby cells on the same section before judging overall intensity. |
| Cytoplasmic stain in lymph node or tonsil non-germinal center cells, or spleen red-pulp cells (HPA tissue IHC: High). | These are supported positive patterns; staining need not be uniform across a lymphoid section (HPA tissue IHC: cell-specific High scores). Assess the annotated cell population, since a tissue-wide score could conceal a correct, restricted pattern. |
| Dominant nuclear stain or a sharp cell-surface rim. | Treat this as a compartment mismatch: UniProt places LCP2 in the cytoplasm and reports no transmembrane segment (UniProt Q13094: subcellular location and topology). Review controls and morphology before interpreting the signal as LCP2; the source record does not support either pattern as expected. |
| Prominent stain in adipocytes or squamous epithelium (HPA tissue IHC: Not detected in listed examples). | Check for cross-reactivity or detection-system activity when these cells stain: HPA reports no signal in adipocytes from adipose tissue or breast, and squamous epithelium from esophagus or oral mucosa (HPA tissue IHC: Not detected). A different cell population in the same specimen may still stain. |
| Diffuse color across cells and extracellular space, or no signal in bone marrow hematopoietic cells. | Diffuse deposition limits compartment scoring; absence in a known-positive population makes a negative result difficult to trust (standard IHC practice; HPA tissue IHC: bone marrow hematopoietic cells High). Check a no-primary control for background and a known-positive section for assay performance before drawing a biological conclusion. |
💡Expected LCP2 appearanceCall a result positive when staining is predominantly cytoplasmic in HPA-supported hematopoietic or lymphoid cell populations, with strong signal possible in bone marrow hematopoietic cells; dominant nuclear or surface-rim staining, or strong signal in an HPA Not detected cell population, warrants control review (UniProt Q13094: Cytoplasm and topology; HPA tissue IHC: High and Not detected cell scores).
| Cell-specific tissue context (HPA tissue IHC). | Use the named cell population when choosing controls: bone marrow hematopoietic cells, lymph node and tonsil non-germinal center cells, and spleen red-pulp cells score High (HPA tissue IHC). A negative score for adipocytes does not make every cell in an adipose-containing specimen negative (HPA tissue IHC: cell-level annotations). |
| Compartment and topology (UniProt Q13094). | LCP2 is cytoplasmic, has no signal peptide or transmembrane segment, and is recorded as one chain spanning residues 1–533 (UniProt Q13094). Those annotations support cytoplasmic interpretation; they do not identify the antibody's epitope or prove a retrieval requirement. |
| Antibody validation (HPA antibodies). | HPA lists three antibodies with Enhanced IHC status: HPA036396, HPA036397 and CAB004574 (HPA antibodies: IHC Enhanced). This supports the reported tissue pattern, but does not establish that an unlisted catalog antibody will give the same staining or specify its working dilution. |
| Antigen retrieval (general IHC practice). | For paraffin IHC, follow the IHC-validated antibody's stated retrieval conditions where available and judge the outcome against positive and no-primary controls (general IHC practice). The supplied UniProt and HPA records do not report LCP2-specific retrieval or fixation sensitivity. |
| IF/ICC Q&A: What pattern should be checked? | Look for cytosolic signal: HPA calls the cytosol its supported main location and lists HEL and serum-starved hTERT-RPE1 among cells with ICC-IF images (HPA subcellular: Cytosol supported). This is an interpretation cue for the separate IF/ICC guide, not an IHC-P protocol option. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in bone marrow hematopoietic cells (HPA tissue IHC: High). | The expected positive control failed; a biological negative cannot be separated from an assay failure on that result alone (HPA tissue IHC: High; general IHC practice). | Confirm the IHC-validated antibody, its stated dilution and retrieval conditions, and that the chromogenic detection reagents worked; repeat alongside a known-positive section (general IHC practice). |
| Weak, patchy signal in an otherwise expected positive population (HPA tissue IHC: High). | Cell selection or section-to-section assay variation may account for the appearance; HPA's cell-specific score does not set an intensity threshold for every specimen (HPA tissue IHC: cell annotations; general IHC practice). | Compare the same annotated cell population in a known-positive section, then review the antibody's IHC-P conditions and counterstain so faint cytoplasmic signal can be judged (general IHC practice). |
| Predominantly nuclear stain or a continuous membrane rim. | The compartment conflicts with cytoplasmic LCP2 and its lack of a transmembrane segment (UniProt Q13094: location and topology). | Check staining without primary antibody, inspect cell boundaries and nuclei, and seek a concordant cytoplasmic pattern with an independently validated antibody if available (general IHC practice; HPA antibodies: Enhanced IHC entries). |
| Strong stain in adipocytes or listed squamous epithelial cells (HPA tissue IHC: Not detected). | Possible nonspecific antibody binding or detection activity; the HPA negative calls apply to those specific cell populations (HPA tissue IHC: Not detected; general IHC practice). | Compare a no-primary control and an HPA-supported positive cell population, then reassess the IHC-validated antibody and detection controls before assigning LCP2 expression (general IHC practice; HPA tissue IHC). |
| Diffuse chromogen obscures cell borders. | Background deposition prevents a reliable cytoplasmic call; it does not establish LCP2 expression (general IHC practice; UniProt Q13094: Cytoplasm). | Inspect a no-primary control and review blocking, wash steps and detection conditions; score only cells whose signal can be localized clearly (general IHC practice). |
| Moderate glandular or lung macrophage staining seems inconsistent with a lymphoid-focused pattern. | HPA also reports Medium staining in adrenal and gallbladder glandular cells and lung macrophages (HPA tissue IHC: Medium); a non-lymphoid site alone does not establish cross-reactivity. | Identify the stained cell type and compartment first, compare the relevant HPA cell-level annotation, and use controls if the pattern remains discordant (HPA tissue IHC: Medium; UniProt Q13094: Cytoplasm; general IHC practice). |
Experiment evidence
IHC & IF images showing LCP2 staining
A04880 · IHCImmunohistochemistry (IHC) analyzes of SLP-76 (G122) pAb in paraffin-embedded human breast carcinoma tissue at 1:100.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000043462-LCP2/tissue/cerebral+cortex#imid_10339849
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000043462-LCP2/tissue/colon#imid_10339775
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000043462-LCP2/tissue/liver#imid_10339776
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000043462-LCP2/tissue/kidney#imid_10339769
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA036396; green = LCP2, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA036396; green = LCP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LCP2 IHC & IF
🧪Run spleen first: cells in the red pulp should stain (HPA: High in spleen red-pulp cells). Run adipose tissue as the negative, where adipocytes should remain unstained (HPA: Not detected in adipose-tissue adipocytes); on the spleen slide, assess nonhematopoietic stromal cells as internal negative cells (UniProt Q13094: high expression in leukocytes; not detected in fibroblast cell lines).
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 LCP2 in HEL, hTERT-RPE1 (serum starved), with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a control using an antibody matched to the primary antibody’s host species and immunoglobulin class, and LCP2-knockout material or a peptide-block control if a matching immunogen is available (standard IHC practice). Block endogenous peroxidase when using chromogenic detection in spleen red pulp; for IF, check red-pulp autofluorescence with an unstained section (standard IHC/IF practice).
⚠️Feasibility: The supplied evidence reports no LCP2-specific fixation window or fixation effect, and the fixative for the selected A04880 paraffin-section caption is unreported (A04880 tissue-IHC caption). Antigen-retrieval dependence is unreported, so optimize retrieval on the paraffin positive control; the evidence does not establish whether frozen sections or IF are easier (A04880 tissue-IHC caption; HPA: cytosolic ICC-IF localization). Blood-rich spleen red pulp warrants attention to endogenous peroxidase in chromogenic IHC and autofluorescence in IF (standard IHC/IF practice).
HPA tissue IHC evidence for LCP2
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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LCP2 IHC Tips
Troubleshoot LCP2 staining by checking cytoplasmic localisation, immune cell identity and matched controls (UniProt Q13094; HPA tissue IHC).
What retrieval conditions should I try when LCP2 staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (this page’s IHC retrieval setting). Let sections cool in buffer, then compare them with a matched section processed without retrieval so that any gain in signal can be judged against background (standard IHC practice). The selected A04880 image shows staining in a paraffin section at 1:100, but its caption does not report a retrieval method (A04880 tissue-IHC caption). If staining remains weak, adjust heating time on matched sections while holding antibody concentration and detection constant; assess cytoplasmic signal in lymphoid cells alongside tissue damage (standard IHC practice; HPA tissue IHC).
Could fixation explain weak LCP2 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the paraffin-section caption for A04880 does not state a fixative (A04880 tissue-IHC caption). Record the fixative, fixation duration and processing history for each block before comparing staining, because those conditions can affect antigen accessibility in IHC (standard IHC practice). Compare similarly processed sections with the same retrieval and detection settings, using lymphoid tissue as a reference for expected cytoplasmic staining (standard IHC practice; HPA tissue IHC). If one processing batch stains poorly, optimise retrieval on matched sections before attributing the difference to LCP2 abundance; the available evidence does not establish a fixation effect for this target (this page’s IHC retrieval setting; A04880 tissue-IHC caption).
Where should convincing LCP2 staining appear in tissue?
Look for predominantly cytoplasmic staining in appropriate cells: LCP2 is annotated in the cytoplasm, and tissue IHC describes cytoplasmic expression in lymphoid tissues (UniProt Q13094 localisation; HPA tissue IHC). High staining is reported in bone marrow hematopoietic cells, lymph node non-germinal center cells and spleen red-pulp cells (HPA tissue IHC). Because LCP2 has no transmembrane segment, an exclusively crisp membrane outline needs further scrutiny rather than immediate acceptance as its expected pattern (UniProt Q13094 topology). Compare morphology and a matched immune cell marker where cell identity is uncertain; isolated nuclear staining should prompt review of controls and counterstain (standard IHC practice; UniProt Q13094 localisation).
How should epitope location guide troubleshooting of inconsistent LCP2 staining?
The supplied record lists one 533-aa LCP2 chain and 0 isoforms, so it provides no annotated isoform pattern to explain discordant IHC staining (UniProt Q13094 processing and isoforms). LCP2 contains a SAM domain at residues 15–81, an SH2 domain at 422–530, and documented phosphorylation sites including Tyr23 and Ser207 (UniProt Q13094 domains and modified residues). Check the catalog antibody’s documented immunogen or epitope before interpreting differences between antibodies; its epitope is not supplied here (provided A04880 caption). Compare matched sections under the same retrieval conditions and report any epitope information with the staining result, since epitope accessibility can vary with processing (standard IHC practice).
How can IF help check a puzzling chromogenic LCP2 pattern?
Use IF as a complementary localisation check, while judging this page’s primary result on the chromogenic paraffin sections (standard IHC/IF practice; A04880 tissue-IHC caption). Multiplex LCP2 with a marker for the suspected immune cell population, and include single-stain controls to distinguish overlapping cells from apparent colocalisation (standard IF practice; UniProt Q13094 function). Choose a fluorophore channel with low background in the tissue under study, then inspect unstained tissue for autofluorescence before interpreting faint signal (standard IF practice). Because LCP2 is cytosolic and has no transmembrane segment, permeabilise fixed cells for IF access to the intracellular epitope and verify the resulting cytosolic pattern (HPA subcellular; UniProt Q13094 topology; standard IF practice).
How do I reduce diffuse or nonspecific chromogenic signal?
First compare a no-primary control with the stained section; residual color can expose endogenous enzyme activity or nonspecific detection in chromogenic IHC (standard IHC practice). For a peroxidase and DAB workflow, apply a peroxidase block, use appropriate protein blocking, and titrate the primary antibody while keeping retrieval and development time consistent (standard IHC practice). The A04880 paraffin-section image used 1:100, which is a documented image condition rather than an established optimum for every specimen (A04880 tissue-IHC caption). Assess whether improved contrast preserves cytoplasmic staining in expected lymphoid cells, since widespread color in unrelated structures weakens the biological interpretation (HPA tissue IHC; UniProt Q13094 localisation).
What should I measure when LCP2-positive cells vary across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, then report the percentage of cytoplasm-positive cells and staining intensity within that population (standard IHC practice; UniProt Q13094 localisation). An H-score can combine intensity with percentage, while positive-cell density per mm² can describe infiltrates when tissue composition varies (standard IHC quantification practice). Normalise cell counts to the measured viable tissue area, and compare staining percentages against the number of eligible cells rather than all cells in the section (standard IHC quantification practice). Apply the same threshold, retrieval, detection and sampling rules across groups; record immune cell composition because LCP2 is prominent in lymphoid tissues and leukocytes (standard IHC practice; HPA tissue IHC; UniProt Q13094 tissue specificity).
How can I distinguish true LCP2 staining from artefact?
A convincing result combines cytoplasmic signal with the expected cell context, such as lymphoid tissue or bone marrow hematopoietic cells (UniProt Q13094 localisation; HPA tissue IHC). Scrutinise strong epithelial or isolated nuclear color against tissue morphology and matched controls, since those patterns do not match the reported cytoplasmic lymphoid profile (HPA tissue IHC; standard IHC practice). Exclude section edges and necrotic regions from interpretation when they show disproportionate staining, and check a no-primary section for color caused by the detection workflow (standard IHC practice). The A04880 breast carcinoma image establishes staining in a paraffin specimen at 1:100; assess which cells stain before assigning that signal to carcinoma cells (A04880 tissue-IHC caption; standard IHC practice).
Boster reagents
Best LCP2 / Lymphocyte cytosolic protein 2 IHC Antibodies
The catalog antibody has real IHC data from paraffin-embedded human breast carcinoma (image caption). Human, mouse, and rat reactivity is listed; no IF application or image is supplied (catalog).
Real IHC data
Anti-SLP-76 (G122) LCP2 Antibody
Cat # A04880
A04880 has an IHC image from paraffin-embedded human breast carcinoma at 1:100 (image caption). Its listed applications are IHC and WB, and its listed reactivity is human, mouse, and rat (catalog).
Which to pick: Choose A04880 for paraffin-section tissue IHC because its own image shows that preparation; the fixative is unreported (image caption). No SKU in the payload lists IF/ICC or shows an IF image (catalog). A04880 is the cross-species candidate because human, mouse, and rat reactivity is listed, though its IHC image shows only human tissue (catalog; image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13094 (LCP2_HUMAN, Lymphocyte cytosolic protein 2).
- Human Protein Atlas. LCP2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LCP2 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. LCP2 antibody validation summary (3 antibodies).
- Lymphocyte cytosolic protein 2 is a novel prognostic marker in lung adenocarcinoma. The Journal of international medical research 2021 — PMC8649447.
- A novel prognostic biomarker LCP2 correlates with metastatic melanoma-infiltrating CD8(+) T cells. Scientific reports 2021 — PMC8080722.
- Identification of key genes in salivary gland in Sjögren's syndrome complicated with Hashimoto thyroiditis: Common pathogenesis and potential diagnostic markers. Medicine 2023 — PMC10545362.
- Using weighted gene co-expression network analysis to identify key genes related to preeclampsia. Frontiers in immunology 2025 — PMC11979283.
- PubMed PMID:7706237 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LCP2 (Lymphocyte cytosolic protein 2). Expected localisation Cytoplasmic staining in lymphoid tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04880 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A04880); 2. Fixation: Image fixative and duration unreported (datasheet A04880); 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-LCP2, 1:50-1:200 (datasheet A04880); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Cell composition can change staining across lymphoid regions (HPA tissue IHC); Isoform / epitope: No isoforms; one cytoplasmic chain, no extracellular segment (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt Q13094, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet A04880, PMC8649447, PMC10545362. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LCP2/immunohistochemistry-ihc-guide.