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LILRB5 / Leukocyte immunoglobulin-like receptor subfamily B member 5 · IHC design guide
Design Immunohistochemistry for LILRB5
Plan LILRB5 chromogenic IHC in paraffin sections using bone marrow hematopoietic cells and spleen red-pulp cells as medium-staining references (HPA tissue IHC). Start the catalog antibody at 1:100–1:300 (datasheet), and treat membrane staining as a molecular expectation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LILRB5 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 | Membrane expected molecularly; tissue compartment unreported (UniProt; HPA tissue IHC) | |
| Staining pattern | Immune-cell subsets in several tissues; compartment unreported (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Bone marrow+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Low antibody–RNA agreement complicates interpretation (HPA tissue IHC) | |
| Regulation | NK-cell expression; induction unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; extracellular versus cytoplasmic epitope needs verification (UniProt) |
Section 1
Recommended LILRB5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by a published LILRB5 IHC protocol using paraffin-embedded human liver cancer tissue (PMC8943947).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet A10961) |
| Fixation | Image fixative and duration unreported (datasheet A10961); 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-LILRB5, 1:100-1:300 (datasheet A10961) |
| 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 | LILRB5-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Distinct expression in subsets of immune cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval setting); the published protocol does not specify retrieval (PMC8943947).
Section 2
What Is the Expected LILRB5 Staining Pattern?
LILRB5 is a membrane receptor with an extracellular region at residues 24–458 and a transmembrane segment at 459–479 (UniProt O75023 topology). In paraffin sections, expect staining in subsets of immune cells, especially bone marrow hematopoietic cells and spleen red pulp cells (HPA tissue IHC). HPA rates the tissue staining “Approved” but reports low consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Discrete staining in some bone marrow hematopoietic cells or spleen red pulp cells. | This matches the reported medium tissue staining; assess the stained cells against neighboring unstained cells (HPA tissue IHC). Membrane localization is expected from receptor topology, although chromogenic sections may not cleanly resolve the cell edge (UniProt O75023 topology; standard IHC practice). |
| Predominantly nuclear staining, with little staining at the cell boundary. | A nuclear pattern conflicts with the membrane annotation and transmembrane topology (HPA subcellular; UniProt O75023 topology). Treat it as suspect and review morphology and detection controls before assigning it to LILRB5 (standard IHC practice). |
| Strong staining in adipocytes or glandular cells while expected immune cell staining is absent. | HPA reports adipocytes in adipose tissue and glandular cells in adrenal gland and appendix as not detected (HPA tissue IHC). Consider nonspecific antibody binding or endogenous detection activity; the staining alone does not identify which cause applies (standard IHC practice). |
| Diffuse color across cells and extracellular areas, without a distinct stained subset. | That distribution is difficult to reconcile with HPA’s distinct immune cell subset pattern (HPA tissue IHC). Review blocking, antibody concentration, washing, and detection controls as general IHC background checks (standard IHC practice). |
| No discernible staining in bone marrow or spleen. | These are reported medium staining contexts, so an absent result warrants a technical check (HPA tissue IHC). It does not by itself prove that the specimen lacks LILRB5; tissue composition and assay performance need review (standard IHC practice). |
💡Expected LILRB5 appearanceCall a result consistent when a subset of bone marrow hematopoietic cells or spleen red pulp cells shows discrete, chiefly membrane-associated staining of approximately medium intensity; broad adipocyte, glandular, or nuclear staining is suspect (HPA tissue IHC; HPA subcellular; UniProt O75023 topology).
| Cell population and tissue choice | Bone marrow hematopoietic cells and spleen red pulp cells are reported at medium, while lymph node and tonsil non-germinal center cells are low (HPA tissue IHC). Compare like cell populations when judging intensity; a whole-section average can obscure a restricted subset (standard IHC practice). |
| Antibody evidence | The listed HPA antibody HPA012069 is a rabbit polyclonal antibody with IHC status “Approved”; no IHC “Enhanced” status is supplied (HPA antibodies). HPA also flags low consistency with RNA expression, so unexpected staining deserves control-based review (HPA tissue IHC reliability). |
| Sequence features and epitope interpretation | The mature chain begins at residue 24, with extracellular and cytoplasmic regions separated by a transmembrane segment (UniProt O75023 processing and topology). The antibody epitope is not supplied; these features cannot establish antigen retrieval requirements or predict staining after fixation. |
| IF/ICC Q: what localization should be expected? | A: Membrane localization is consistent with UniProt topology and HPA’s subcellular summary (UniProt O75023 topology; HPA subcellular). HPA supplies no ICC-IF image-bearing cell lines or main-location annotation here, so there is no image-based IF pattern to compare (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected immune cell staining is absent. | The chosen field may lack the relevant subset, or the IHC run may have failed (HPA tissue IHC; standard IHC practice). | Inspect cell morphology, then check a bone marrow or spleen section known to contain the reported cells and verify the run controls (HPA tissue IHC; standard IHC practice). |
| Only faint signal appears in lymph node or tonsil. | Non-germinal center cells have a reported low staining level in those tissues (HPA tissue IHC). | Judge the relevant cell population rather than requiring the medium intensity reported in bone marrow and spleen (HPA tissue IHC). |
| Adipocytes stain strongly. | Adipocytes are reported as not detected in adipose tissue; nonspecific staining or detection activity is possible (HPA tissue IHC; standard IHC practice). | Compare with a no-primary control and inspect whether any immune cell subset retains a distinct pattern (standard IHC practice; HPA tissue IHC). |
| Brown signal fills the section or follows tissue edges. | Diffuse signal may reflect background from blocking, washing, or detection rather than the reported subset pattern (standard IHC practice; HPA tissue IHC). | Review the no-primary control and detection steps, then reassess discrete staining in morphologically identified cells (standard IHC practice). |
| A strong nuclear pattern dominates. | This conflicts with the supplied membrane localization and transmembrane topology (HPA subcellular; UniProt O75023 topology). | Check controls and tissue morphology before scoring it as LILRB5; seek independent support for any unexpected compartment assignment (standard IHC practice). |
| Protein staining and tissue RNA expectations disagree. | HPA explicitly reports low consistency between antibody staining and RNA expression (HPA tissue IHC reliability). | Report the observed cell-level staining and antibody status separately from RNA expression; avoid using RNA alone to reclassify an IHC-positive or IHC-negative cell (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing LILRB5 staining
A10961 · IHCImmunohistochemical analysis of paraffin-embedded human-spleen, antibody was diluted at 1:100
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105609-LILRB5/tissue/cerebral+cortex#imid_3209000
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105609-LILRB5/tissue/colon#imid_3208934
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105609-LILRB5/tissue/liver#imid_3208935
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000105609-LILRB5/tissue/kidney#imid_3208928
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LILRB5 IHC & IF
🧪Run spleen first: cells in the red pulp should stain at the HPA-reported Medium level, and the selected antibody has a paraffin-section spleen example (HPA: spleen, cells in red pulp, Medium; selected-SKU caption: human spleen). Use bronchus respiratory epithelial cells as the negative tissue (HPA: bronchus, respiratory epithelial cells, Not detected); on the spleen slide, use adjacent cells that lack specific staining as internal negative comparators without assigning an unreported cell type (HPA: spleen, cells in red pulp, Medium).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for LILRB5; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species, immunoglobulin class, and monoclonal or polyclonal format; and, where available, LILRB5 knockout tissue as a biological negative (standard IHC practice). In spleen, assess endogenous peroxidase signal in chromogenic IHC and tissue autofluorescence if evaluating IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU paraffin-section spleen caption does not report a fixative (selected-SKU caption: human spleen, 1:100, fixative unreported). Retrieval dependence is unreported; optimize antigen retrieval and score staining against the control slides, with membrane localisation as the expected pattern (standard IHC practice; UniProt O75023: membrane topology). The supplied evidence does not establish that frozen sections or IF are easier; spleen blood components warrant attention to endogenous peroxidase signal in chromogenic IHC and autofluorescence in IF (standard IHC/IF practice).
HPA tissue IHC evidence for LILRB5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LILRB5 IHC Tips
Troubleshoot LILRB5 staining by checking immune cell context, membrane localisation and controls before interpreting signal intensity (UniProt O75023 topology; HPA tissue IHC).
How should I adjust retrieval when LILRB5 staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes for paraffin section IHC (page retrieval rule: membrane antigen). Run a known positive section alongside the test section and keep section thickness, cooling and detection conditions consistent when comparing results (standard IHC practice). LILRB5 has an extracellular region at residues 24–458 and a cytoplasmic region at 480–590, so epitope accessibility depends on the antibody’s binding site (UniProt O75023 topology). If staining remains weak, compare a second retrieval condition as a fallback on adjacent sections, while watching for tissue damage or increased background (standard IHC practice).
Could fixation explain weak or uneven LILRB5 staining?
Target-specific fixation sensitivity for LILRB5 is unknown from the supplied evidence; the catalog image identifies paraffin-embedded human spleen but does not state its fixative (A10961 tissue-IHC caption). Record the actual fixative and fixation duration for each specimen before comparing staining across cases (standard IHC practice). Process a control section in parallel, then assess preservation, retrieval response and background under the same chromogenic detection conditions (standard IHC practice). If staining varies with fixation history, describe that as an observation from your own sections rather than an established LILRB5 effect (standard IHC practice). The reported 1:100 dilution documents the pictured spleen assay, not fixation tolerance (A10961 tissue-IHC caption).
Which staining compartment and cells should I expect?
Assess LILRB5 primarily as a membrane-associated receptor: its transmembrane segment spans residues 459–479 (UniProt O75023 topology). Membrane-associated staining in immune cell subsets is more consistent with the supplied evidence than uniform staining of an entire tissue compartment (UniProt O75023 subcellular location; HPA tissue IHC profile). HPA reports medium staining in bone marrow hematopoietic cells and spleen red pulp cells, with low staining in lymph node and tonsil non-germinal center cells (HPA tissue IHC). Evaluate morphology and compartment together, because a brown deposit alone does not identify the stained cell or establish receptor specificity (standard IHC interpretation). Compare suspected positive cells with nearby unstained cells on the same section (standard IHC practice).
How can epitope placement change my interpretation of LILRB5 IHC?
Check the antibody’s stated immunogen or epitope before interpreting a negative result, because LILRB5 has three annotated isoforms (UniProt O75023 isoforms). Its extracellular region spans residues 24–458, while residues 480–590 are cytoplasmic; the transmembrane segment lies between them (UniProt O75023 topology). These coordinates guide which side of the receptor an epitope occupies, but they do not establish whether this catalog antibody recognizes every isoform (UniProt O75023 topology and isoforms; epitope not supplied). If epitope information becomes available, compare it with the isoform sequences and document which forms could be detected (standard antibody validation practice). Treat discordant staining between antibodies as a validation question until epitope coverage and controls are resolved (standard IHC practice).
How should I explore LILRB5 in a multiplex IF experiment?
Treat IF/ICC as a separate assay requiring its own validation; the supplied catalog image shows paraffin section chromogenic IHC at 1:100 (A10961 tissue-IHC caption). Pair LILRB5 with a validated marker for the expected immune cell population, then inspect each channel separately before calling colocalisation (HPA tissue IHC profile; standard IF practice). Select a fluorophore and acquisition channel after measuring tissue autofluorescence with an unstained control, and include single-stain controls for spectral overlap (standard IF practice). If the antibody recognizes residues within the cytoplasmic region 480–590, assess permeabilisation; an accessible extracellular epitope may be tested without it (UniProt O75023 topology; standard IF practice). Determine fixation conditions experimentally because no LILRB5-specific IF/ICC fixation evidence is supplied (supplied evidence).
How do I distinguish LILRB5 signal from chromogenic background?
Compare the stained section with a no-primary control and inspect whether brown signal follows cell boundaries or appears as diffuse deposits (standard IHC practice). Include an appropriate peroxidase block before DAB detection, then check whether residual pigment or endogenous enzyme activity persists in control sections (standard chromogenic IHC practice). Because LILRB5 is membrane-associated, widespread nuclear staining would warrant investigation of specificity and detection conditions (UniProt O75023 topology; standard IHC interpretation). Titrate the primary antibody and detection exposure on matched sections while retaining a positive control; 1:100 is the dilution reported for the pictured paraffin-embedded spleen section (A10961 tissue-IHC caption). Score only interpretable cells in preserved areas, avoiding folds and damaged edges (standard IHC practice).
What should I count when comparing LILRB5 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region of interest and cell population before scoring, since HPA describes LILRB5 expression in subsets of immune cells (HPA tissue IHC profile). Report the percentage of positive cells or an H-score from a prespecified intensity scale, and retain representative images for review (standard IHC quantification practice). If cell abundance varies between specimens, report positive-cell density per mm² and normalise positive counts to the number of eligible immune cells in the analysed area (standard IHC quantification practice). Apply the same positivity threshold, section selection and exclusion rules to every case, with observers blinded where feasible (standard IHC quantification practice). Keep staining intensity and cell abundance as separate results so they are not conflated (standard IHC quantification practice).
When is an apparent LILRB5-positive cell convincing?
A convincing result combines staining in an identifiable immune cell with plausible membrane-associated localisation and a clean negative control (HPA tissue IHC profile; UniProt O75023 topology; standard IHC practice). HPA reports medium staining in spleen red pulp cells and bone marrow hematopoietic cells, while its approved tissue profile notes low consistency between antibody staining and RNA expression (HPA tissue IHC). Question nuclear signal, staining confined to section edges or necrotic areas, and deposits that remain after omission of the primary antibody (UniProt O75023 topology; standard IHC interpretation). Check endogenous peroxidase activity before interpreting DAB-positive material as receptor expression (standard chromogenic IHC practice). Describe uncertain cells by morphology and location without assigning a specific lineage from LILRB5 staining alone (standard IHC interpretation).
Boster reagents
Best LILRB5 / Leukocyte immunoglobulin-like receptor subfamily B member 5 IHC Antibodies
A10961 is human-reactive and listed for IHC and IF (catalog: reactivity and applications). Its IHC image shows paraffin-embedded human spleen; no IF image is supplied (catalog: image captions).
Real IHC data
Anti-CD85c LILRB5 Antibody
Cat # A10961
A10961 is listed for human IHC and IF (catalog: reactivity and applications). Its supplied image shows IHC on paraffin-embedded human spleen at 1:100; no IF image is supplied (catalog: image captions).
Which to pick: For paraffin-section IHC, choose A10961: its own image shows human spleen at 1:100, within its listed 1:100–1:300 IHC range (catalog: IHC image caption and dilution). The fixative is unreported in that caption (catalog: IHC image caption). For IF, A10961 is listed at 1:50, but the payload supplies no IF image or ICC validation; no cross-species option is supported because A10961 lists Human reactivity only (catalog: IF dilution, image captions, applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75023 (LIRB5_HUMAN, Leukocyte immunoglobulin-like receptor subfamily B member 5).
- Human Protein Atlas. LILRB5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LILRB5 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. LILRB5 antibody validation summary (1 antibodies).
- Analysis of the expression and prognosis for leukocyte immunoglobulin-like receptor subfamily B in human liver cancer. World journal of surgical oncology 2022 — PMC8943947.
- PubMed PMID:9548455 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LILRB5 (Leukocyte immunoglobulin-like receptor subfamily B member 5). Expected localisation Membrane expected molecularly; tissue compartment unreported (UniProt; HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10961 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human spleen tissue; fixative not specified (datasheet A10961); 2. Fixation: Image fixative and duration unreported (datasheet A10961); 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-LILRB5, 1:100-1:300 (datasheet A10961); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Low antibody–RNA agreement complicates interpretation (HPA tissue IHC); Isoform / epitope: 3 isoforms; extracellular versus cytoplasmic epitope needs verification (UniProt); Regulation: NK-cell expression; induction unreported (UniProt). Evidence: UniProt O75023, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A10961, PMC8943947. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LILRB5/immunohistochemistry-ihc-guide.