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LRBA / Lipopolysaccharide-responsive and beige-like anchor protein · IHC design guide
Design Immunohistochemistry for LRBA
Plan LRBA staining in paraffin sections using the IHC-validated antibody A04947 (datasheet A04947). Interpret cytoplasmic staining against the variable tissue profile reported by HPA (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LRBA 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 across tissues (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04947) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression varies across tissues (HPA tissue IHC) | |
| Isoform / epitope | Two isoforms; check epitope coverage when comparing staining (UniProt) |
Section 1
Recommended LRBA IHC & IF Protocols
The catalog antibody protocol uses Tris-EDTA pH 9.0 retrieval (datasheet A04947). Two published LRBA IHC protocols provide additional tissue-specific examples (PMC12815959; PMC12266018).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A04947) |
| Fixation | Image fixative and duration unreported (datasheet A04947); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A04947); 20 min, 95–100 °C (standard) |
| 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-LRBA, 1:50 recommended; image 1:200 (datasheet A04947) |
| 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 | LRBA-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues at variable levels. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 9.0 retrieval (datasheet A04947); the published heat-retrieval protocol does not specify a buffer (PMC12815959).
Section 2
What Is the Expected LRBA Staining Pattern?
LRBA should appear as variable cytoplasmic staining across tissues, with strong staining reported in several epithelial cell populations (HPA tissue IHC). Its membrane associations include the endoplasmic reticulum, trans-Golgi network, lysosome and cell membrane; UniProt annotates one transmembrane segment (UniProt P50851 topology). HPA rates the tissue IHC profile Approved, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Strong cytoplasmic staining in bronchial respiratory epithelium or colon glandular cells (HPA tissue IHC). | This matches two reported High cell populations (HPA tissue IHC). Judge the stained cell population and cytoplasmic compartment together; HPA reports variable expression across tissues (HPA tissue IHC). A positive field alone does not establish antibody specificity (standard IHC practice). |
| Predominantly nuclear staining, with little cytoplasmic signal, in a candidate positive section. | A nuclear-dominant pattern conflicts with HPA's cytoplasmic tissue profile and UniProt's membrane locations (HPA tissue IHC; UniProt P50851). Treat it as suspect; review controls and staining conditions before scoring the nuclei as LRBA positive (standard IHC practice). |
| The strongest signal is in cells outside the expected stained population for the selected tissue. | For example, HPA reports High staining in colon glandular cells (HPA tissue IHC). Unexpected staining elsewhere may reflect cross-reactivity or endogenous detection activity (standard IHC practice). Compare cell identity, compartment and negative controls before assigning it to LRBA. |
| A broad brown haze covers tissue and background, obscuring cell boundaries. | That distribution cannot be read as the variable cellular cytoplasmic pattern reported for LRBA (HPA tissue IHC). Background can arise from nonspecific binding or chromogenic detection activity (standard IHC practice); assess the matched negative control before changing the interpretation. |
| No cytoplasmic staining appears in bronchial respiratory epithelial cells or colon glandular cells. | Both are reported High by HPA, so an absent signal warrants a technical check (HPA tissue IHC). HPA's Approved rating has medium staining–RNA consistency and pending external verification; one negative section does not by itself establish that LRBA is absent (HPA tissue IHC). |
💡Expected LRBA appearanceCall a section positive when the appropriate epithelial cells show clear, potentially strong cytoplasmic staining (HPA tissue IHC); nuclear-dominant staining or uniform haze is suspect (HPA tissue IHC; standard IHC practice).
| Tissue and cell population | HPA reports High staining in appendix, colon and duodenum glandular cells and bronchial respiratory epithelium, but Low staining in glia and fibroblasts (HPA tissue IHC). Select and interpret controls by cell population, since whole-section intensity can obscure that distinction (standard IHC practice). |
| Compartment and image resolution | HPA tissue IHC describes cytoplasmic staining; HPA ICC-IF supports a mainly Golgi location, with additional cytosol localization marked uncertain (HPA tissue IHC; HPA subcellular). A chromogenic section need not resolve individual organelle membranes to fit that profile (standard IHC practice). |
| Topology and epitope location | UniProt annotates one transmembrane segment at residues 1531–1548 and membrane locations including the trans-Golgi network and lysosome (UniProt P50851 topology). The supplied record gives no antibody epitope, so topology cannot predict whether a particular retrieval condition exposes it. |
| Isoforms and processing | UniProt lists 2 isoforms and a chain spanning residues 2–2863, with no signal peptide or propeptide annotation (UniProt P50851). Without an antibody epitope map, these annotations cannot establish which isoforms the IHC-validated antibody detects. |
| Antibody validation scope | HPA lists HPA019366 and HPA023597 as IHC Approved; HPA019366 is also ICC Supported (HPA antibodies). The tissue profile has medium staining–RNA consistency and awaits external verification (HPA tissue IHC). These ratings support a working expectation, not an absolute pass rule for every specimen. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported High cell population has no IHC signal (HPA tissue IHC). | The section, detection run or antibody conditions may have failed (standard IHC practice); HPA's High designation does not guarantee every specimen will stain (HPA tissue IHC). | Run a known-positive section in the same batch; confirm the antibody's IHC-P instructions, retrieval and detection steps, then inspect the intended cells before calling the sample negative (standard IHC practice). |
| Signal is nuclear-dominant despite an otherwise clean section. | Nuclear dominance does not match the reported cytoplasmic tissue pattern or annotated membrane locations (HPA tissue IHC; UniProt P50851). | Check the negative control, antibody concentration and staining distribution; require convincing cytoplasmic signal in a reported positive cell population before interpreting the nuclear signal as LRBA (standard IHC practice; HPA tissue IHC). |
| Brown staining appears in unexpected cells or persists in a negative control. | Cross-reactivity, endogenous enzyme activity or nonspecific detection may contribute (standard IHC practice); the location conflicts with the selected tissue's reported cell pattern (HPA tissue IHC). | Compare a matched no-primary control and the known-positive cell population. Check the detection system's endogenous-activity blocking and background controls before scoring unexpected cells (standard IHC practice). |
| Diffuse background makes cytoplasmic boundaries hard to judge. | Nonspecific binding or excessive detection signal can obscure cellular staining (standard IHC practice). The resulting haze cannot establish the HPA cytoplasmic pattern (HPA tissue IHC). | Review the negative control and the catalog antibody's IHC-P working conditions; adjust blocking, antibody dilution or detection time as appropriate to the validated workflow, then reassess cell-specific signal (standard IHC practice). |
| A Low-reference cell population stains faintly, or a High-reference population varies between sections. | HPA reports variable cytoplasmic levels across tissues, including Low staining in glia and fibroblasts and High staining in several epithelia (HPA tissue IHC). Its tissue profile has medium staining–RNA consistency (HPA tissue IHC). | Score the named cell population and compartment rather than whole-section color; compare sections processed together and include a reported High population as a reference (standard IHC practice; HPA tissue IHC). |
| IF/ICC: what localization should an independently designed experiment seek? | HPA supports a mainly Golgi location and marks additional cytosol localization uncertain; HPA019366 has ICC Supported status (HPA subcellular; HPA antibodies). | Use Golgi-enriched signal as the principal localization expectation, with the cytosolic component interpreted cautiously (HPA subcellular). Consult the separate IF/ICC guide for experiment design. |
Experiment evidence
IHC & IF images showing LRBA staining
A04947 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198589-LRBA/tissue/cerebral+cortex#imid_6045880
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198589-LRBA/tissue/colon#imid_6045806
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198589-LRBA/tissue/liver#imid_6045791
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198589-LRBA/tissue/kidney#imid_6045792
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019366; green = LRBA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019366; green = LRBA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LRBA IHC & IF
🧪Run appendix first and expect glandular cells to stain (HPA: High in appendix glandular cells). HPA detects LRBA in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; cells without chromogen on the appendix slide should show counterstain only, but are not validated LRBA-negative cells (HPA: no negative tissue rows; standard IHC control practice).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: LRBA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LRBA in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a species- and clonality-matched isotype control; where available, a matched LRBA knockout specimen provides a biological negative (standard IHC control practice). Block endogenous peroxidase for chromogenic detection in appendix, and assess endogenous biotin if using avidin–biotin detection (standard IHC control practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A04947 paraffin-section caption does not state a fixative (selected A04947 tissue-IHC caption). Start retrieval optimization with Tris-EDTA at pH 9.0, the reported condition, without assuming retrieval is required for every specimen (selected A04947 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; check appendix glandular lumina for nonspecific chromogen when scoring epithelial staining (HPA: High in appendix glandular cells; standard IHC control practice).
HPA tissue IHC evidence for LRBA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: LRBA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced LRBA IHC Tips
Troubleshoot LRBA chromogenic IHC in paraffin sections using the A04947 tonsil caption, the tissue staining profile, and known protein localisation.
Which retrieval conditions should I try first for weak LRBA staining?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 for paraffin sections (datasheet A04947). The A04947 human tonsil image used this buffer with primary antibody at 1:200 overnight at 4°C (datasheet A04947). If staining is weak, check that heating and cooling were consistent across slides before adjusting retrieval time as a controlled optimization (standard IHC practice). Compare changes against a tonsil section processed in parallel, because tonsil is the demonstrated tissue for this antibody (datasheet A04947). Record tissue morphology alongside signal: excessive heat can damage sections and make stronger colour difficult to interpret (standard IHC practice).
Could fixation explain weak or uneven LRBA staining?
The A04947 image documents paraffin-embedded human tonsil, but does not state its fixative (datasheet A04947). LRBA-specific sensitivity to any fixation method or duration is therefore unknown from the supplied evidence (datasheet A04947). When investigating weak staining, record the fixative, fixation duration, section thickness, and processing history for each specimen (standard IHC practice). Process a tonsil reference beside the study sections using Tris-EDTA at pH 9.0 and the documented 1:200 primary dilution (datasheet A04947). If the reference stains and the study sections do not, examine specimen processing and tissue preservation before drawing a biological conclusion (standard IHC practice).
What staining pattern is plausible for LRBA in tissue sections?
Assess LRBA primarily as cytoplasmic staining across tissue cell populations, with intensity varying by tissue (HPA tissue IHC: cytoplasmic expression in all tissues). A perinuclear or punctate component can be biologically plausible because LRBA is supported at the Golgi apparatus in cell imaging (HPA subcellular: Golgi supported). The protein is also annotated at endoplasmic reticulum, trans-Golgi network, lysosome, and cell membranes (UniProt P50851 localisation). Its one annotated transmembrane segment spans residues 1531–1548 (UniProt P50851 topology). Use cell morphology and a matched reference when judging these patterns; diffuse nuclear-only colour conflicts with the supplied localisation evidence (HPA subcellular; UniProt P50851 localisation).
How should epitope location affect my LRBA IHC interpretation?
LRBA has 2 annotated isoforms, so establish which sequence the catalog antibody recognizes before treating every staining difference as altered protein abundance (UniProt P50851 isoforms). The supplied caption gives an IHC dilution and retrieval method but no antibody epitope coordinates (datasheet A04947). Compare the stated epitope, if available, with the 1531–1548 transmembrane segment and the BEACH-type PH and BEACH domains (UniProt P50851 topology and domains). Document whether a tested antibody could recognize both isoforms; the supplied evidence does not establish that coverage (UniProt P50851 isoforms; datasheet A04947). Interpret altered staining cautiously until epitope accessibility under Tris-EDTA pH 9.0 retrieval is checked experimentally (datasheet A04947; standard IHC practice).
How can IF help check an ambiguous LRBA IHC pattern?
Use IF as a separate localisation check when chromogenic IHC cannot resolve a perinuclear signal; Golgi localisation is supported in cell imaging (HPA subcellular). Multiplex LRBA with a marker that identifies the expected cell population in the specimen, then assess overlap within individual cells (standard IF practice). Choose fluorophores and imaging channels after measuring tissue autofluorescence, and include single-colour controls to identify bleed-through (standard IF practice). Plan permeabilisation around the antibody epitope’s side of LRBA’s 1531–1548 membrane-spanning region; its side is not specified here (UniProt P50851 topology). Keep IF fixation and antibody conditions independently validated, since the A04947 caption supplies paraffin tissue IHC conditions only (datasheet A04947).
How do I reduce diffuse or patchy background in LRBA IHC?
First compare a no-primary control with the stained section to identify colour produced by detection reagents or tissue (standard IHC practice). Include an endogenous peroxidase block before chromogenic detection and assess nonspecific secondary binding with the appropriate control (standard IHC practice). The documented A04947 tonsil workflow used primary antibody at 1:200 overnight at 4°C and secondary antibody at 1:200 for 45 minutes at room temperature (datasheet A04947). If background rises with staining intensity, titrate the primary around the documented condition while keeping retrieval and development consistent (datasheet A04947; standard IHC practice). Check section edges and damaged areas separately because their colour can distort interpretation of cellular staining (standard IHC practice).
What is a defensible way to quantify LRBA IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, then report the percentage of positive cells and intensity by compartment (standard IHC practice). An H-score can combine intensity categories with their positive-cell percentages; keep thresholds and illumination consistent across the slide set (standard IHC practice). For discrete positive cells, density per mm² can supplement the score when cell counts matter (standard IHC practice). Normalize scores to the number of evaluable target cells or the analysed tissue area, and exclude folds, necrosis, and section edges (standard IHC practice). Stratify by cell type because HPA reports variable cytoplasmic LRBA staining across tissues and cell populations (HPA tissue IHC).
When should an apparent LRBA-positive result be questioned?
A credible result should show interpretable cellular staining that fits LRBA’s reported cytoplasmic tissue profile and supported Golgi localisation (HPA tissue IHC; HPA subcellular). High staining in glandular cells from colon or appendix can serve as a contextual comparison, not a universal threshold (HPA tissue IHC: high in colon and appendix glandular cells). Question nuclear-only colour, signal restricted to cut edges or necrotic areas, and colour reproduced in a no-primary control (HPA subcellular; standard IHC practice). Check endogenous peroxidase activity when interpreting chromogenic colour, especially if the control also develops signal (standard IHC practice). HPA rates tissue staining Approved with medium RNA–staining consistency and pending external verification, so corroborate consequential findings independently (HPA tissue IHC reliability).
Boster reagents
Best LRBA / Lipopolysaccharide-responsive and beige-like anchor protein IHC Antibodies
A04947 has a real IHC image from paraffin-embedded human tonsil (A04947 image caption). IF is listed for human and mouse, but no IF image is supplied (A04947 catalog: applications, reactivity, IF image alts).
Real IHC data
Anti-LRBA Antibody
Cat # A04947
A04947 will render with its paraffin-embedded human tonsil IHC image (A04947 image caption). The catalog lists IHC and IF applications and human and mouse reactivity; it supplies no IF image (A04947 catalog: applications, reactivity, IF image alts).
Which to pick: For tissue IHC, choose A04947: its own image caption documents staining in paraffin-embedded human tonsil, using 1:200 primary antibody overnight at 4°C and Tris-EDTA retrieval at pH 9.0 (A04947 image caption). For IF, A04947 is the listed option; ICC is not listed, and no IF image is supplied (A04947 catalog: applications, IF image alts). A04947 lists human and mouse reactivity, but the pictured tissue is human; its host is rabbit, clonality is unreported, and the tissue fixative is unreported (A04947 catalog: host, clone, reactivity; A04947 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50851 (LRBA_HUMAN, Lipopolysaccharide-responsive and beige-like anchor protein).
- Human Protein Atlas. LRBA tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LRBA subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the cytosol..
- Human Protein Atlas. LRBA antibody validation summary (2 antibodies).
- Targeting LRBA triggers CTLA4 degradation and antitumor immunity for cancer immunotherapy. Nature communications 2025 — PMC12815959.
- A precision medicine approach to primary immunodeficiency disease: Ataluren strikes nonsense mutations once again. Molecular therapy : the journal of the American Society of Gene Therapy 2025 — PMC12266018.
- PubMed PMID:11254716 — UniProt-cited evidence.
- PubMed PMID:12160729 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LRBA (Lipopolysaccharide-responsive and beige-like anchor protein). Expected localisation Cytoplasmic staining across tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04947). Validated IHC antibody: A04947 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A04947); 2. Fixation: Image fixative and duration unreported (datasheet A04947); 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: Tris-EDTA pH 9.0 (datasheet A04947); 20 min, 95–100 °C (standard); 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-LRBA, 1:50 recommended; image 1:200 (datasheet A04947); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: Two isoforms; check epitope coverage when comparing staining (UniProt); Regulation: Expression varies across tissues (HPA tissue IHC). Evidence: UniProt P50851, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A04947, PMC12815959, PMC12266018. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LRBA/immunohistochemistry-ihc-guide.