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RILPL2 / RILP-like protein 2 · IHC design guide
Design Immunohistochemistry for RILPL2
Plan paraffin-section RILPL2 IHC using bone marrow hematopoietic cells, which show medium staining, as a positive reference (HPA tissue IHC). The catalog antibody starts at 2.5 μg/mL (datasheet); expect mainly cytoplasmic staining and interpret it cautiously because antibody staining has low consistency with RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RILPL2 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 | Mainly cytoplasmic in most tissues (HPA tissue IHC) | |
| Staining pattern | Mainly cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Esophagus+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) | |
| Caveat | Antibody staining has low consistency with RNA data (HPA tissue IHC) | |
| Regulation | No expression regulator reported (UniProt) | |
| Isoform / epitope | 0 isoforms; one 1–211 chain, with no transmembrane segment (UniProt) |
Section 1
Recommended RILPL2 IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by a published RILPL2 tissue microarray IHC method (PMC11564405).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A14157); 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-RILPL2, 2.5 μg/mL (datasheet A14157) |
| 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 | RILPL2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Mainly cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteTry citrate pH 6.0 HIER at 95–98 °C for 20 min first (page retrieval specification), given RILPL2’s cytosolic localization (UniProt: cytosol).
Section 2
What Is the Expected RILPL2 Staining Pattern?
RILPL2 is a cytosolic protein without a transmembrane segment (UniProt Q969X0). In paraffin-section IHC, expect mainly cytoplasmic staining across many tissues, including medium staining in bone-marrow hematopoietic cells and bronchial respiratory epithelial cells (HPA tissue IHC). Treat this as a provisional pattern: HPA rates the IHC staining Approved, but reports low consistency with RNA expression and says external verification is pending (HPA tissue IHC).
| Cytoplasmic chromogen in bone-marrow hematopoietic cells or bronchial respiratory epithelial cells. | This matches the cell compartments and Medium staining reported for those cells (HPA tissue IHC). Compare signal with neighboring cells and background on the same slide; a matching pattern alone does not resolve HPA's stated RNA–staining inconsistency (HPA tissue IHC). |
| Predominantly nuclear staining, or an apparent cell-surface rim with little cytoplasmic signal. | These compartments diverge from the mainly cytoplasmic tissue pattern (HPA tissue IHC) and cytosolic location (UniProt Q969X0; HPA subcellular). Review morphology and detection controls before interpreting the signal as RILPL2 (general IHC practice). |
| Strong staining in esophageal or vaginal squamous epithelial cells, or skeletal-muscle myocytes. | HPA reports RILPL2 as Not detected in those specific cell populations (HPA tissue IHC). Assess cross-reactivity and endogenous detection activity; the HPA result does not mean every cell in those tissues must be unstained (general IHC practice; HPA tissue IHC). |
| Uniform chromogen across tissue, extracellular spaces, and cells expected to differ. | A diffuse pattern obscures the cell-restricted cytoplasmic interpretation (HPA tissue IHC). Check whether background persists when primary antibody is omitted and inspect blocking, washing, and chromogen development (general IHC practice). |
| No cytoplasmic signal in a selected HPA-positive cell population. | A negative result warrants a technical check, but HPA calls its listed positive populations Medium, not uniformly strong (HPA tissue IHC). Confirm that the relevant cells are present, then review section quality, retrieval, detection, and antibody conditions (general IHC practice). |
💡Expected RILPL2 appearanceCall a positive result when identifiable cells show cytoplasmic chromogen at a credible, cell-resolved intensity—for example, Medium signal in bone-marrow hematopoietic cells (HPA tissue IHC); isolated nuclear staining, diffuse background, or strong signal in HPA-undetected cell populations calls for investigation (HPA tissue IHC; general IHC practice).
| Expected compartment | RILPL2 has no transmembrane segment and is annotated in cytosol, centrosome, and cilium (UniProt Q969X0). Tissue IHC is mainly cytoplasmic (HPA tissue IHC); a possible small ciliary or centrosomal focus should not replace the broader cytoplasmic assessment. |
| Cell-specific tissue evidence | HPA reports Medium staining in adipocytes, adrenal glandular cells, Purkinje cells, and several other listed populations, but Not detected in specified squamous epithelial and muscle cells (HPA tissue IHC). Score the named cell type, not the entire tissue. |
| Validation strength | The listed rabbit polyclonal antibody HPA028958 is IHC Approved, while the tissue profile has low consistency with RNA and awaits external verification (HPA antibodies; HPA tissue IHC). Interpret an unexpected pattern cautiously and use appropriate controls (general IHC practice). |
| Does IF/ICC show the same location? | HPA ICC-IF supports a mainly cytosolic location, with additional vesicle and primary-cilium locations marked Approved (HPA subcellular). Those finer structures can help interpret localization, but this section supplies no IF/ICC protocol (HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells show no chromogen. | The selected section may lack the relevant cells, or the IHC workflow may have failed; HPA's reported signal in those cells is Medium (HPA tissue IHC; general IHC practice). | Locate the named cell population on the counterstained section. Check a concurrently processed positive control, then review retrieval, primary-antibody conditions, and detection according to the antibody's IHC-P instructions (general IHC practice). |
| Signal is mainly nuclear. | This conflicts with cytosolic localization and mainly cytoplasmic tissue staining (UniProt Q969X0; HPA tissue IHC). Nonspecific staining or misread tissue morphology is possible (general IHC practice). | Compare nuclei and cytoplasm against the counterstain, inspect a primary-antibody-omission control, and reassess the result in an HPA-listed positive cell type (general IHC practice; HPA tissue IHC). |
| Strong chromogen appears in an HPA-undetected cell population. | For example, HPA reports esophageal squamous epithelial cells as Not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity can mimic a positive result (general IHC practice). | Verify cell identity and compare with a primary-antibody-omission control. If the detection system uses peroxidase, assess endogenous peroxidase blocking (general IHC practice). |
| Diffuse background makes cell boundaries hard to score. | Nonselective detection, insufficient blocking or washing, or excessive chromogen development can obscure a cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Compare an omission control and a positive section processed together; adjust blocking, washing, and development within the detection system's instructions (general IHC practice). |
| A small punctum is the only apparent positive signal. | Centrosomal or ciliary localization is annotated, but HPA describes tissue IHC as mainly cytoplasmic (UniProt Q969X0; HPA tissue IHC). A punctum alone is hard to assign in chromogenic sections (general IHC practice). | Confirm the structure and cell identity at suitable magnification, then look for reproducible cytoplasmic staining in an HPA-listed positive population (general IHC practice; HPA tissue IHC). |
| Positive and negative populations stain alike. | HPA reports different staining levels for specified cell types, although its tissue pattern has low consistency with RNA expression (HPA tissue IHC). Uniform staining may also reflect background (general IHC practice). | Score each named cell population separately, compare the omission control, and report the discrepancy without treating HPA's Approved rating as independent confirmation of specificity (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing RILPL2 staining
A14157 · IHCImmunohistochemistry of RLP2 in human lung tissue with RLP2 antibody at 2.5 μg/mL.
A14157 · IFImmunofluorescence of RLP2 in human lung tissue with RLP2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000150977-RILPL2/tissue/cerebral+cortex#imid_7880664
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000150977-RILPL2/tissue/colon#imid_7880606
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000150977-RILPL2/tissue/liver#imid_7880607
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000150977-RILPL2/tissue/kidney#imid_7880608
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028958; green = RILPL2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028958; green = RILPL2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RILPL2 IHC & IF
🧪Run adipose tissue first and score staining in adipocytes (HPA: Medium in adipocytes). Use esophagus squamous epithelial cells as the negative tissue (HPA: Not detected); on the adipose slide, score non-adipocyte cells as internal negatives only after confirming they lack RILPL2, with signal at background.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RILPL2 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, using the isotype at the same concentration as the primary antibody (standard IHC practice). Use RILPL2-knockout material as a biological negative where available, and block endogenous peroxidase for chromogenic detection; check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected A14157 lung IHC caption also leaves the fixative unreported (A14157 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier for RILPL2; IF can assess its reported cytosolic and primary-cilium localization with separate validation (HPA subcellular: cytosol supported; primary cilium approved). In paraffin adipose sections, lipid extraction can leave large empty adipocyte spaces and fragile tissue, so assess staining at the cell rim alongside section integrity (standard IHC practice).
HPA tissue IHC evidence for RILPL2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RILPL2 IHC Tips
Troubleshoot RILPL2 staining in paraffin sections by checking retrieval, cellular distribution and controls before comparing signal across samples.
How should I retrieve RILPL2 in paraffin sections when cytoplasmic staining is weak?
Start with citrate pH 6.0 heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule). Allow slides to cool in retrieval buffer, then keep section thickness, heating vessel and development time consistent across the comparison (standard IHC practice). If staining remains weak, compare a small retrieval time series on adjacent sections before trying an alternative buffer, because stronger heating can also raise background or damage morphology (standard IHC practice). Score cytoplasmic signal in the same cell type across conditions; mainly cytoplasmic tissue staining is reported, but its consistency with RNA data is low (HPA: tissue IHC).
Could fixation explain weak or uneven RILPL2 staining in my paraffin sections?
RILPL2 specific sensitivity to fixation is unknown from the supplied evidence; the selected lung image caption does not state its fixative (A14157: tissue IHC caption). Record fixation conditions and processing history for each specimen, and compare sections processed together when troubleshooting uneven staining (standard IHC practice). Inspect morphology and compare staining in well preserved interior tissue with section edges, since uneven processing can complicate interpretation of any chromogenic stain (standard IHC practice). If material permits, test a controlled fixation series with the same retrieval and detection settings, and report the result as an assay finding rather than an established property of RILPL2 (standard IHC practice).
What staining pattern should count as plausible RILPL2 localisation in tissue?
Expect mainly cytoplasmic tissue staining, while treating isolated nuclear signal as a reason to investigate specificity (HPA: tissue IHC; UniProt Q969X0: subcellular location). RILPL2 is also annotated at the centrosome and cilium, and cell imaging reports cytosol with additional vesicle and primary cilium localisation (UniProt Q969X0: subcellular location; HPA: subcellular). Those small structures may be difficult to resolve by routine chromogenic IHC, so do not require visible ciliary puncta to score a cytoplasmic cell as positive (standard IHC practice). Compare staining with morphology and a negative reagent control on the same run, particularly when a punctate or nuclear pattern dominates (standard IHC practice).
How can an unknown antibody epitope affect RILPL2 IHC interpretation?
The supplied record lists no isoforms and gives no antibody epitope, so an isoform specific interpretation is unsupported (UniProt Q969X0: isoforms; A14157: supplied caption). RILPL2 has RH1 at residues 24–106, RH2 at 130–201, and a reported phosphoserine at 107 (UniProt Q969X0: domains and modified residues). Those annotations alone do not establish which sequence the catalog antibody recognizes or whether phosphorylation changes its staining (UniProt Q969X0: annotations; A14157: supplied caption). If epitope mapping becomes available, check whether retrieval conditions and any proposed blocking peptide match that sequence; until then, compare staining with an independent antibody or orthogonal expression evidence (standard IHC practice).
How should I check an RILPL2 IHC result by multiplex immunofluorescence?
Use IF/ICC as a separate validation experiment and pair RILPL2 with a marker for the cell type being scored, using a marker validated for that specimen (standard IF practice). Select spectrally separated fluorophores after checking tissue autofluorescence in an unstained section, and include single stain controls to assess bleed through (standard IF practice). RILPL2 lacks a transmembrane segment and is reported mainly in the cytosol, with additional vesicle and primary cilium localisation, so test gentle permeabilisation to expose intracellular epitopes without losing those structures (UniProt Q969X0: topology; HPA: subcellular; standard IF practice). Do not transfer the catalog lung IHC image concentration of 2.5 µg/mL to IF as a validated setting (A14157: tissue IHC caption).
How do I distinguish diffuse RILPL2 staining from chromogenic background?
Run a no primary control through the same detection and chromogen steps to reveal secondary reagent, endogenous enzyme and tissue pigment contributions (standard IHC practice). For a peroxidase and DAB workflow, use an endogenous peroxidase block and inspect pigment before calling brown cytoplasmic signal positive; these are general workflow controls (standard IHC practice). Titrate the primary antibody and shorten chromogen development if widespread haze obscures cell boundaries, holding retrieval and imaging settings constant during that comparison (standard IHC practice). Mainly cytoplasmic staining across many tissues is reported for RILPL2, but that distribution alone cannot establish that diffuse staining in one section is specific (HPA: tissue IHC; standard IHC practice).
What is a defensible way to quantify RILPL2 chromogenic IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then report the percentage of cytoplasmic positive cells and an intensity based H score using the same thresholds across slides (standard IHC practice; HPA: tissue IHC). If measuring positive cell density, report cells per mm² of viable tissue and exclude folds, tears and necrosis by a prespecified rule (standard IHC practice). Normalise counts to the number of eligible cells or viable tissue area, and compare specimens processed in the same staining batch (standard IHC practice). Keep cytoplasmic and nuclear scores separate; mainly cytoplasmic tissue expression is reported, while HPA notes low consistency between staining and RNA expression (HPA: tissue IHC).
When should an apparent RILPL2 positive tissue section be questioned?
Question signal concentrated only at section edges, folds or necrotic areas, and check whether it persists in a no primary control (standard IHC practice). Predominantly nuclear staining needs further validation because the reported tissue pattern is mainly cytoplasmic and the annotated cellular locations are cytosolic, centrosomal and ciliary (HPA: tissue IHC; UniProt Q969X0: subcellular location). Compare staining in the cell type of interest with reported examples: bronchial respiratory epithelial cells show medium staining, whereas esophageal squamous epithelial cells are listed as not detected (HPA: tissue IHC). Treat those examples as context rather than a universal positive or negative control, because HPA rates the tissue staining Approved but reports low consistency with RNA data and pending external verification (HPA: tissue IHC).
Boster reagents
Best RILPL2 / RILP-like protein 2 IHC Antibodies
A14157 is listed for human IHC-P and IF, with IHC and IF images from human lung tissue (catalog: applications and reactivity; image captions).
Real IHC data
Anti-RLP2 RILPL2 Antibody
Cat # A14157
A14157 has an IHC image of human lung tissue at 2.5 μg/mL (IHC image caption). A14157 also has an IF image of human lung tissue at 20 μg/mL (IF image caption).
Which to pick: For paraffin-section tissue IHC, choose A14157: it is a rabbit antibody listed for human IHC-P, and its own IHC image shows human lung tissue (catalog: host, reactivity and applications; IHC image caption). For IF, choose A14157 based on its human lung image; ICC validation is unreported (catalog: applications; IF image caption). No cross-species option is supported because A14157 lists only human reactivity; its IHC caption does not report a fixative (catalog: reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q969X0 (RIPL2_HUMAN, RILP-like protein 2).
- Human Protein Atlas. RILPL2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RILPL2 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to vesicles and primary cilium..
- Human Protein Atlas. RILPL2 antibody validation summary (1 antibodies).
- RILPL2 as a potential biomarker for predicting enhanced T cell infiltration in non-small cell lung cancer. Immunologic research 2024 — PMC11564405.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:14668488 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RILPL2 (RILP-like protein 2). Expected localisation Mainly cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A14157 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Esophagus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A14157); 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-RILPL2, 2.5 μg/mL (datasheet A14157); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA data (HPA tissue IHC); Isoform / epitope: 0 isoforms; one 1–211 chain, with no transmembrane segment (UniProt); Regulation: No expression regulator reported (UniProt). Evidence: UniProt Q969X0, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to vesicles…, Datasheet A14157, PMC11564405. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RILPL2/immunohistochemistry-ihc-guide.