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REEP2 / Receptor expression-enhancing protein 2 · IHC design guide
Design Immunohistochemistry for REEP2
Plan REEP2 chromogenic IHC in paraffin sections using cytoplasmic neuronal staining as the expected tissue pattern (HPA tissue IHC). The guide covers controls, antibody dilution, detection and scoring, with high staining in caudate neurons and cerebellar Purkinje cells as reference patterns (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
REEP2 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 | Neuronal cytoplasm (HPA tissue IHC); membrane protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in neurons (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Adrenal gland+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 | Presumed off-target staining was observed (HPA tissue IHC) | |
| Regulation | Staining regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope side affects access; map unknown (UniProt) |
Section 1
Recommended REEP2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published REEP2 paraffin-section protocol (PMC10978033).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A11618); 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-REEP2, 5 μg/mL (datasheet A11618) |
| 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 | REEP2-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in neurons. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); the published method uses microwave sodium citrate for 10 min (PMC10978033).
Section 2
What Is the Expected REEP2 Staining Pattern?
REEP2 is a membrane protein with 2 transmembrane segments (UniProt Q9BRK0 topology). In paraffin sections, expect cytoplasmic staining in neurons, particularly in caudate and cerebral cortex neurons and cerebellar Purkinje cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while noting medium agreement with RNA data and presumed off-target binding that was disregarded (HPA tissue IHC).
| Cytoplasmic staining in caudate or cortical neurons, or cerebellar Purkinje cells (HPA tissue IHC). | This matches the reported positive pattern; these cell types stain at a High level (HPA tissue IHC). Compare stained cells with neighboring cells on the same section when judging whether the pattern is selective (general IHC practice). |
| Predominantly nuclear staining in cells otherwise expected to be positive. | Nuclear localization does not match HPA's cytoplasmic tissue pattern or approved ER localization in ICC-IF (HPA tissue IHC; HPA subcellular). Treat it as questionable and check whether it persists with the IHC-validated antibody under its documented conditions (general IHC practice). |
| Strong staining in glandular cells of adrenal gland, appendix, or duodenum. | Those cell populations are reported as Not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity before calling the signal REEP2; HPA also reports presumed off-target binding in its tissue assessment (HPA tissue IHC; general IHC practice). |
| Broad, diffuse color across cells and tissue compartments, obscuring cell boundaries. | That distribution cannot establish the reported neuron-selective cytoplasmic pattern (HPA tissue IHC). Evaluate background with an appropriate negative detection control and review blocking, washing, and detection conditions (general IHC practice). |
| No detectable staining in caudate neurons or cerebellar Purkinje cells. | These are reported High staining populations (HPA tissue IHC), so a blank result warrants a technical check. It does not alone show that REEP2 is absent: section quality, antibody conditions, and detection performance also affect IHC (general IHC practice). |
💡Expected REEP2 appearanceCall a section positive when neurons show distinct cytoplasmic staining, strongest in reported High populations; diffuse tissue-wide, nuclear, or strongly positive reported-negative glandular staining is suspect (HPA tissue IHC; HPA subcellular).
| Membrane topology and compartment | REEP2 has 2 transmembrane segments (UniProt Q9BRK0 topology). HPA reports cytoplasmic neuronal IHC and approved ER localization by ICC-IF (HPA tissue IHC; HPA subcellular); a membrane-protein annotation alone does not specify a cell-surface IHC pattern. |
| Choice of comparison tissue | Caudate and cortical neurons and cerebellar Purkinje cells are High; hippocampal neurons are Medium (HPA tissue IHC). Compare the same reported cell populations, since whole-section darkness can obscure the relevant distribution (general IHC practice). |
| Antibody-specific evidence | HPA031813 is IHC Enhanced; HPA076571 is IHC Supported (HPA antibodies). HPA's overall tissue reliability is Enhanced, but its assessment notes presumed off-target binding; interpret unexpected staining in the context of the antibody used (HPA tissue IHC). |
| Isoforms and epitope interpretation | UniProt lists 2 isoforms (UniProt Q9BRK0). The supplied sources do not map an IHC antibody epitope to either isoform, so an unexpected distribution cannot be assigned to isoform-specific staining from this record alone. |
| IF/ICC Q&A: should the same compartment appear? | Yes: HPA reports approved ER localization by ICC-IF (HPA subcellular), consistent with its cytoplasmic neuronal IHC profile (HPA tissue IHC). The ICC-IF result informs compartment interpretation; it does not establish a paraffin-section IF procedure. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive neuronal populations show no color. | A technical failure is possible when populations reported as High are blank (HPA tissue IHC; general IHC practice). | Check section integrity and detection controls, then compare the antibody dilution and antigen-retrieval steps with the antibody's IHC-P instructions (general IHC practice). |
| Signal is weak in hippocampal neurons but clear in caudate neurons. | HPA reports hippocampal neurons as Medium and caudate neurons as High (HPA tissue IHC). | Score intensity by the relevant cell population and tissue; retain the stronger reported population as the positive comparison (HPA tissue IHC; general IHC practice). |
| Strong nuclear signal dominates the section. | That compartment conflicts with cytoplasmic neuronal IHC and approved ER ICC-IF localization (HPA tissue IHC; HPA subcellular). | Review the negative detection control, antibody concentration, and staining distribution before accepting the signal as REEP2 (general IHC practice). |
| Reported-negative glandular cells stain strongly. | Adrenal, appendix, and duodenal glandular cells are Not detected in HPA tissue IHC; off-target binding or endogenous detection activity are possibilities (HPA tissue IHC; general IHC practice). | Compare with a negative detection control and check the detection system's endogenous-activity blocking step (general IHC practice). |
| Uniform haze obscures neuronal outlines. | Diffuse background prevents assessment of the reported cytoplasmic neuronal pattern (HPA tissue IHC); incomplete blocking, excess antibody, or inadequate washing may contribute (general IHC practice). | Inspect the negative control, then review blocking, antibody dilution, washing, and chromogen development against the chosen IHC workflow (general IHC practice). |
| Two antibodies give different distributions. | HPA assigns different IHC validation statuses to HPA031813 and HPA076571, and notes presumed off-target binding in tissue staining (HPA antibodies; HPA tissue IHC). | Record the antibody used for each section and compare cell type and compartment against HPA's reported pattern before interpreting discordant staining (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing REEP2 staining
A11618 · IHCImmunohistochemistry of REEP2 in rat lung tissue with REEP2 antibody at 5 μg/mL.
A11618 · IFImmunofluorescence of REEP2 in rat lung tissue with REEP2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132563-REEP2/tissue/cerebral+cortex#imid_1438
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132563-REEP2/tissue/cerebral+cortex#imid_8136104
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132563-REEP2/tissue/colon#imid_8136022
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132563-REEP2/tissue/liver#imid_8136031
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA031813; green = REEP2, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA031813; green = REEP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for REEP2 IHC & IF
🧪Run caudate first and expect staining in neuronal cells (HPA: High in caudate neuronal cells); use adrenal gland glandular cells as the negative tissue comparator (HPA: Not detected in adrenal gland glandular cells). Morphologically identified cells outside the expected neuronal staining pattern on the caudate slide should show only background-level signal (HPA: High in caudate neuronal cells; standard IHC control practice).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show REEP2 in HEK293, SH-SY5Y, U2OS, with annotated localisation: Endoplasmic reticulum (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; an irrelevant immunoglobulin matched to the catalog antibody’s host species and antibody type; and REEP2 knockout tissue or peptide competition if a validated blocking peptide is available (standard IHC control practice). Quench endogenous peroxidase for chromogenic detection, and assess neuronal lipofuscin autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the rat lung IHC caption for A11618 also does not state a fixative (selected-SKU tissue-IHC caption). Retrieval dependence is unreported, so optimize antigen retrieval empirically for paraffin sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; IF has an approved ER localisation reference, while brain neuronal lipofuscin can complicate fluorescence scoring (HPA: Endoplasmic reticulum, approved; standard IF practice).
HPA tissue IHC evidence for REEP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced REEP2 IHC Tips
Use compartment, cell-type and control patterns together when optimizing REEP2 staining in paraffin sections (UniProt Q9BRK0; HPA tissue IHC).
How should I optimize retrieval when REEP2 staining is weak in paraffin sections?
Start paraffin-section IHC with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule: cytoplasmic / membrane antigen). Keep section thickness, cooling and detection conditions matched while comparing retrieval runs, so a change in staining can be attributed to retrieval (standard IHC practice). REEP2 has 2 annotated transmembrane segments, but topology alone does not establish which retrieval condition preserves this antibody’s epitope (UniProt Q9BRK0 topology). If staining remains weak, compare a more intensive retrieval condition on adjacent sections and assess tissue damage alongside signal (standard IHC practice). Include a known positive neuronal region and a negative control in the same run (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or inconsistent REEP2 staining?
Target-specific fixation sensitivity for this IHC antibody is unknown from the supplied evidence; the rat-lung caption does not state its fixative (A11618 tissue-IHC caption). Record fixative, fixation duration and processing history for every specimen before comparing staining intensity (standard IHC practice). For formalin-fixed paraffin sections, compare similarly processed positive tissue across runs, because fixation and processing can alter epitope accessibility (standard IHC practice). Avoid interpreting a weak section as REEP2-negative until retrieval, antibody concentration and detection have been checked with matched controls (standard IHC practice). Do not infer a fixation effect from neuronal staining patterns, transmembrane topology or the annotated phosphoserine (HPA tissue IHC; UniProt Q9BRK0).
What cellular pattern should count as plausible REEP2 staining?
Prioritize a cytoplasmic, ER-compatible pattern in neuronal cells when assessing chromogenic IHC (HPA tissue IHC; HPA subcellular). HPA reports high staining in caudate neuronal cells, cerebellar Purkinje cells and cerebral-cortex neuronal cells, providing useful positive comparators (HPA tissue IHC). REEP2 is annotated as a membrane protein with 2 transmembrane segments, and HPA approves ER localisation (UniProt Q9BRK0 topology; HPA subcellular). A sharp nuclear-only pattern or diffuse signal unrelated to cell boundaries should prompt review of controls and detection conditions before scoring (HPA subcellular; standard IHC practice). Compare the pattern with neighboring cells and tissue morphology rather than relying on chromogen intensity alone (standard IHC practice).
Could isoforms or epitope accessibility change my IHC result?
REEP2 has 2 annotated isoforms and transmembrane segments at residues 1–21 and 35–55 (UniProt Q9BRK0). Establish which sequence the catalog antibody recognizes before treating discordant tissue staining as an isoform difference (standard IHC practice). An epitope near a membrane-spanning segment may require different accessibility than one exposed on a soluble side, but the antibody’s epitope is not supplied here (UniProt Q9BRK0 topology; standard IHC practice). UniProt also annotates phosphoserine at residue 150; that annotation alone does not show whether this antibody distinguishes phosphorylation states (UniProt Q9BRK0). Compare staining using matched processing and an independent specificity control before assigning a molecular explanation (standard IHC practice).
How can I use IF to check the REEP2 IHC pattern?
Use IF/ICC as a separate validation experiment and multiplex REEP2 with a neuronal marker when testing the expected positive cell population (HPA tissue IHC; standard IF practice). HPA reports an approved ER location and IF images in HEK293, SH-SY5Y and U2OS cells, which can guide compartment-level comparisons (HPA subcellular). Select spectrally separated fluorophores and inspect unstained tissue for autofluorescence before attributing a channel to REEP2 (standard IF practice). Set permeabilisation according to the antibody epitope’s membrane-facing side; the epitope orientation is unspecified, despite REEP2’s 2 transmembrane segments (UniProt Q9BRK0 topology; standard IF practice). Keep single-stain and secondary-only controls to evaluate bleed-through and background (standard IF practice).
How do I distinguish REEP2 signal from chromogenic background?
First inspect a no-primary control and a known positive section processed with the same detection reagents (standard IHC practice; HPA tissue IHC). Block endogenous peroxidase before peroxidase-based chromogenic detection, and check whether pigment or precipitate persists without primary antibody (standard IHC practice). Titrate the IHC-validated antibody and detection reagents if background rises across unrelated cells or tissue structures (standard IHC practice). The A11618 rat-lung image used 5 µg/mL, but its caption does not establish an optimum for other specimens (A11618 tissue-IHC caption). HPA reports presumed off-target binding that was disregarded, so compartment and cell-type agreement remain important alongside signal strength (HPA tissue IHC).
How should I quantify REEP2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, using neuronal cytoplasmic staining as the main reference pattern (HPA tissue IHC). For cell-level analysis, report the percentage of positive cells and an H-score from intensity categories applied consistently across sections (standard IHC practice). For spatial analysis, report positive-cell density per mm² of viable tissue or a prespecified anatomical region (standard IHC practice). Normalize counts to the number of evaluable cells or measured tissue area, and keep retrieval, imaging and threshold settings matched (standard IHC practice). Record tissue region because HPA reports high neuronal staining in caudate, cerebellum and cerebral cortex, but low staining in lung alveolar cells (HPA tissue IHC).
When is an apparent REEP2-positive IHC result convincing?
A convincing result matches the expected cytoplasmic, ER-compatible compartment and a plausible cell population, particularly neurons in supported brain regions (HPA tissue IHC; HPA subcellular). Treat nuclear-only staining, staining confined to section edges, or color concentrated in necrotic areas as patterns requiring control review (HPA subcellular; standard IHC practice). Exclude endogenous enzyme activity with a no-primary control and appropriate peroxidase blocking before calling chromogen deposition target-specific (standard IHC practice). Interpret rat-lung staining cautiously: the A11618 caption shows staining at 5 µg/mL, while HPA lists lung alveolar cells as low (A11618 tissue-IHC caption; HPA tissue IHC). HPA’s enhanced reliability assessment also notes presumed off-target binding, so morphology alone cannot prove specificity (HPA tissue IHC).
Boster reagents
Best REEP2 / Receptor expression-enhancing protein 2 IHC Antibodies
A11618 has rat lung IHC and IF images (catalog image captions); its listed reactivity covers human, mouse, and rat (catalog: reactivity).
Real IHC data
Anti-REEP2 Antibody
Cat # A11618
A11618 is listed for IHC-P and IF, with human, mouse, and rat reactivity (catalog: applications and reactivity). Its IHC and IF images show rat lung tissue at 5 μg/mL and 20 μg/mL, respectively (catalog image captions).
Which to pick: Choose A11618 for paraffin-section IHC because IHC-P is listed and its own IHC image shows rat lung tissue at 5 μg/mL; the caption does not report the fixative (catalog: applications; IHC image caption). For IF, A11618 has a rat lung tissue image at 20 μg/mL, but ICC validation is not listed (catalog: applications; IF image caption). It is the only listed cross-species choice, with human, mouse, and rat reactivity; its host is rabbit, and clonality is unspecified (catalog: reactivity, host, clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BRK0 (REEP2_HUMAN, Receptor expression-enhancing protein 2).
- Human Protein Atlas. REEP2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. REEP2 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. REEP2 antibody validation summary (2 antibodies).
- M6A-mediated molecular patterns and tumor microenvironment infiltration characterization in nasopharyngeal carcinoma. Cancer biology & therapy 2024 — PMC10978033.
- PubMed PMID:15550249 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:11161817 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for REEP2 (Receptor expression-enhancing protein 2). Expected localisation Neuronal cytoplasm (HPA tissue IHC); membrane protein (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A11618 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Adrenal gland. 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 A11618); 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-REEP2, 5 μg/mL (datasheet A11618); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining was observed (HPA tissue IHC); Isoform / epitope: 2 isoforms ; epitope side affects access; map unknown (UniProt); Regulation: Staining regulation is unreported (UniProt). Evidence: UniProt Q9BRK0, HPA tissue IHC · Enhanced, HPA ICC-IF · endoplasmic reticulum., Datasheet A11618, PMC10978033. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/REEP2/immunohistochemistry-ihc-guide.