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RIMS2 / Regulating synaptic membrane exocytosis protein 2 · IHC design guide
Design Immunohistochemistry for RIMS2
Plan chromogenic RIMS2 IHC in paraffin sections using adrenal zona fasciculata as a high-staining reference (HPA tissue IHC). Assess cytoplasmic adrenal staining (HPA tissue IHC) while accounting for reported off-target binding and splice or transcript discrepancies (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RIMS2 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 | Adrenal cortical cytoplasm (HPA tissue IHC); presynaptic membrane (UniProt) | |
| Staining pattern | Zona fasciculata cells: strong cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+2 more · see all | |
| Negative control | Adipose tissue+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 binding; assess staining specificity (HPA tissue IHC) | |
| Regulation | Fetal brain predominance (UniProt) | |
| Isoform / epitope | 8 isoforms; no transmembrane segment; map the epitope (UniProt) |
Section 1
Recommended RIMS2 IHC & IF Protocols
The catalog antibody’s datasheet-derived IHC-P protocol is paired with 1 published human-tissue RIMS2 IHC protocol (PMC7273530).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A09915); 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-RIMS2, 5 μg/mL (datasheet A09915) |
| 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 | RIMS2-positive staining in cells in zona fasciculata of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in adrenal cortex. 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 rule). The published protocol used Cell Conditioning 2 (PMC7273530).
Section 2
What Is the Expected RIMS2 Staining Pattern?
RIMS2 is associated with synapses and the presynaptic cell membrane, with no transmembrane segment (UniProt Q9UQ26 topology). In tissue IHC, expect cytoplasmic staining in adrenal zona fasciculata cells, with staining also reported in ciliated cell bodies (HPA tissue IHC). HPA rates the tissue profile Enhanced, while noting presumed off-target staining and a splice or transcript discrepancy (HPA tissue IHC).
| Strong cytoplasmic staining in adrenal zona fasciculata cells. | This matches HPA’s reported high staining and adrenal cortex cytoplasmic profile (HPA tissue IHC). Judge the result by both cell identity and compartment. |
| Predominantly nuclear staining, or staining confined to extracellular material. | Neither is a reported RIMS2 location (UniProt Q9UQ26; HPA subcellular). Treat it as suspect; check background and staining controls before scoring it as positive (general IHC practice). |
| Strong staining in an HPA-listed negative cell population. | Possible causes include cross-reactivity or endogenous detection activity (general IHC practice). Compare the specific cell type, since an HPA negative call does not describe every cell in that tissue (HPA tissue IHC). |
| Diffuse chromogen across cells and surrounding tissue, without a distinct cellular pattern. | This limits interpretation even if the slide is dark (general IHC practice). Reassess blocking, washes, detection controls and counterstain before assigning RIMS2 staining. |
| No staining in adrenal zona fasciculata cells. | The expected high-staining reference population is absent (HPA tissue IHC). Check section quality, primary antibody conditions and detection with a known-positive section (general IHC practice). |
💡Expected RIMS2 appearanceA convincing positive shows high cytoplasmic staining in adrenal zona fasciculata cells (HPA tissue IHC); isolated strong nuclear or extracellular signal is inconsistent with the reported locations (UniProt Q9UQ26; HPA subcellular).
| Reference tissue and cell type | HPA reports high staining in adrenal zona fasciculata and nasopharyngeal ciliated cell bodies, medium in bronchial ciliated cell bodies, and low in fallopian tube ciliated cell bodies (HPA tissue IHC). Compare like cells when assessing intensity. |
| Compartment | The adrenal IHC profile is cytoplasmic (HPA tissue IHC). UniProt places RIMS2 at synapses and the presynaptic cell membrane (UniProt Q9UQ26); those annotations do not require every positive paraffin section to show a crisp membrane rim. |
| Antibody evidence | HPA066498 has Enhanced IHC validation; CAB079049 is Supported for IHC (HPA antibodies). HPA also flags presumed off-target staining and a splice or transcript discrepancy, so resolve ambiguous patterns against cell-specific controls (HPA tissue IHC). |
| Isoforms and epitope coverage | UniProt lists 8 RIMS2 isoforms (UniProt Q9UQ26). Epitope coverage for these antibodies is not supplied here; do not treat a negative cell population as proof that every RIMS2 isoform is absent. |
| IF/ICC: what location is reported? | HPA reports approved plasma membrane and cytosol localization in ICC-IF, with images in HeLa, RT-4 and U2OS cells (HPA subcellular). Its ICC-approved antibody is HPA046538; that approval does not establish IHC performance (HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Adrenal reference section has no zona fasciculata signal. | The section misses an HPA high-staining cell population, or the staining run failed (HPA tissue IHC; general IHC practice). | Confirm cell identity and section quality; review the primary antibody conditions and a positive detection control (general IHC practice). |
| A suspected negative control stains strongly. | HPA negative calls are cell-specific; alternatively, nonspecific binding or endogenous detection activity may contribute (HPA tissue IHC; general IHC practice). | Check the named cell population and include an appropriate primary-omission or detection control (general IHC practice). |
| Cerebellar granular layer is negative despite reported cerebellar RIMS2. | HPA reports granular-layer cells as not detected, while UniProt reports protein in Purkinje cells (HPA tissue IHC; UniProt Q9UQ26). | Score the identified cell populations separately; do not use granular-layer negativity to dismiss a Purkinje-cell result. |
| Signal is mainly nuclear. | A nuclear pattern lacks support from the supplied localization records (UniProt Q9UQ26; HPA subcellular). | Review controls and compartment boundaries, then repeat with an IHC-validated antibody if the pattern persists (HPA antibodies; general IHC practice). |
| Chromogen appears broadly diffuse. | Background may reflect nonspecific binding, incomplete washing or detection activity (general IHC practice). | Review blocking, washes and detection controls; score RIMS2 only where cell and compartment remain interpretable (general IHC practice). |
| Two antibodies give different cell patterns. | HPA flags presumed off-target binding and a splice or transcript discrepancy; its IHC antibody validation levels also differ (HPA tissue IHC; HPA antibodies). | Compare staining in the same named cells and reference section, document each antibody separately, and avoid combining discordant scores (general IHC practice). |
Experiment evidence
IHC & IF images showing RIMS2 staining
A09915 · IHCImmunohistochemistry of RIM2 in human adrenal gland tissue with RIM2 antibody at 5 μg/mL.
A09915-1 · IHCImmunohistochemistry of Rim2 in human brain with Rim2 antibody at 5 μg/mL.
A09915-1 · IFImmunofluorescence of Rim2 in Human Brain tissue with Rim2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176406-RIMS2/tissue/cerebral+cortex#imid_19929221
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000176406-RIMS2/tissue/adrenal+gland#imid_250
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176406-RIMS2/tissue/colon#imid_19929155
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176406-RIMS2/tissue/liver#imid_19929156
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA046538; green = RIMS2, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA046538; green = RIMS2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RIMS2 IHC & IF
🧪Run adrenal gland first: cells in the zona fasciculata should stain (HPA: High in cells in zona fasciculata). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the adrenal slide, assess morphologically distinct cells outside the zona fasciculata for background, without assuming they are RIMS2-negative.
Positive control tissue: Adrenal gland (Cells in zona fasciculata, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RIMS2 in HeLa, RT-4, U2OS, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a concentration-matched isotype control appropriate to the primary antibody’s host species and clonality, and a RIMS2 knockout control where available. For chromogenic detection in adrenal tissue, quench endogenous peroxidase and inspect pigment for signal that could be mistaken for staining (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected adrenal IHC caption does not state a fixative (A09915 caption: 5 μg/mL; fixative unreported). Retrieval dependency and whether frozen sections or IF are easier are unreported; compare retrieval conditions on matched sections before scoring (standard IHC practice). Adrenal pigment can complicate chromogenic interpretation, so compare staining with the no-primary slide (standard IHC practice).
HPA tissue IHC evidence for RIMS2
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. Caution, Splice and/or transcript discrepancy exists.). 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 |
|---|---|---|---|---|
| Adrenal gland | Cells in zona fasciculata | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RIMS2 IHC Tips
Troubleshoot RIMS2 staining in paraffin sections by comparing cellular pattern, compartment, and controls with the available IHC evidence.
How should I retrieve RIMS2 in paraffin sections with weak staining?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in buffer, then compare staining with a section processed identically except for primary antibody (general IHC practice). Use adrenal cortex as an anatomical reference: the reported pattern is cytoplasmic, with high staining in zona fasciculata cells (HPA tissue IHC). If signal remains weak, compare a carefully controlled alternative retrieval condition on adjacent sections, keeping antibody concentration and detection constant; no alternative buffer is established for this antibody in the supplied evidence.
Could fixation explain weak or uneven RIMS2 staining?
RIMS2-specific sensitivity to fixation is unknown because no target-specific fixation condition or comparison is supplied. For paraffin IHC, record the fixative, fixation interval, section age, and retrieval run before attributing a weak result to the antigen (general IHC practice). Compare adjacent sections from the same block using the page’s citrate pH 6.0, 95–98 °C, 20 min retrieval setting and identical detection (page retrieval setting; general IHC practice). The adrenal-gland image caption reports antibody at 5 μg/mL but does not state a fixative, so it cannot establish a fixation optimum (A09915 caption).
Should RIMS2 appear on membranes or throughout the cytoplasm?
Interpret the pattern in its cellular context: RIMS2 is associated with synapses and the presynaptic cell membrane (UniProt Q9UQ26 localisation), while the reported adrenal-cortex IHC pattern is cytoplasmic (HPA tissue IHC). Plasma-membrane and cytosol localisation are also approved in the subcellular dataset (HPA subcellular). RIMS2 has no transmembrane segment, so diffuse staining of every cell boundary alone deserves scrutiny (UniProt Q9UQ26 topology; general IHC interpretation). Compare compartment staining in the same tissue with a no-primary control and examine whether zona fasciculata cells show the expected high signal (HPA tissue IHC; general IHC practice).
How can isoforms or epitope masking affect RIMS2 IHC?
RIMS2 has 8 reported isoforms, so check whether the antibody’s documented immunogen is present in the isoform relevant to the sample before treating a negative section as absence of protein (UniProt Q9UQ26 isoforms; general IHC practice). Its Rab-binding, PDZ, and 2 C2 regions provide a domain map for locating a disclosed epitope (UniProt Q9UQ26 domains). The supplied product caption does not identify an epitope, so isoform coverage cannot be assigned to A09915 (A09915 caption). If staining varies with retrieval, compare adjacent sections under controlled conditions and interpret the change as an assay effect until specificity is established (general IHC practice).
How should I plan a follow-up RIMS2 immunofluorescence experiment?
Use immunofluorescence as a separate validation experiment: multiplex RIMS2 with a marker for the cell population being assessed, such as zona fasciculata cells in an adrenal section (HPA tissue IHC; general IF practice). Choose a fluorophore channel after inspecting unstained tissue for autofluorescence, and include single-label controls to check bleed-through (general IF practice). Because RIMS2 has no transmembrane segment and has reported cytosolic and plasma-membrane localisation, test permeabilisation for access to intracellular epitopes while preserving cell outlines (UniProt Q9UQ26 topology; HPA subcellular; general IF practice). The IHC caption’s 5 μg/mL concentration is tissue-IHC evidence, not an established IF dilution (A09915 caption).
What should I change when RIMS2 DAB staining is widespread?
Check the no-primary control first, then examine whether brown signal tracks tissue edges, folds, damaged areas, or the expected cells (general IHC practice). Include a peroxidase block before DAB detection and assess nonspecific binding with an appropriate protein block and controlled primary-antibody titration (general chromogenic IHC practice). The caption documents adrenal-gland IHC at 5 μg/mL, but gives no background comparison or optimal working range (A09915 caption). HPA reports presumed off-target binding that was disregarded, so uniform staining across cell populations warrants particular caution when assigning RIMS2 specificity (HPA tissue IHC reliability description).
How can I quantify RIMS2 staining without overstating specificity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring; zona fasciculata cells have reported high staining, whereas adrenal-cortex expression is described as cytoplasmic (HPA tissue IHC). Score the percentage of positive cells and intensity with an H-score, or measure positive area per mm² when cell segmentation is unreliable (general IHC quantification practice). Normalise to the number of evaluable target cells or sampled tissue area, and keep section thickness, retrieval, exposure, and DAB development consistent across groups (general IHC practice). Report background-subtracted results and the scoring threshold because HPA notes presumed off-target staining and splice or transcript discrepancies (HPA tissue IHC reliability description).
When is a RIMS2-positive cell convincing rather than artefactual?
A convincing adrenal result aligns with cytoplasmic staining in zona fasciculata cells, which are reported as high, while accounting for the documented adrenal-cortex pattern (HPA tissue IHC). Cytosolic or plasma-membrane signal is biologically plausible, and synaptic localisation may matter in neural tissue (HPA subcellular; UniProt Q9UQ26 localisation). Investigate signal confined to section edges, folds, necrotic areas, or no-primary controls, as well as diffuse nuclear-only or endogenous-peroxidase-like DAB deposits (general IHC practice; HPA subcellular). Treat isolated unexpected positives cautiously because HPA reports presumed off-target binding and a splice or transcript discrepancy; corroborate with an independent specificity control where possible (HPA tissue IHC reliability description; general IHC practice).
Boster reagents
Best RIMS2 / Regulating synaptic membrane exocytosis protein 2 IHC Antibodies
Two anti-RIMS2 antibodies have human tissue IHC images: adrenal gland for A09915 and brain for A09915-1; A09915-1 also has a human brain IF image (catalog image captions).
Real IHC data
Anti-RIM2 RIMS2 Antibody
Cat # A09915
Real IHC data
Anti-Rim2 RIMS2 Antibody
Cat # A09915-1
A09915 has a human adrenal gland IHC image at 5 μg/mL (A09915 IHC image caption). A09915-1 has human brain IHC and IF images at 5 μg/mL and 20 μg/mL, respectively (A09915-1 image captions).
Which to pick: For human paraffin section IHC, either SKU lists IHC-P; choose A09915 for the adrenal gland image or A09915-1 for the brain image (catalog applications; each SKU’s IHC image caption). Choose A09915-1 for tissue IF or work involving mouse or rat because it lists IF and those species; ICC is not listed (A09915-1 applications and reactivity). A09915 is described as polyclonal, while A09915-1 has no clone designation; neither IHC caption reports a fixative (A09915 dilution text; A09915-1 clone field; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UQ26 (RIMS2_HUMAN, Regulating synaptic membrane exocytosis protein 2).
- Human Protein Atlas. RIMS2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RIMS2 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. RIMS2 antibody validation summary (3 antibodies).
- Loss of Function of RIMS2 Causes a Syndromic Congenital Cone-Rod Synaptic Disease with Neurodevelopmental and Pancreatic Involvement. American journal of human genetics 2020 — PMC7273530.
- PubMed PMID:9872452 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RIMS2 (Regulating synaptic membrane exocytosis protein 2). Expected localisation Adrenal cortical cytoplasm (HPA tissue IHC); presynaptic membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A09915 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Cells in zona fasciculata); negative Adipose tissue. 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 A09915); 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-RIMS2, 5 μg/mL (datasheet A09915); 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 binding; assess staining specificity (HPA tissue IHC); Isoform / epitope: 8 isoforms; no transmembrane segment; map the epitope (UniProt); Regulation: Fetal brain predominance (UniProt). Evidence: UniProt Q9UQ26, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and cytosol., Datasheet A09915, PMC7273530. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RIMS2/immunohistochemistry-ihc-guide.