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ERLIN2 / Erlin-2 · IHC design guide
Design Immunohistochemistry for ERLIN2
Plan ERLIN2 paraffin-section IHC around cytoplasmic tissue staining (HPA tissue IHC) and ER membrane localisation (UniProt). Use breast glandular cells as a high-staining reference and cardiomyocytes as a not-detected reference when evaluating staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ERLIN2 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Ubiquitous cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M07042-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Heart muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image M07042-1) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 3 isoforms; antibody epitope is unspecified (UniProt) |
Section 1
Recommended ERLIN2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published ERLIN2 workflows for lung adenocarcinoma and breast tissue microarrays (PMC7793841; PMC10818095).
| Sample | Paraffin-embedded human ovarian serous carcinoma tissue; fixative not specified (datasheet M07042-1) |
| Fixation | Image fixative and duration unreported (datasheet M07042-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M07042-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M07042-1) |
| Primary antibody | Mouse monoclonal (clone 3H9A2) anti-ERLIN2, 2 μg/ml (datasheet M07042-1) |
| Primary incubation | Overnight at 4 °C (datasheet M07042-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M07042-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ERLIN2-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet M07042-1); compare citrate at pH 6.0 if staining needs optimization (PMC10818095).
Section 2
What Is the Expected ERLIN2 Staining Pattern?
ERLIN2 is an endoplasmic reticulum (ER) membrane protein with residues 25–339 facing the ER lumen (UniProt O94905 topology). In paraffin sections, expect cytoplasmic staining across many cell types: HPA describes ubiquitous cytoplasmic expression, with high staining in breast glandular cells and bronchial respiratory epithelial cells (HPA tissue IHC). HPA rates its tissue IHC profile Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in breast glandular cells or bronchial respiratory epithelial cells. | This matches the reported high staining in both cell types (HPA tissue IHC). An ER-associated distribution is biologically consistent with ERLIN2's membrane location (UniProt O94905); chromogenic IHC alone cannot prove that each stained structure is ER. |
| Predominantly nuclear or sharply plasma-membrane staining, with little cytoplasmic signal. | That compartment conflicts with ERLIN2's ER membrane location (UniProt O94905) and HPA's cytoplasmic tissue profile (HPA tissue IHC). Check the negative control and antibody specificity before scoring it as ERLIN2. |
| Strong cardiomyocyte staining, especially when expected positive cells also stain. | HPA reports ERLIN2 as not detected in cardiomyocytes (HPA tissue IHC). Consider cross-reactivity or endogenous chromogen activity; compare matched controls and do not treat one unexpected cell type as proof of a new expression pattern. |
| Diffuse stain covers connective tissue, extracellular space, and cells alike. | A broad haze lacks the cell-associated cytoplasmic pattern reported by HPA (HPA tissue IHC). Assess background with a primary-antibody omission control, then review blocking, washes, and detection conditions as general IHC checks. |
| No visible stain in breast glandular cells or bronchial respiratory epithelial cells. | Both are reported high-staining cell populations (HPA tissue IHC), so their absence raises a technical concern. Confirm tissue identity and control performance, then review retrieval, antibody dilution, and detection; a single negative section cannot establish ERLIN2 absence. |
💡Expected ERLIN2 appearanceCall a section positive when cellular cytoplasmic staining is clear in an expected high-staining population, such as breast glandular cells (HPA tissue IHC), and is compatible with ER localization (UniProt O94905); isolated nuclear signal or uniform extracellular haze warrants investigation as possible artifact.
| Which compartment should guide scoring? | Score cell-associated cytoplasmic signal: ERLIN2 resides in the ER membrane (UniProt O94905), while HPA describes ubiquitous cytoplasmic tissue staining (HPA tissue IHC). Routine chromogenic IHC may not resolve the ER network. |
| How should tissue choice affect interpretation? | Breast glandular and bronchial respiratory epithelial cells are reported High; cardiomyocytes are Not detected (HPA tissue IHC). Use the former as expected-positive examples and interpret the latter at cell-type level, not as a whole-heart negative. |
| What does the antibody validation establish? | HPA lists IHC as Supported for HPA002025 and CAB014894, and rates the tissue profile Supported with medium staining–RNA consistency (HPA tissue IHC). These ratings support use of the reported pattern but do not verify every signal in a new section. |
| Does sequence information predict an IHC failure? | ERLIN2 has one transmembrane segment at residues 4–24, three isoforms, and a glycosylation site at residue 106 (UniProt O94905). The supplied sources give no antibody epitope or target-specific fixation-sensitivity result, so these features do not justify a retrieval or staining prediction. |
| IF/ICC: what localization is expected? | Mainly ER localization is reported and supported in HPA ICC-IF (HPA subcellular). This answers the localization question only; IF/ICC conditions belong in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected-positive tissue has no signal. | A failed staining step is possible when high-staining cells are absent from the result (HPA tissue IHC). | Check section identity and assay controls. As general IHC practice, review retrieval, primary dilution, detection reagents, and counterstain before interpreting a biological negative. |
| Signal is mostly nuclear. | Nuclear dominance does not fit the ER membrane assignment (UniProt O94905) or HPA's cytoplasmic profile (HPA tissue IHC). | Compare the expected-positive cells and omission control; review antibody specificity and scoring before accepting the nuclear signal. |
| Cardiomyocytes appear strongly positive. | HPA reports cardiomyocytes as Not detected (HPA tissue IHC); unexpected signal may reflect nonspecific binding or endogenous detection activity. | Inspect cell identity and matched controls. If the control also colors cardiomyocytes, troubleshoot the detection system before assigning ERLIN2 positivity. |
| The whole section has diffuse brown background. | Uniform staining obscures the cell-associated cytoplasmic pattern reported for ERLIN2 (HPA tissue IHC). | Use an omission control, then review blocking, wash stringency, and chromogen development as general chromogenic IHC checks. |
| Only low-staining tissue looks negative. | HPA lists squamous epithelial cells of oral mucosa and vagina as Low, while selected glandular and respiratory epithelial cells are High (HPA tissue IHC). | Run an expected high-staining cell population alongside it; score the named cell type rather than declaring a whole tissue negative. |
| Staining is present but its intracellular pattern is unclear. | The ER membrane assignment (UniProt O94905) and cytoplasmic IHC profile (HPA tissue IHC) set the expectation, but chromogenic resolution may limit compartment assignment. | Record the observable cytoplasmic distribution and control results. For finer localization, consult the separate IF/ICC guide; HPA reports supported ER localization by ICC-IF (HPA subcellular). |
Experiment evidence
IHC & IF images showing ERLIN2 staining
M07042-1 · IHCIHC analysis of ERLIN2 using anti-ERLIN2 antibody (M07042-1). ERLIN2 was detected in a paraffin-embedded section of human ovarian serous car
M07042-1 · IHCIHC analysis of ERLIN2 using anti-ERLIN2 antibody (M07042-1). ERLIN2 was detected in a paraffin-embedded section of human placenta tissue. H
M07042-1 · IFIF analysis of ERLIN2 using anti-ERLIN2 antibody (M07042-1). ERLIN2 was detected in an immunocytochemical section of T47D cells. Enzyme anti
A07042-2 · IFIF analysis of Erlin-2/ERLIN2 using anti-Erlin-2/ERLIN2 antibody (A07042-2). Erlin-2/ERLIN2 was detected in an immunocytochemical section of
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000147475-ERLIN2/tissue/cerebral+cortex#imid_941600
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000147475-ERLIN2/tissue/colon#imid_941518
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000147475-ERLIN2/tissue/liver#imid_941519
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000147475-ERLIN2/tissue/kidney#imid_941536
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002025; green = ERLIN2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA002025; green = ERLIN2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ERLIN2 IHC & IF
🧪Run breast first: glandular cells should stain (HPA: High in breast glandular cells). Use heart muscle as the negative tissue, focusing on cardiomyocytes (HPA: Not detected in cardiomyocytes); on the breast slide, treat any non-glandular cells as an internal negative reference only if they show background-level staining, since their ERLIN2 status is not specified by the supplied HPA row.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ERLIN2 in A-431, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a mouse IgG isotype control matched to the catalog primary (selected-SKU caption: mouse anti-ERLIN2), plus ERLIN2-knockout material as a biological negative. Block endogenous peroxidase and check for background DAB signal in the breast section because the illustrated detection uses HRP and DAB (selected-SKU caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not state the fixative. The illustrated paraffin-section procedure used heat retrieval in EDTA at pH 8.0, but retrieval dependence has not been established for the proposed breast section (selected-SKU caption: EDTA heat retrieval). The supplied evidence does not establish whether frozen sections or IF are easier, or identify a breast-specific artefact; HPA does support an ER localization reference for IF/ICC (HPA: endoplasmic reticulum, supported).
HPA tissue IHC evidence for ERLIN2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ERLIN2 IHC Tips
Troubleshoot ERLIN2 chromogenic IHC by checking retrieval, processing, cellular localisation and controls before comparing staining across sections.
What retrieval should I try when ERLIN2 staining is weak in paraffin sections?
Begin heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet M07042-1). The selected image pairs this retrieval with 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption M07042-1). If signal remains weak, compare heating times on adjacent sections while keeping antibody concentration and DAB development constant (standard IHC practice). Review tissue morphology and no-primary background after each condition, since excessive heating can damage sections or raise nonspecific staining (standard IHC practice). Select the condition that preserves morphology and resolves ER-associated cytoplasmic signal (UniProt O94905 localisation; HPA tissue IHC).
Could fixation explain inconsistent ERLIN2 staining between paraffin blocks?
Record the fixative and fixation duration for each block before comparing ERLIN2 intensity across cases (standard IHC practice). The selected paraffin-section caption reports neither the fixative nor fixation time, so ERLIN2-specific fixation sensitivity remains unknown (caption M07042-1). Its ER membrane topology, glycosylation site at residue 106 and tissue staining pattern do not establish a preferred fixative (UniProt O94905; HPA tissue IHC). Compare sections with documented processing histories using the same EDTA pH 8.0 retrieval and 2 μg/ml primary concentration (datasheet M07042-1; caption M07042-1). Evaluate differences alongside morphology and no-primary background before attributing them to expression (standard IHC practice).
Which staining pattern is plausible for ERLIN2 in tissue sections?
Expect predominantly cytoplasmic staining compatible with an ER network, rather than a discrete nuclear signal (UniProt O94905 localisation; HPA tissue IHC). ERLIN2 has a membrane anchor at residues 4–24, with residues 25–339 facing the ER lumen (UniProt O94905 topology). HPA describes ubiquitous cytoplasmic tissue staining and reports high signal in breast glandular cells and bronchial respiratory epithelial cells (HPA tissue IHC). Compare staining within the same cell type and confirm that counterstain and DAB intensity still allow intracellular patterns to be resolved (standard IHC practice). Treat a sharply nuclear-only or extracellular pattern as a reason to recheck background and antibody specificity (UniProt O94905 localisation; standard IHC practice).
How should unknown epitope coverage affect interpretation of ERLIN2 IHC?
ERLIN2 has 3 annotated isoforms, but the supplied antibody caption does not identify the recognised epitope or establish isoform coverage (UniProt O94905 isoforms; caption M07042-1). Its membrane anchor spans residues 4–24, while residues 25–339 are lumenal (UniProt O94905 topology). A glycosylation site is annotated at residue 106; that annotation alone does not show whether this antibody is affected by glycosylation (UniProt O94905 processing). Obtain the immunogen sequence or epitope mapping before assigning an isoform-specific explanation to weak staining (standard IHC practice). Until then, report the result as ERLIN2 immunoreactivity under the stated retrieval and antibody conditions (datasheet M07042-1; standard IHC practice).
How can IF help assess an ambiguous ERLIN2 IHC pattern?
Use IF as a localisation cross-check when chromogenic staining obscures the intracellular pattern; HPA supports ER localisation in ICC/IF images (HPA subcellular). Multiplex ERLIN2 with a marker for the expected cell type, such as respiratory epithelium when evaluating bronchus, and compare signals within identified cells (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and include single-stain and no-primary controls for spectral bleed-through and background (standard IF practice). Match permeabilisation to the antibody epitope: residues 25–339 face the ER lumen, but this antibody’s epitope is unreported (UniProt O94905 topology; caption M07042-1). Optimise IF fixation separately because the paraffin IHC caption provides no IF fixation evidence (caption M07042-1).
How do I reduce diffuse DAB staining without losing ERLIN2 signal?
Run a no-primary section through the same secondary antibody and DAB steps to locate detection-system background (standard IHC practice). Block endogenous peroxidase before HRP detection, then adjust protein blocking, washes or primary concentration one variable at a time (standard IHC practice). The selected image used 10% goat serum, 2 μg/ml mouse primary overnight at 4°C, and an HRP-based DAB readout (caption M07042-1). Because HPA describes broadly distributed cytoplasmic expression, widespread staining alone cannot establish that a signal is nonspecific (HPA tissue IHC). Prefer conditions that preserve intracellular contrast while clearing no-primary staining and tissue-edge deposits (UniProt O94905 localisation; standard IHC practice).
What is a defensible way to score ERLIN2 chromogenic IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then exclude folds, necrosis and poorly preserved areas (standard IHC practice). Record the percentage of positive viable cells and staining intensity, or calculate an H-score from intensity categories and their cell percentages (standard IHC practice). Normalise counts to the number of viable cells in that population; for area-based analysis, report positive cells per mm² of evaluable tissue (standard IHC practice). Keep retrieval, 2 μg/ml primary concentration, DAB development and image settings consistent across compared sections (datasheet M07042-1; caption M07042-1; standard IHC practice). Report cell types separately because HPA tissue staining varies by cell population (HPA tissue IHC).
When should apparent ERLIN2 positivity be treated cautiously?
Favor intracellular, predominantly cytoplasmic staining in intact cells, consistent with ERLIN2’s ER membrane localisation and HPA tissue profile (UniProt O94905 localisation; HPA tissue IHC). Compare the suspected positive population with a no-primary control and with neighbouring cells in the same section (standard IHC practice). Nuclear-only deposits, tissue-edge accentuation and staining confined to necrotic material warrant review for artefact (UniProt O94905 localisation; standard IHC practice). Check endogenous peroxidase blocking when DAB appears in the no-primary control, and assess morphology before assigning a cell type (standard IHC practice). HPA rates tissue staining as supported with medium RNA–protein consistency, so describe discordant cases cautiously (HPA tissue IHC).
Boster reagents
Best ERLIN2 / Erlin-2 IHC Antibodies
ERLIN2 antibody cards show human paraffin-section IHC and human cell IF images (catalog captions); one IF/ICC antibody also lists mouse and rat reactivity (catalog: A07042-2).
Real IHC data
Anti-Erlin-2/ERLIN2 Antibody ® (monoclonal, 3H9A2)
Cat # M07042-1
Real IF data
Anti-Erlin-2/ERLIN2 Antibody ®
Cat # A07042-2
M07042-1 shows IHC staining in human paraffin-embedded ovarian serous carcinoma, placenta, squamous cell lung carcinoma and thyroiditis sections (M07042-1 IHC captions). A07042-2 shows IF staining in human T-47D cells (A07042-2 IF caption).
Which to pick: Choose M07042-1 for human paraffin-section IHC: its own captions show staining after EDTA pH 8.0 retrieval at 2 μg/ml (M07042-1 IHC captions); the fixative is unreported (M07042-1 IHC captions). Choose A07042-2 for human IF/ICC: its own caption shows T-47D cell staining at 5 μg/ml (A07042-2 IF caption). For mouse or rat work, A07042-2 lists both species as reactive, but its pictured IF example is human only (A07042-2 catalog reactivity and IF caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O94905 (ERLN2_HUMAN, Erlin-2).
- Human Protein Atlas. ERLIN2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. ERLIN2 subcellular location (ICC-IF): Mainly localized to the endoplasmic reticulum..
- Human Protein Atlas. ERLIN2 antibody validation summary (2 antibodies).
- Endoplasmic reticulum stress-related genes as prognostic and immunogenic biomarkers in prostate cancer. European journal of medical research 2024 — PMC11031923.
- ERLIN2 promotes breast cancer cell survival by modulating endoplasmic reticulum stress pathways. BMC cancer 2012 — PMC3732090.
- Molecular and Immune Characteristics for Lung Adenocarcinoma Patients With ERLIN2 Overexpression. Frontiers in immunology 2020 — PMC7793841.
- KCNN1 promotes proliferation and metastasis of breast cancer via ERLIN2-mediated stabilization and K63-dependent ubiquitination of Cyclin B1. Carcinogenesis 2023 — PMC10818095.
- PubMed PMID:10449903 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ERLIN2 (Erlin-2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M07042-1). Validated IHC antibody: M07042-1 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian serous carcinoma tissue; fixative not specified (datasheet M07042-1); 2. Fixation: Image fixative and duration unreported (datasheet M07042-1); 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: EDTA pH 8.0 (datasheet M07042-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M07042-1); 8. Primary antibody: Mouse monoclonal (clone 3H9A2) anti-ERLIN2, 2 μg/ml (datasheet M07042-1); 9. Primary incubation: Overnight at 4 °C (datasheet M07042-1). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image M07042-1); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; antibody epitope is unspecified (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt O94905, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the endoplasmic reticulum., Datasheet M07042-1, PMC7793841, PMC10818095. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ERLIN2/immunohistochemistry-ihc-guide.