This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ELOB / Elongin-B · IHC design guide
Design Immunohistochemistry for ELOB
Plan ELOB paraffin IHC with the catalog antibody at 2–5 μg/ml (datasheet A31718-2). Use breast glandular cells as a high-staining reference and assess nuclear and cytoplasmic signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ELOB 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 | Nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Nuclear and cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A31718-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Cerebellum |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has medium RNA concordance (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported (UniProt) |
Section 1
Recommended ELOB IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A31718-2) with the published human breast tissue array protocol (PMC11039524).
| Sample | Paraffin-embedded mouse pancreas tissue; fixative not specified (datasheet A31718-2) |
| Fixation | Image fixative and duration unreported (datasheet A31718-2); 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 A31718-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A31718-2) |
| Primary antibody | Rabbit anti-ELOB, 2-5 μg/ml (datasheet A31718-2) |
| Primary incubation | Overnight at 4 °C (datasheet A31718-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A31718-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ELOB-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A31718-2); the published tissue array protocol does not specify retrieval (PMC11039524).
Section 2
What Is the Expected ELOB Staining Pattern?
ELOB should appear in nuclei, with cytoplasmic staining also reported across tissues (UniProt Q15370: nucleus; HPA: nuclear and cytoplasmic expression in all tissues). Strong staining is reported in glandular cells of several organs, endometrial stromal cells, and bone marrow hematopoietic cells (HPA: High). ELOB has no transmembrane segment (UniProt Q15370 topology). HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA data and external verification pending (HPA: reliability).
| Nuclear staining, with or without cytoplasmic staining, in expected cells. | This fits the reported distribution (UniProt Q15370: nucleus; HPA: nuclear and cytoplasmic expression). Compare cells within the same section before scoring intensity: HPA reports High staining in breast glandular cells and endometrial stromal cells, among others (HPA: tissue IHC). |
| Predominantly membrane-edge or extracellular deposits, without a convincing nuclear signal. | This is outside the reported nuclear and cytoplasmic pattern (UniProt Q15370: nucleus; HPA: tissue IHC). Because ELOB has no transmembrane segment or signal peptide, investigate staining artefact or nonspecific detection before calling these deposits ELOB (UniProt Q15370 topology and processing; general IHC practice). |
| Strong color in a cell population expected to be unstained or substantially weaker. | Check cell identity and the relevant HPA entry before interpreting it: cerebellar granular-layer cells are Not detected, whereas colon endothelial cells are Low (HPA: tissue IHC). Unexpected staining can reflect cross-reactivity or endogenous detection activity (general IHC practice); one discordant cell type cannot establish either cause. |
| Diffuse color across tissue, including spaces or structures without clear cellular boundaries. | A diffuse deposit cannot reliably establish ELOB localization (general IHC practice). Examine a no-primary control and the counterstain to distinguish cellular signal from background; compare with the reported nuclear and cytoplasmic distribution only after background is controlled (HPA: tissue IHC; general IHC practice). |
| No discernible signal in a tissue and cell type reported as High. | For example, absent staining in breast glandular cells or bone marrow hematopoietic cells conflicts with the reported pattern (HPA: High). First check that the expected cells are present, then review the control slide and detection workflow (general IHC practice). A blank slide alone does not demonstrate ELOB absence. |
💡Expected ELOB appearanceCall ELOB positive when expected cells show clear nuclear staining, with possible cytoplasmic signal and stronger staining in HPA High populations; isolated membrane-edge deposits or diffuse color are suspect (UniProt Q15370: nucleus; HPA: tissue IHC; general IHC practice).
| Tissue and cell selection | Use an HPA High population, such as adrenal glandular cells or endometrial stromal cells, to assess whether the assay can show a positive result (HPA: tissue IHC). Cerebellar granular-layer cells are reported Not detected, but surrounding tissue needs its own interpretation (HPA: tissue IHC). |
| Compartment and topology | UniProt lists ELOB in the nucleus and reports no transmembrane segment or signal peptide (UniProt Q15370: subcellular location, topology and processing). HPA tissue IHC also reports cytoplasmic staining, so cytoplasmic color alone should be judged against cell identity and controls rather than rejected automatically (HPA: tissue IHC; general IHC practice). |
| Confidence in the reference pattern | HPA calls the tissue IHC antibody Approved, describing medium consistency with RNA expression and pending external verification (HPA: reliability). Its RNA specificity is Low tissue specificity (HPA: tissue IHC); a reported Not detected cell population should therefore be used as a local comparison, not as proof that an entire organ lacks ELOB. |
| Isoforms and modifications | UniProt lists two isoforms and several modified residues, including phosphorylated residues at 84, 108 and 111 (UniProt Q15370: isoforms and modified residues). The supplied evidence gives no antibody epitope, isoform coverage or modification-sensitive staining result; interpret any proposed distinction only after obtaining assay-specific evidence. |
| IF/ICC: what can be inferred? | UniProt supports nuclear localization, and HPA tissue IHC reports nuclear and cytoplasmic expression (UniProt Q15370: subcellular location; HPA: tissue IHC). HPA supplies no ICC-IF images or main subcellular location for ELOB (HPA: subcellular record), so this IHC pattern is a starting expectation rather than IF/ICC validation. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected High population is blank. | The chosen field may lack the expected cells, or the IHC detection workflow may have failed (HPA: High; general IHC practice). | Confirm cell identity on the counterstain; run a known-positive section alongside the sample and check primary-antibody application and detection reagents (general IHC practice). |
| The whole section has diffuse brown background. | Nonspecific binding or endogenous detection activity may obscure cellular staining (general IHC practice). | Compare a no-primary control; review blocking, washing and endogenous-enzyme blocking appropriate to the chromogenic system, then reassess nuclear and cytoplasmic signal (general IHC practice; HPA: tissue IHC). |
| Color appears mainly at cell borders or outside cells. | That pattern is discordant with the reported compartments and topology (UniProt Q15370: nucleus, no transmembrane segment; HPA: tissue IHC). | Check the no-primary control and section morphology; score ELOB only where a credible cellular signal remains (general IHC practice). |
| Only cytoplasm stains in an expected positive population. | HPA allows cytoplasmic staining, but the absent nuclear component differs from UniProt's listed location (HPA: tissue IHC; UniProt Q15370: nucleus). | Check nuclear preservation and background against the counterstain and controls; describe the observed compartment rather than assigning nuclear positivity (general IHC practice). |
| Cerebellar granular-layer cells appear strongly positive. | This differs from HPA's Not detected report for those cells; cell identification, cross-reactivity or endogenous activity may explain the discrepancy (HPA: tissue IHC; general IHC practice). | Verify the layer on the counterstain, inspect a no-primary control, and compare staining with an HPA High population processed in the same run (HPA: tissue IHC; general IHC practice). |
| A weak population seems indistinguishable from a High one. | Background or detection settings may compress the apparent difference (general IHC practice); HPA reports Low colon endothelial cells and High breast glandular cells (HPA: tissue IHC). | Score comparable cell types and compartments separately, with controls and matched detection conditions; report the observed intensity rather than forcing it into HPA's categories (general IHC practice). |
Experiment evidence
IHC & IF images showing ELOB staining
A31718-2 · IHCIHC analysis of TCEB2/Elongin-B/ELOB using anti-TCEB2/Elongin-B/ELOB antibody (A31718-2). TCEB2/Elongin-B/ELOB was detected in a paraffin-em
A31718-2 · IHCIHC analysis of TCEB2/Elongin-B/ELOB using anti-TCEB2/Elongin-B/ELOB antibody (A31718-2). TCEB2/Elongin-B/ELOB was detected in a paraffin-em
A31718-2 · IFIF analysis of TCEB2/Elongin-B/ELOB using anti-TCEB2/Elongin-B/ELOB antibody (A31718-2). TCEB2/Elongin-B/ELOB was detected in an immunocytoc
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000103363-ELOB/tissue/cerebral+cortex#imid_20626627
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000103363-ELOB/tissue/adrenal+gland#imid_20626619
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000103363-ELOB/tissue/colon#imid_20626553
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000103363-ELOB/tissue/liver#imid_20626562
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ELOB IHC & IF
🧪Run breast first: glandular cells should show strong ELOB staining (HPA: High in breast glandular cells). Use cerebellum as the negative tissue, focusing on granular-layer cells (HPA: Not detected in cerebellar granular-layer cells); within the breast section, cells without specific nuclear signal should show only background staining, but should not be assumed ELOB-negative without validation (UniProt Q15370: nucleus).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Cerebellum (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ELOB; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and matched nonimmune rabbit IgG controls (selected-SKU caption: rabbit primary antibody), plus ELOB-knockout tissue as a biological specificity control. Block endogenous peroxidase for chromogenic detection and assess endogenous biotin if using the caption’s biotinylated secondary and streptavidin–biotin detection (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used, but whether ELOB staining depends on that retrieval is unreported (selected-SKU caption: heat-mediated EDTA retrieval). No matched frozen-section or IF protocol is supplied to establish an easier route, and no breast-specific artefact is documented in the supplied evidence (HPA subcellular: no ICC-IF image cell lines; selected-SKU caption: mouse pancreas IHC).
HPA tissue IHC evidence for ELOB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ELOB IHC Tips
Troubleshoot ELOB staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting differences between samples.
Which retrieval conditions should I try first for weak ELOB staining?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for this catalog antibody (datasheet A31718-2). The demonstrated mouse pancreas workflow then used 2 μg/ml primary antibody overnight at 4°C, so keep those conditions consistent while assessing retrieval (caption A31718-2). If staining remains weak, compare retrieval duration on adjacent sections while holding detection and imaging conditions fixed (standard IHC practice). Include a section processed without primary antibody and a previously staining section to distinguish weak antigen exposure from detection failure or nonspecific signal (standard IHC practice).
How should I investigate inconsistent ELOB staining after fixation?
Record the fixative, fixation duration and processing history for each specimen before comparing staining, because these variables can change antigen accessibility in paraffin sections (standard IHC practice). Target-specific fixation sensitivity for ELOB is unknown from the supplied evidence; the mouse pancreas caption describes a paraffin section but does not state its fixative (caption A31718-2). Compare similarly processed sections with the same EDTA pH 8.0 retrieval and detection conditions to isolate fixation or processing differences (datasheet A31718-2; standard IHC practice). Do not assign a staining difference to a particular fixative without a controlled comparison using this antibody (standard IHC practice).
Should ELOB staining be nuclear, cytoplasmic, or both?
Inspect nuclei carefully because ELOB is annotated as nuclear, while tissue IHC reports nuclear and cytoplasmic expression across tissues (UniProt Q15370 localisation; HPA tissue IHC). Score nuclear and cytoplasmic staining separately instead of treating either compartment alone as proof of antibody specificity (standard IHC practice). ELOB has no transmembrane segment, so a crisp cell-surface pattern would need independent validation before interpretation as ELOB (UniProt Q15370 topology; standard IHC practice). Compare compartment patterns in adjacent intact cells and a no-primary control, using the same counterstain and microscope settings throughout (standard IHC practice).
Can this antibody distinguish ELOB isoforms or modified epitopes in tissue?
Do not interpret staining as isoform-specific without an epitope map or direct validation: ELOB has 2 annotated isoforms, but the supplied caption does not identify the antibody epitope (UniProt Q15370 isoforms; caption A31718-2). Its ubiquitin-like region spans residues 1–66, and annotated modifications include phosphorylation at residues 84, 108 and 111 (UniProt Q15370 domains and modified residues). Those annotations alone do not establish which forms this antibody detects in paraffin sections (standard IHC interpretation). If isoform or modification status matters, pair compartment-specific IHC scoring with a separately validated, form-specific assay (standard IHC practice).
How can I check an ELOB pattern by multiplex immunofluorescence?
Treat IF as a separate assay and validate its fixation, permeabilisation and antibody conditions directly; the supplied tissue demonstration is chromogenic IHC on a paraffin section (caption A31718-2; standard IF practice). Multiplex ELOB with a marker for the expected cell type, such as a glandular-cell marker when examining a glandular compartment, and inspect nuclear overlap with a DNA counterstain (HPA tissue IHC; UniProt Q15370 localisation; standard IF practice). Choose a spectrally separated, preferably far-red fluorophore after checking tissue autofluorescence and single-stain controls (standard IF practice). Permeabilise sufficiently for access to the nuclear epitope, then compare with a no-primary control; ELOB has no transmembrane segment (UniProt Q15370 topology; standard IF practice).
What should I change when diffuse brown staining obscures ELOB?
First compare the section with a no-primary control to identify signal arising from detection reagents or tissue rather than the primary antibody (standard IHC practice). The demonstrated workflow used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin–biotin detection and DAB, so assess blocking and endogenous biotin in that workflow (caption A31718-2; standard IHC practice). Include a peroxidase block before DAB development and inspect whether pigment or precipitate remains in the control (standard chromogenic IHC practice). If primary-dependent background persists, titrate the primary around the demonstrated 2 μg/ml condition while preserving a positive reference section (caption A31718-2; standard IHC practice).
How should I score ELOB across sections with different cellularity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and count intact cells within matched regions before scoring, because cellularity and sampled area can otherwise change apparent signal (standard IHC practice). Report nuclear and cytoplasmic results separately as percent positive cells and an intensity-weighted H-score from 0–300, with a fixed scoring threshold (UniProt Q15370 localisation; HPA tissue IHC; standard IHC practice). If counting positive cells per area, report density per mm² and normalise comparisons to the relevant viable-cell count or defined tissue area (standard IHC practice). Keep retrieval, chromogen development, counterstain and image acquisition consistent across sections, and include the same reference section in each staining run (standard IHC practice).
How do I distinguish genuine ELOB staining from artefact?
Look for reproducible cellular staining in intact tissue, including a nuclear component consistent with ELOB annotation, while recognising that tissue IHC also reports cytoplasmic signal (UniProt Q15370 localisation; HPA tissue IHC). Compare the suspected positive compartment with its cell identity: HPA reports high staining in several glandular-cell populations, but granular-layer cells of the cerebellum were not detected (HPA tissue IHC). Exclude edge-concentrated staining, necrotic regions and signal present without primary antibody before assigning a positive score (standard IHC practice). With DAB detection, check endogenous peroxidase and streptavidin–biotin background using appropriate controls, especially when staining does not follow cell boundaries (caption A31718-2; standard chromogenic IHC practice).
Boster reagents
Best ELOB / Elongin-B IHC Antibodies
The IHC-validated anti-ELOB antibody has paraffin-section images from mouse and rat pancreas and human breast carcinoma and stomach (A31718-2 IHC captions), plus IF data from MCF-7 cells (A31718-2 IF caption).
Real IHC data
Anti-TCEB2/Elongin-B/ELOB Antibody ®
Cat # A31718-2
A31718-2 is listed for IHC in human, mouse and rat, with paraffin-section images from mouse and rat pancreas and human breast carcinoma and stomach (A31718-2 catalog and IHC captions). A31718-2 is also listed for IF/ICC, with an IF image from MCF-7 cells (A31718-2 catalog and IF caption).
Which to pick: For tissue IHC, choose A31718-2: its own captions show paraffin sections stained at 2 μg/ml after EDTA retrieval at pH 8.0; the fixative is unreported (A31718-2 IHC captions). For IF/ICC, choose A31718-2: both applications are listed, and its MCF-7 IF caption uses 5 μg/ml (A31718-2 catalog and IF caption). For cross-species IHC, A31718-2 lists human, mouse and rat reactivity and has IHC images from all three species; the catalog identifies a rabbit host but gives no clone designation (A31718-2 catalog and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15370 (ELOB_HUMAN, Elongin-B).
- Human Protein Atlas. ELOB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ELOB subcellular location (ICC-IF): Highest expression in MCF-7: 774.8 nTPM.
- Human Protein Atlas. ELOB antibody validation summary (1 antibodies).
- Elongin B promotes breast cancer progression by ubiquitinating tumor suppressor p14/ARF. Cell biology and toxicology 2024 — PMC11039524.
- PubMed PMID:7638163 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ELOB (Elongin-B). Expected localisation Nuclear and cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A31718-2). Validated IHC antibody: A31718-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Cerebellum. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse pancreas tissue; fixative not specified (datasheet A31718-2); 2. Fixation: Image fixative and duration unreported (datasheet A31718-2); 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 A31718-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A31718-2); 8. Primary antibody: Rabbit anti-ELOB, 2-5 μg/ml (datasheet A31718-2); 9. Primary incubation: Overnight at 4 °C (datasheet A31718-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Tissue staining has medium RNA concordance (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unreported (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q15370, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in MCF-7: 774.8 nTPM, Datasheet A31718-2, PMC11039524. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ELOB/immunohistochemistry-ihc-guide.